Patent application title: FAMILY OF PESTICIDAL PROTEINS AND METHODS FOR THEIR USE
Inventors:
Nadine Carozzi (Raleigh, NC, US)
Michael G. Koziel (Raleigh, NC, US)
Nicholas Duck (Apex, NC, US)
Nalini M. Desai (Chapel Hill, NC, US)
Rong Guo (Cary, NC, US)
Rong Guo (Cary, NC, US)
Daniel John Tomso (Bahama, NC, US)
Daniel John Tomso (Bahama, NC, US)
Rebekah Deter (Durham, NC, US)
Tracy Hargiss (Chapel Hill, NC, US)
Tracy Hargiss (Chapel Hill, NC, US)
Assignees:
ATHENIX CORPORATION
IPC8 Class: AC12N1582FI
USPC Class:
800279
Class name: Multicellular living organisms and unmodified parts thereof and related processes method of introducing a polynucleotide molecule into or rearrangement of genetic material within a plant or plant part the polynucleotide confers pathogen or pest resistance
Publication date: 2014-11-20
Patent application number: 20140344999
Abstract:
Compositions and methods for conferring pesticidal activity to bacteria,
plants, plant cells, tissues and seeds are provided. Compositions
comprising a coding sequence for pesticidal polypeptides are provided.
The coding sequences can be used in DNA constructs or expression
cassettes for transformation and expression in plants and bacteria.
Compositions also comprise transformed bacteria, plants, plant cells,
tissues, and seeds. In particular, isolated pesticidal nucleic acid
molecules are provided. Additionally, amino acid sequences corresponding
to the polynucleotides are encompassed. In particular, the present
invention provides for isolated nucleic acid molecules comprising
nucleotide sequences encoding the amino acid sequence shown in SEQ ID NO:
2, 4, 6, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37 or
61, the nucleotide sequence set forth in SEQ ID NO: 1, 3, 5, 8, 10, 12,
14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, or 60, or the nucleotide
sequence deposited in a bacterial host as Accession No. B-30955, B-30956,
B-30957, B-30958, B-30942, B-30939, B-30941, B-50047, B-30959, B-30960,
B-30943, or B-50048, as well as variants and fragments thereof.Claims:
1. An isolated or recombinant nucleic acid molecule comprising a
nucleotide sequence selected from the group consisting of: a) the
nucleotide sequence of SEQ ID NO:18, 3, 5, 8, 10, 12, 14, 16, 20, 22, 24,
26, 28, 30, 32, 34, or 36, or a complement thereof; b) a nucleotide
sequence that encodes a polypeptide comprising the amino acid sequence of
SEQ ID NO:19, 4, 6, 7, 9, 11, 13, 15, 17, 21, 23, 25, 27, 29, 31, 33, 35,
or 37; c) a nucleotide sequence that encodes a polypeptide comprising an
amino acid sequence having at least 95% sequence identity to the amino
acid sequence of SEQ ID NO:19, 4, 6, 7, 9, 11, 13, 15, 17, 21, 23, 25,
27, 29, 31, 33, 35, or 37; and d) the nucleotide sequence of the DNA
insert of the plasmid deposited as Accession No. B-30955, B-30956,
B-30957, B-30958, B-30942, B-30939, B-30941, B-50047, B-30959, B-30960,
B-30943, or B-50048, or a complement thereof.
2. The isolated or recombinant nucleic acid molecule of claim 1, wherein said nucleotide sequence is operably linked to a promoter capable of directing expression of the nucleotide sequence in a plant cell.
3. The isolated or recombinant nucleic acid molecule of claim 1, wherein said nucleotide sequence is a synthetic sequence that has been designed for expression in a plant.
4. A vector comprising the nucleic acid molecule of claim 1.
5. The vector of claim 4, further comprising a nucleic acid molecule encoding a heterologous polypeptide.
6. A host cell that contains the recombinant nucleic acid of claim 1.
7. The host cell of claim 6 that is a bacterial host cell.
8. The host cell of claim 6 that is a plant cell.
9. A transgenic plant comprising the host cell of claim 8.
10. The transgenic plant of claim 9, wherein said plant is selected from the group consisting of maize, sorghum, wheat, cabbage, sunflower, tomato, crucifers, peppers, potato, cotton, rice, soybean, sugarbeet, sugarcane, tobacco, barley, and oilseed rape.
11. A transgenic seed comprising the nucleic acid molecule of claim 1.
12. An isolated polypeptide with pesticidal activity, selected from the group consisting of: a) a polypeptide comprising the amino acid sequence of SEQ ID NO:19, 4, 6, 7, 9, 11, 13, 15, 17, 21, 23, 25, 27, 29, 31, 33, 35, or 37; b) a polypeptide comprising an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 19, 4, 6, 7, 9, 11, 13, 15, 17, 21, 23, 25, 27, 29, 31, 33, 35, or 37; c) a polypeptide that is encoded by the nucleotide sequence of SEQ ID NO:18, 3, 5, 8, 10, 12, 14, 16, 20, 22, 24, 26, 28, 30, 32, 34, or 36; d) a polypeptide encoded by the nucleotide sequence of the DNA insert of the plasmid deposited as Accession No. B-30955, B-30956, B-30957, B-30958, B-30942, B-30939, B-30941, B-50047, B-30959, B-30960, B-30943, or B-50048.
13. The polypeptide of claim 12 further comprising heterologous amino acid sequences.
14. A composition comprising the polypeptide of claim 12.
15. The composition of claim 14, wherein said composition is selected from the group consisting of a powder, dust, pellet, granule, spray, emulsion, colloid, and solution.
16. The composition of claim 14, wherein said composition is prepared by desiccation, lyophilization, homogenization, extraction, filtration, centrifugation, sedimentation, or concentration of a culture of bacterial cells.
17. The composition of claim 14, comprising from about 1% to about 99% by weight of said polypeptide.
18. A method for killing or controlling a lepidopteran, coleopteran, nematode, or dipteran pest population comprising contacting said population with a pesticidally-effective amount of a polypeptide of claim 12.
19. A method for producing a polypeptide with pesticidal activity, comprising culturing the host cell of claim 6 under conditions in which the nucleic acid molecule encoding the polypeptide is expressed.
20. A plant having stably incorporated into its genome a DNA construct comprising a nucleotide sequence that encodes a protein having pesticidal activity, wherein said nucleotide sequence is selected from the group consisting of: a) the nucleotide sequence of SEQ ID NO:18, 3, 5, 8, 10, 12, 14, 16, 20, 22, 24, 26, 28, 30, 32, 34, or 36; b) a nucleotide sequence that encodes a polypeptide comprising the amino acid sequence of SEQ ID NO:19, 4, 6, 7, 9, 11, 13, 15, 17, 21, 23, 25, 27, 29, 31, 33, 35, or 37; c) a nucleotide sequence that encodes a polypeptide comprising an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:19, 4, 6, 7, 9, 11, 13, 15, 17, 21, 23, 25, 27, 29, 31, 33, 35, or 37; and d) the nucleotide sequence of the DNA insert of the plasmid deposited as Accession No. B-30955, B-30956, B-30957, B-30958, B-30942, B-30939, B-30941, B-50047, B-30959, B-30960, B-30943, or B-50048; wherein said nucleotide sequence is operably linked to a promoter that drives expression of a coding sequence in a plant cell.
21. The plant of claim 20, wherein said plant is a plant cell.
22. A method for protecting a plant from a pest, comprising introducing into said plant or cell thereof at least one expression vector comprising a nucleotide sequence that encodes a pesticidal polypeptide, wherein said nucleotide sequence is selected from the group consisting of: a) the nucleotide sequence of SEQ ID NO:18, 3, 5, 8, 10, 12, 14, 16, 20, 22, 24, 26, 28, 30, 32, 34, or 36; b) a nucleotide sequence that encodes a polypeptide comprising the amino acid sequence of SEQ ID NO:19, 4, 6, 7, 9, 11, 13, 15, 17, 21, 23, 25, 27, 29, 31, 33, 35, or 37; c) a nucleotide sequence that encodes a polypeptide comprising an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:19, 4, 6, 7, 9, 11, 13, 15, 17, 21, 23, 25, 27, 29, 31, 33, 35, or 37; and d) the nucleotide sequence of the DNA insert of the plasmid deposited as Accession No. B-30955, B-30956, B-30957, B-30958, B-30942, B-30939, B-30941, B-50047, B-30959, B-30960, B-30943, or B-50048.
23. The method of claim 22, wherein said plant produces a pesticidal polypeptide having pesticidal activity against a lepidopteran, coleopteran, nematode, or dipteran pest.
Description:
CROSS REFERENCE TO RELATED APPLICATION
[0001] This application is a divisional of U.S. patent application Ser. No. 13/168,454, filed Jun. 24, 2011, which is a continuation of U.S. patent application Ser. No. 11/763,947, filed Jun. 15, 2007, which claims the benefit of U.S. Provisional Application Ser. Nos. 60/814,007, filed Jun. 15, 2006; 60/813,859, filed Jun. 15, 2006; 60/814,420, filed Jun. 16, 2006; 60/814,212, filed Jun. 16, 2006; and 60/814,989, filed Jun. 20, 2006, the contents of which are herein incorporated by reference in their entirety.
REFERENCE TO SEQUENCE LISTING SUBMITTED ELECTRONICALLY
[0002] The official copy of the sequence listing is submitted electronically via EFS-Web as an ASCII formatted sequence listing with a file named "APA039US01NDSEQLIST.txt", created on Aug. 5, 2014, and having a size of 305 kilobytes and is filed concurrently with the specification. The sequence listing contained in this ASCII formatted document is part of the specification and is herein incorporated by reference in its entirety.
FIELD OF THE INVENTION
[0003] This invention relates to the field of molecular biology. Provided are novel genes that encode pesticidal proteins. These proteins and the nucleic acid sequences that encode them are useful in preparing pesticidal formulations and in the production of transgenic pest-resistant plants.
BACKGROUND OF THE INVENTION
[0004] Bacillus thuringiensis is a Gram-positive spore forming soil bacterium characterized by its ability to produce crystalline inclusions that are specifically toxic to certain orders and species of insects, but are harmless to plants and other non-targeted organisms. For this reason, compositions including Bacillus thuringiensis strains or their insecticidal proteins can be used as environmentally-acceptable insecticides to control agricultural insect pests or insect vectors for a variety of human or animal diseases.
[0005] Crystal (Cry) proteins (delta-endotoxins) from Bacillus thuringiensis have potent insecticidal activity against predominantly Lepidopteran, Dipteran, and Coleopteran larvae. These proteins also have shown activity against Hymenoptera, Homoptera, Phthiraptera, Mallophaga, and Acari pest orders, as well as other invertebrate orders such as Nemathelminthes, Platyhelminthes, and Sarcomastigorphora (Feitelson (1993) The Bacillus Thuringiensis family tree. In Advanced Engineered Pesticides, Marcel Dekker, Inc., New York, N.Y.) These proteins were originally classified as CryI to CryV based primarily on their insecticidal activity. The major classes were Lepidoptera-specific (I), Lepidoptera- and Diptera-specific (II), Coleoptera-specific (III), Diptera-specific (IV), and nematode-specific (V) and (VI). The proteins were further classified into subfamilies; more highly related proteins within each family were assigned divisional letters such as Cry1A, Cry1B, Cry1C, etc. Even more closely related proteins within each division were given names such as Cry1C1, Cry1C2, etc.
[0006] A new nomenclature was recently described for the Cry genes based upon amino acid sequence homology rather than insect target specificity (Crickmore et al. (1998) Microbiol. Mol. Biol. Rev. 62:807-813). In the new classification, each toxin is assigned a unique name incorporating a primary rank (an Arabic number), a secondary rank (an uppercase letter), a tertiary rank (a lowercase letter), and a quaternary rank (another Arabic number). In the new classification, Roman numerals have been exchanged for Arabic numerals in the primary rank. Proteins with less than 45% sequence identity have different primary ranks, and the criteria for secondary and tertiary ranks are 78% and 95%, respectively.
[0007] The crystal protein does not exhibit insecticidal activity until it has been ingested and solubilized in the insect midgut. The ingested protoxin is hydrolyzed by proteases in the insect digestive tract to an active toxic molecule. (Hofte and Whiteley (1989) Microbiol. Rev. 53:242-255). This toxin binds to apical brush border receptors in the midgut of the target larvae and inserts into the apical membrane creating ion channels or pores, resulting in larval death.
[0008] Delta-endotoxins generally have five conserved sequence domains, and three conserved structural domains (see, for example, de Maagd et al. (2001) Trends Genetics 17:193-199). The first conserved structural domain consists of seven alpha helices and is involved in membrane insertion and pore formation. Domain II consists of three beta-sheets arranged in a Greek key configuration, and domain III consists of two antiparallel beta-sheets in "jelly-roll" formation (de Maagd et al., 2001, supra). Domains II and III are involved in receptor recognition and binding, and are therefore considered determinants of toxin specificity.
[0009] Because of the devastation that insects can confer, and the improvement in yield by controlling insect pests, there is a continual need to discover new forms of pesticidal toxins.
SUMMARY OF INVENTION
[0010] Compositions and methods for conferring pesticidal activity to bacteria, plants, plant cells, tissues and seeds are provided. Compositions include nucleic acid molecules encoding sequences for pesticidal and insectidal polypeptides, vectors comprising those nucleic acid molecules, and host cells comprising the vectors. Compositions also include the pesticidal polypeptide sequences and antibodies to those polypeptides. The nucleotide sequences can be used in DNA constructs or expression cassettes for transformation and expression in organisms, including microorganisms and plants. The nucleotide or amino acid sequences may be synthetic sequences that have been designed for expression in an organism including, but not limited to, a microorganism or a plant. Compositions also comprise transformed bacteria, plants, plant cells, tissues, and seeds.
[0011] In particular, isolated nucleic acid molecules are provided that encode a pesticidal protein. Additionally, amino acid sequences corresponding to the pesticidal protein are encompassed. In particular, the present invention provides for an isolated nucleic acid molecule comprising a nucleotide sequence encoding the amino acid sequence shown in SEQ ID NO:2, 4, 6, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37 or 61, a nucleotide sequence set forth in SEQ ID NO:1, 3, 5, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, or 60, or the delta-endotoxin nucleotide sequence of the DNA insert of the plasmid deposited in a bacterial host as Accession No. B-30955, B-30956, B-30957, B-30958, B-30942, B-30939, B-30941, B-50047, B-30959, B-30960, B-30943, or B-50048, as well as variants and fragments thereof. Nucleotide sequences that are complementary to a nucleotide sequence of the invention, or that hybridize to a sequence of the invention are also encompassed.
[0012] Methods are provided for producing the polypeptides of the invention, and for using those polypeptides for controlling or killing a lepidopteran, coleopteran, nematode, or dipteran pest. Methods and kits for detecting the nucleic acids and polypeptides of the invention in a sample are also included.
[0013] The compositions and methods of the invention are useful for the production of organisms with enhanced pest resistance or tolerance. These organisms and compositions comprising the organisms are desirable for agricultural purposes. The compositions of the invention are also useful for generating altered or improved proteins that have pesticidal activity, or for detecting the presence of pesticidal proteins or nucleic acids in products or organisms.
DESCRIPTION OF FIGURES
[0014] FIG. 1 shows an alignment of AXMI-022 with the Iota1b from Clostridium perfringens (SEQ ID NO:49), Isp1A from Brevibacillus laterosporus (SEQ ID NO:50), Isp1B from Brevibacillus laterosporus (SEQ ID NO:51), Vip1Ab from Bacillus thuringiensis (SEQ ID NO:52), and Vip1Ac from Bacillus thuringiensis (SEQ ID NO:53).
[0015] FIG. 2 shows an alignment of AXMI-022 with Vip1Ab (SEQ ID NO:52).
[0016] FIG. 3 shows an alignment of AXMI-022 with Cry 37Aa1 from Bacillus thuringiensis (SEQ ID NO:54).
[0017] FIG. 4 shows an alignment of AXMI-023 with the Vip2 pesticidal protein (SEQ ID NO:55), Isp2a from Brevibacillus laterosporus (SEQ ID NO:56) and Iota toxin component Ia from Clostridium perfringens (SEQ ID NO:57).
DETAILED DESCRIPTION
[0018] The present invention is drawn to compositions and methods for regulating pest resistance or tolerance in organisms, particularly plants or plant cells. By "resistance" is intended that the pest (e.g., insect) is killed upon ingestion or other contact with the polypeptides of the invention. By "tolerance" is intended an impairment or reduction in the movement, feeding, reproduction, or other functions of the pest. The methods involve transforming organisms with a nucleotide sequence encoding a pesticidal protein of the invention. In particular, the nucleotide sequences of the invention are useful for preparing plants and microorganisms that possess pesticidal activity. Thus, transformed bacteria, plants, plant cells, plant tissues and seeds are provided. Compositions are pesticidal nucleic acids and proteins of Bacillus or other species. The sequences find use in the construction of expression vectors for subsequent transformation into organisms of interest, as probes for the isolation of other homologous (or partially homologous) genes, and for the generation of altered pesticidal proteins by methods known in the art, such as domain swapping or DNA shuffling. The proteins find use in controlling or killing lepidopteran, coleopteran, dipteran, and nematode pest populations and for producing compositions with pesticidal activity.
[0019] Plasmids containing the nucleotide sequences of the invention were deposited in the permanent collection of the Agricultural Research Service Culture Collection, Northern Regional Research Laboratory (NRRL), 1815 North University Street, Peoria, Ill. 61604, United States of America, in accordance with Table 1. This deposit will be maintained under the terms of the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure. Access to these deposits will be available during the pendency of the application to the Commissioner of Patents and Trademarks and persons determined by the Commissioner to be entitled thereto upon request. Upon allowance of any claims in the application, the Applicants will make available to the public, pursuant to 37 C.F.R. §1.808, sample(s) of the deposit with the ATCC. This deposit was made merely as a convenience for those of skill in the art and is not an admission that a deposit is required under 35 U.S.C. §112.
TABLE-US-00001 TABLE 1 Microorganism Deposit NRRL Gene Strain Clone number Deposit Date axmi-011 ATX13008 pAX4600 B-30961 Jul. 21, 2006 axmi-012 ATX13026 pAX012 B-30955 Jul. 21, 2006 axmi-013 ATX13002 pAX013 B-30956 Jul. 21, 2006 axmi-015 ATX13026 pAX015 B-30957 Jul. 21, 2006 axmi-019 ATX14875 pAX019 B-30958 Jul. 21, 2006 axmi-044 ATX14759 pAX2599 B-30942 Jun. 15, 2006 axmi-037 ATX1489 pAX2558 B-30939 Jun. 15, 2006 axmi-043 ATX15398 pAX2597 B-30941 Jun. 15, 2006 axmi-033 ATX14833 pAX4341 B-50047 May 29, 2007 axmi-034 ATX14833 pAX4341 B-50047 May 29, 2007 axmi-022 ATX13045 pAX022 B-30959 Jul. 21, 2006 axmi-023 ATX13045 pAX023 B-30960 Jul. 21, 2006 axmi-041 ATX21738 pAX4310 B-30943 Jun. 15, 2006 axmi-063 ATX12972 pAX5036 B-50048 May 29, 2007 axmi-064 ATX12972 pAX5036 B-50048 May 29, 2007
[0020] By "pesticidal toxin" or "pesticidal protein" is intended a toxin that has toxic activity against one or more pests, including, but not limited to, members of the Lepidoptera, Diptera, and Coleoptera orders, or the Nematoda phylum, or a protein that has homology to such a protein. Pesticidal proteins have been isolated from organisms including, for example, Bacillus sp., Clostridium bifermentans and Paenibacillus popilliae. Pesticidal proteins include amino acid sequences deduced from the full-length nucleotide sequences disclosed herein, and amino acid sequences that are shorter than the full-length sequences, either due to the use of an alternate downstream start site, or due to processing that produces a shorter protein having pesticidal activity. Processing may occur in the organism the protein is expressed in, or in the pest after ingestion of the protein.
[0021] Pesticidal proteins encompass delta-endotoxins. Delta-endotoxins include proteins identified as cry1 through cry43, cyt1 and cyt2, and Cyt-like toxin. There are currently over 250 known species of delta-endotoxins with a wide range of specificities and toxicities. For an expansive list see Crickmore et al. (1998), Microbiol. Mol. Biol. Rev. 62:807-813, and for regular updates see Crickmore et al. (2003) "Bacillus thuringiensis toxin nomenclature," at www.biols.susx.ac.uk/Home/Neil_Crickmore/Bt/index.
[0022] Also provided herein are nucleotide sequences encoding polypeptides with homology to several other known classes of pesticidal protein toxins. For example, axmi-011, axmi-012, axmi-015, axmi-032, axmi-044, axmi-033, axmi-034, axmi-022, axmi-063, and axmi-064 demonstrate homology to pesticidal binary toxins such as VIP, Bin, and MTX toxins. The VIP1/VIP2 toxins (see, for example, U.S. Pat. No. 5,770,696, herein incorporated by reference in its entirety) are binary pesticidal toxins that exhibit strong activity on insects by a mechanism believed to involve receptor-mediated endocytosis followed by cellular toxification, similar to the mode of action of other binary ("A/B") toxins. A/B toxins such as VIP, C2, CDT, CST, or the B. anthracis edema and lethal toxins initially interact with target cells via a specific, receptor-mediated binding of "B" components as monomers. These monomers then form homoheptamers. The "B" heptamer-receptor complex then acts as a docking platform that subsequently binds and allows the translocation of an enzymatic "A" component(s) into the cytosol via receptor-mediated endocytosis. Once inside the cell's cytosol, "A" components inhibit normal cell function by, for example, ADP-ribosylation of G-actin, or increasing intracellular levels of cyclic AMP (cAMP). See Barth et al. (2004) Microbiol Mol Biol Rev 68:373-402, herein incorporated by reference in its entirety.
[0023] Aside from the A/B type binary toxins, other types of binary toxins that act as pesticidal proteins are known in the art. Cry34Ab1 and Cry35Ab1 comprise a binary toxin with pesticidal activity that was identified from strain PS149B1 (Ellis et al. (2002) Appl Environ Microbiol. 68:1137-45, herein incorporated by reference in its entirety). These toxins have molecular masses of approximately 14 and 44 kDa, respectively. Other binary toxins with similar organization and homology to Cry34Aa and Cry34Ab have been identified (Baum et al. (2004) Appl Environ Microbiol. 70:4889-98, herein incorporated by reference in its entirety).
[0024] BinA and BinB are proteins from Bacillus sphaericus that comprise a mosquitocidal binary toxin protein (Baumann et al. (1991) Micriobiol. Rev. 55:425-36). Cry35 exhibits amino acid similarity to these BinA and BinB proteins. Cry36 (ET69) and Cry38 (ET75) (International Patent Application No. WO/00/66742-B, herein incorporated by reference in its entirety) are independently isolated peptides that also exhibit amino acid similarity to BinA and BinB, and thus are likely to comprise binary toxins.
[0025] Cry23 (also known as cryET33; U.S. Pat. No. 6,063,756, herein incorporated by reference in its entirety) and Cry37 (also known as cryET34; U.S. Pat. No. 6,063,756, herein incorporated by reference in its entirety) also appear to be binary pesticidal toxins. Cry23 also exhibits homology to MTX2 and MTX3 toxins. The term "MTX" is used in the art to delineate a set of pesticidal proteins that are produced by Bacillus sphaericus. The first of these, often referred to in the art as MTX1, is synthesized as a parasporal crystal which is toxic to mosquitoes. The major components of the crystal are two proteins of 51 and 42 kDa, Since the presence of both proteins are required for toxicity, MTX1 is considered a "binary" toxin (Baumann et al. (1991) Microbiol. Rev. 55:425-436).
[0026] By analysis of different Bacillus sphaericus strains with differing toxicities, two new classes of MTX toxins have been identified. MTX2 and MTX3 represent separate, related classes of pesticidal toxins that exhibit pesticidal activity. See, for example, Baumann et al. (1991) Microbiol. Rev. 55:425-436, herein incorporated by reference in its entirety. MTX2 is a 100-kDa toxin. More recently MTX3 has been identified as a separate toxin, though the amino acid sequence of MTX3 from B. sphaericus is 38% identitical to the MTX2 toxin of B. sphaericus SSII-1 (Liu, et al. (1996) Appl. Environ. Microbiol. 62: 2174-2176).
[0027] Thus, provided herein are novel isolated nucleotide sequences that confer pesticidal activity. These isolated nucleotide sequences encode polypeptides with homology to known delta-endotoxins or binary toxins. Also provided are the amino acid sequences of the pesticidal proteins. The protein resulting from translation of this gene allows cells to control or kill pests that ingest it.
Isolated Nucleic Acid Molecules, and Variants and Fragments Thereof
[0028] One aspect of the invention pertains to isolated or recombinant nucleic acid molecules comprising nucleotide sequences encoding pesticidal proteins and polypeptides or biologically active portions thereof, as well as nucleic acid molecules sufficient for use as hybridization probes to identify nucleic acid molecules encoding proteins with regions of sequence homology. As used herein, the term "nucleic acid molecule" is intended to include DNA molecules (e.g., recombinant DNA, cDNA or genomic DNA) and RNA molecules (e.g., mRNA) and analogs of the DNA or RNA generated using nucleotide analogs. The nucleic acid molecule can be single-stranded or double-stranded, but preferably is double-stranded DNA.
[0029] An "isolated" or "purified" nucleic acid molecule or protein, or biologically active portion thereof, is substantially free of other cellular material, or culture medium when produced by recombinant techniques, or substantially free of chemical precursors or other chemicals when chemically synthesized. Preferably, an "isolated" nucleic acid is free of sequences (preferably protein encoding sequences) that naturally flank the nucleic acid (i.e., sequences located at the 5' and 3' ends of the nucleic acid) in the genomic DNA of the organism from which the nucleic acid is derived. For purposes of the invention, "isolated" when used to refer to nucleic acid molecules excludes isolated chromosomes. For example, in various embodiments, the isolated nucleic acid molecule encoding a pesticidal protein can contain less than about 5 kb, 4 kb, 3 kb, 2 kb, 1 kb, 0.5 kb, or 0.1 kb of nucleotide sequences that naturally flank the nucleic acid molecule in genomic DNA of the cell from which the nucleic acid is derived. A pesticidal protein that is substantially free of cellular material includes preparations of protein having less than about 30%, 20%, 10%, or 5% (by dry weight) of non-pesticidal protein (also referred to herein as a "contaminating protein").
[0030] Nucleotide sequences encoding the proteins of the present invention include the sequence set forth in SEQ ID NO:1, 3, 5, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, or 60, or the nucleotide sequence deposited in a bacterial host as Accession No. B-30955, B-30956, B-30957, B-30958, B-30942, B-30939, B-30941, B-50047, B-30959, B-30960, B-30943, or B-50048, and variants, fragments, and complements thereof. By "complement" is intended a nucleotide sequence that is sufficiently complementary to a given nucleotide sequence such that it can hybridize to the given nucleotide sequence to thereby form a stable duplex. The corresponding amino acid sequence for the pesticidal protein encoded by this nucleotide sequence are set forth in SEQ ID NO:2, 4, 6, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37 or 61.
[0031] Nucleic acid molecules that are fragments of these nucleotide sequences encoding pesticidal proteins are also encompassed by the present invention. By "fragment" is intended a portion of the nucleotide sequence encoding a pesticidal protein. A fragment of a nucleotide sequence may encode a biologically active portion of a pesticidal protein, or it may be a fragment that can be used as a hybridization probe or PCR primer using methods disclosed below. Nucleic acid molecules that are fragments of a nucleotide sequence encoding a pesticidal protein comprise at least about 50, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1350, 1400 contiguous nucleotides, or up to the number of nucleotides present in a full-length nucleotide sequence encoding a pesticidal protein disclosed herein (for example, 957 nucleotides for SEQ ID NO: 1; 927 nucleotides for SEQ ID NO:3, 1017 nucleotides for SEQ ID NO:5; 1422 nucleotides for SEQ ID NO:8, 1053 nucleotides for SEQ ID NO:10; 1062 nucleotides for SEQ ID NO: 12, 942 nucleotides for SEQ ID NO: 14, etc.) depending upon the intended use. By "contiguous" nucleotides is intended nucleotide residues that are immediately adjacent to one another. Fragments of the nucleotide sequences of the present invention will encode protein fragments that retain the biological activity of the pesticidal protein and, hence, retain pesticidal activity. By "retains activity" is intended that the fragment will have at least about 30%, at least about 50%, at least about 70%, 80%, 90%, 95% or higher of the pesticidal activity of the pesticidal protein. Methods for measuring pesticidal activity are well known in the art. See, for example, Czapla and Lang (1990) J. Econ. Entomol. 83:2480-2485; Andrews et al. (1988) Biochem. J. 252:199-206; Marrone et al. (1985) J. of Economic Entomology 78:290-293; and U.S. Pat. No. 5,743,477, all of which are herein incorporated by reference in their entirety.
[0032] A fragment of a nucleotide sequence encoding a pesticidal protein that encodes a biologically active portion of a protein of the invention will encode at least about 15, 25, 30, 50, 75, 100, 125, 150, 175, 200, 250, 300, 350, 400, 450 contiguous amino acids, or up to the total number of amino acids present in a full-length pesticidal protein of the invention (for example, 318 amino acids for SEQ ID NO:2, 308 amino acids for SEQ ID NO:4, 338 amino acids for SEQ ID NO:6, 296 amino acids for SEQ ID NO:7, 473 amino acids for SEQ ID NO:9, 351 amino acids for SEQ ID NO:11, 353 amino acids for SEQ ID NO:13, and 314 amino acids for SEQ ID NO:15, etc.).
[0033] Preferred pesticidal proteins of the present invention are encoded by a nucleotide sequence sufficiently identical to the nucleotide sequence of SEQ ID NO: 1, 3, 5, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, or 60. By "sufficiently identical" is intended an amino acid or nucleotide sequence that has at least about 60% or 65% sequence identity, about 70% or 75% sequence identity, about 80% or 85% sequence identity, about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or greater sequence identity compared to a reference sequence using one of the alignment programs described herein using standard parameters. One of skill in the art will recognize that these values can be appropriately adjusted to determine corresponding identity of proteins encoded by two nucleotide sequences by taking into account codon degeneracy, amino acid similarity, reading frame positioning, and the like.
[0034] To determine the percent identity of two amino acid sequences or of two nucleic acids, the sequences are aligned for optimal comparison purposes. The percent identity between the two sequences is a function of the number of identical positions shared by the sequences (i.e., percent identity=number of identical positions/total number of positions (e.g., overlapping positions)×100). In one embodiment, the two sequences are the same length. The percent identity between two sequences can be determined using techniques similar to those described below, with or without allowing gaps. In calculating percent identity, typically exact matches are counted.
[0035] The determination of percent identity between two sequences can be accomplished using a mathematical algorithm. A nonlimiting example of a mathematical algorithm utilized for the comparison of two sequences is the algorithm of Karlin and Altschul (1990) Proc. Natl. Acad. Sci. USA 87:2264, modified as in Karlin and Altschul (1993) Proc. Natl. Acad. Sci. USA 90:5873-5877. Such an algorithm is incorporated into the BLASTN and BLASTX programs of Altschul et al. (1990) J. Mol. Biol. 215:403. BLAST nucleotide searches can be performed with the BLASTN program, score=100, wordlength=12, to obtain nucleotide sequences homologous to pesticidal-like nucleic acid molecules of the invention. BLAST protein searches can be performed with the BLASTX program, score=50, wordlength=3, to obtain amino acid sequences homologous to pesticidal protein molecules of the invention. To obtain gapped alignments for comparison purposes, Gapped BLAST (in BLAST 2.0) can be utilized as described in Altschul et al. (1997) Nucleic Acids Res. 25:3389. Alternatively, PSI-Blast can be used to perform an iterated search that detects distant relationships between molecules. See Altschul et al. (1997) supra. When utilizing BLAST, Gapped BLAST, and PSI-Blast programs, the default parameters of the respective programs (e.g., BLASTX and BLASTN) can be used. Alignment may also be performed manually by inspection.
[0036] Another non-limiting example of a mathematical algorithm utilized for the comparison of sequences is the ClustalW algorithm (Higgins et al. (1994) Nucleic Acids Res. 22:4673-4680). ClustalW compares sequences and aligns the entirety of the amino acid or DNA sequence, and thus can provide data about the sequence conservation of the entire amino acid sequence. The ClustalW algorithm is used in several commercially available DNA/amino acid analysis software packages, such as the ALIGNX module of the Vector NTI Program Suite (Invitrogen Corporation, Carlsbad, Calif.). After alignment of amino acid sequences with ClustalW, the percent amino acid identity can be assessed. A non-limiting example of a software program useful for analysis of ClustalW alignments is GENEDOC®. GENEDOC® (Karl Nicholas) allows assessment of amino acid (or DNA) similarity and identity between multiple proteins. Another non-limiting example of a mathematical algorithm utilized for the comparison of sequences is the algorithm of Myers and Miller (1988) CABIOS 4:11-17. Such an algorithm is incorporated into the ALIGN program (version 2.0), which is part of the GCG Wisconsin Genetics Software Package, Version 10 (available from Accelrys, Inc., 9685 Scranton Rd., San Diego, Calif., USA). When utilizing the ALIGN program for comparing amino acid sequences, a PAM 120 weight residue table, a gap length penalty of 12, and a gap penalty of 4 can be used.
[0037] Unless otherwise stated, GAP Version 10, which uses the algorithm of Needleman and Wunsch (1970) J. Mol. Biol. 48 (3):443-453, will be used to determine sequence identity or similarity using the following parameters: % identity and % similarity for a nucleotide sequence using GAP Weight of 50 and Length Weight of 3, and the nwsgapdna.cmp scoring matrix; % identity or % similarity for an amino acid sequence using GAP weight of 8 and length weight of 2, and the BLOSUM62 scoring program. Equivalent programs may also be used. By "equivalent program" is intended any sequence comparison program that, for any two sequences in question, generates an alignment having identical nucleotide residue matches and an identical percent sequence identity when compared to the corresponding alignment generated by GAP Version 10.
[0038] The invention also encompasses variant nucleic acid molecules. "Variants" of the pesticidal protein encoding nucleotide sequences include those sequences that encode the pesticidal proteins disclosed herein but that differ conservatively because of the degeneracy of the genetic code as well as those that are sufficiently identical as discussed above. Naturally occurring allelic variants can be identified with the use of well-known molecular biology techniques, such as polymerase chain reaction (PCR) and hybridization techniques as outlined below. Variant nucleotide sequences also include synthetically derived nucleotide sequences that have been generated, for example, by using site-directed mutagenesis but which still encode the pesticidal proteins disclosed in the present invention as discussed below. Variant proteins encompassed by the present invention are biologically active, that is they continue to possess the desired biological activity of the native protein, that is, pesticidal activity. By "retains activity" is intended that the variant will have at least about 30%, at least about 50%, at least about 70%, or at least about 80% of the pesticidal activity of the native protein. Methods for measuring pesticidal activity are well known in the art. See, for example, Czapla and Lang (1990) J. Econ. Entomol. 83: 2480-2485; Andrews et al. (1988) Biochem. J. 252:199-206; Marrone et al. (1985) J. of Economic Entomology 78:290-293; and U.S. Pat. No. 5,743,477, all of which are herein incorporated by reference in their entirety.
[0039] The skilled artisan will further appreciate that changes can be introduced by mutation of the nucleotide sequences of the invention thereby leading to changes in the amino acid sequence of the encoded pesticidal proteins, without altering the biological activity of the proteins. Thus, variant isolated nucleic acid molecules can be created by introducing one or more nucleotide substitutions, additions, or deletions into the corresponding nucleotide sequence disclosed herein, such that one or more amino acid substitutions, additions or deletions are introduced into the encoded protein. Mutations can be introduced by standard techniques, such as site-directed mutagenesis and PCR-mediated mutagenesis. Such variant nucleotide sequences are also encompassed by the present invention.
[0040] For example, conservative amino acid substitutions may be made at one or more, predicted, nonessential amino acid residues. A "nonessential" amino acid residue is a residue that can be altered from the wild-type sequence of a pesticidal protein without altering the biological activity, whereas an "essential" amino acid residue is required for biological activity. A "conservative amino acid substitution" is one in which the amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine).
[0041] Delta-endotoxins generally have five conserved sequence domains, and three conserved structural domains (see, for example, de Maagd et al. (2001) Trends Genetics 17:193-199). The first conserved structural domain consists of seven alpha helices and is involved in membrane insertion and pore formation. Domain II consists of three beta-sheets arranged in a Greek key configuration, and domain III consists of two antiparallel beta-sheets in "jelly-roll" formation (de Maagd et al., 2001, supra). Domains II and III are involved in receptor recognition and binding, and are therefore considered determinants of toxin specificity.
[0042] Amino acid substitutions may be made in nonconserved regions that retain function. In general, such substitutions would not be made for conserved amino acid residues, or for amino acid residues residing within a conserved motif, where such residues are essential for protein activity. Examples of residues that are conserved and that may be essential for protein activity include, for example, residues that are identical between all proteins contained in an alignment of similar or related toxins to the sequences of the invention (e.g., residues that are identical between all proteins contained in the alignment in FIG. 1, 2, 3, or 4). Examples of residues that are conserved but that may allow conservative amino acid substitutions and still retain activity include, for example, residues that have only conservative substitutions between all proteins contained in an alignment of similar or related toxins to the sequences of the invention (e.g., residues that have only conservative substitutions between all proteins contained in the alignment in FIG. 1, 2, 3, or 4). However, one of skill in the art would understand that functional variants may have minor conserved or nonconserved alterations in the conserved residues.
[0043] Alternatively, variant nucleotide sequences can be made by introducing mutations randomly along all or part of the coding sequence, such as by saturation mutagenesis, and the resultant mutants can be screened for ability to confer pesticidal activity to identify mutants that retain activity. Following mutagenesis, the encoded protein can be expressed recombinantly, and the activity of the protein can be determined using standard assay techniques.
[0044] Using methods such as PCR, hybridization, and the like corresponding pesticidal sequences can be identified, such sequences having substantial identity to the sequences of the invention. See, for example, Sambrook and Russell (2001) Molecular Cloning: A Laboratory Manual. (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.) and Innis, et al. (1990) PCR Protocols: A Guide to Methods and Applications (Academic Press, NY).
[0045] In a hybridization method, all or part of the pesticidal nucleotide sequence can be used to screen cDNA or genomic libraries. Methods for construction of such cDNA and genomic libraries are generally known in the art and are disclosed in Sambrook and Russell, 2001, supra. The so-called hybridization probes may be genomic DNA fragments, cDNA fragments, RNA fragments, or other oligonucleotides, and may be labeled with a detectable group such as 32P, or any other detectable marker, such as other radioisotopes, a fluorescent compound, an enzyme, or an enzyme co-factor. Probes for hybridization can be made by labeling synthetic oligonucleotides based on the known pesticidal protein-encoding nucleotide sequence disclosed herein. Degenerate primers designed on the basis of conserved nucleotides or amino acid residues in the nucleotide sequence or encoded amino acid sequence can additionally be used. The probe typically comprises a region of nucleotide sequence that hybridizes under stringent conditions to at least about 12, at least about 25, at least about 50, 75, 100, 125, 150, 175, or 200 consecutive nucleotides of nucleotide sequence encoding a pesticidal protein of the invention or a fragment or variant thereof. Methods for the preparation of probes for hybridization are generally known in the art and are disclosed in Sambrook and Russell, 2001, supra herein incorporated by reference.
[0046] For example, an entire pesticidal protein sequence disclosed herein, or one or more portions thereof, may be used as a probe capable of specifically hybridizing to corresponding pesticidal protein-like sequences and messenger RNAs. To achieve specific hybridization under a variety of conditions, such probes include sequences that are unique and are preferably at least about 10 nucleotides in length, or at least about 20 nucleotides in length. Such probes may be used to amplify corresponding pesticidal sequences from a chosen organism by PCR. This technique may be used to isolate additional coding sequences from a desired organism or as a diagnostic assay to determine the presence of coding sequences in an organism. Hybridization techniques include hybridization screening of plated DNA libraries (either plaques or colonies; see, for example, Sambrook et al. (1989) Molecular Cloning: A Laboratory Manual (2d ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.).
[0047] Hybridization of such sequences may be carried out under stringent conditions. By "stringent conditions" or "stringent hybridization conditions" is intended conditions under which a probe will hybridize to its target sequence to a detectably greater degree than to other sequences (e.g., at least 2-fold over background). Stringent conditions are sequence-dependent and will be different in different circumstances. By controlling the stringency of the hybridization and/or washing conditions, target sequences that are 100% complementary to the probe can be identified (homologous probing). Alternatively, stringency conditions can be adjusted to allow some mismatching in sequences so that lower degrees of similarity are detected (heterologous probing). Generally, a probe is less than about 1000 nucleotides in length, preferably less than 500 nucleotides in length.
[0048] Typically, stringent conditions will be those in which the salt concentration is less than about 1.5 M Na ion, typically about 0.01 to 1.0 M Na ion concentration (or other salts) at pH 7.0 to 8.3 and the temperature is at least about 30° C. for short probes (e.g., 10 to 50 nucleotides) and at least about 60° C. for long probes (e.g., greater than 50 nucleotides). Stringent conditions may also be achieved with the addition of destabilizing agents such as formamide. Exemplary low stringency conditions include hybridization with a buffer solution of 30 to 35% formamide, 1 M NaCl, 1% SDS (sodium dodecyl sulphate) at 37° C., and a wash in 1× to 2×SSC (20×SSC=3.0 M NaCl/0.3 M trisodium citrate) at 50 to 55° C. Exemplary moderate stringency conditions include hybridization in 40 to 45% formamide, 1.0 M NaCl, 1% SDS at 37° C., and a wash in 0.5× to 1×SSC at 55 to 60° C. Exemplary high stringency conditions include hybridization in 50% formamide, 1 M NaCl, 1% SDS at 37° C., and a wash in 0.1×SSC at 60 to 65° C. Optionally, wash buffers may comprise about 0.1% to about 1% SDS. Duration of hybridization is generally less than about 24 hours, usually about 4 to about 12 hours.
[0049] Specificity is typically the function of post-hybridization washes, the critical factors being the ionic strength and temperature of the final wash solution. For DNA-DNA hybrids, the Tm can be approximated from the equation of Meinkoth and Wahl (1984) Anal. Biochem. 138:267-284: Tm=81.5° C.+16.6 (log M)+0.41 (% GC)-0.61 (% form)-500/L; where M is the molarity of monovalent cations, % GC is the percentage of guanosine and cytosine nucleotides in the DNA, % form is the percentage of formamide in the hybridization solution, and L is the length of the hybrid in base pairs. The Tm is the temperature (under defined ionic strength and pH) at which 50% of a complementary target sequence hybridizes to a perfectly matched probe. Tm is reduced by about 1° C. for each 1% of mismatching; thus, Tm, hybridization, and/or wash conditions can be adjusted to hybridize to sequences of the desired identity. For example, if sequences with ≧90% identity are sought, the Tm can be decreased 10° C. Generally, stringent conditions are selected to be about 5° C. lower than the thermal melting point (Tm) for the specific sequence and its complement at a defined ionic strength and pH. However, severely stringent conditions can utilize a hybridization and/or wash at 1, 2, 3, or 4° C. lower than the thermal melting point (Tm); moderately stringent conditions can utilize a hybridization and/or wash at 6, 7, 8, 9, or 10° C. lower than the thermal melting point (Tm); low stringency conditions can utilize a hybridization and/or wash at 11, 12, 13, 14, 15, or 20° C. lower than the thermal melting point (Tm). Using the equation, hybridization and wash compositions, and desired Tm, those of ordinary skill will understand that variations in the stringency of hybridization and/or wash solutions are inherently described. If the desired degree of mismatching results in a Tm of less than 45° C. (aqueous solution) or 32° C. (formamide solution), it is preferred to increase the SSC concentration so that a higher temperature can be used. An extensive guide to the hybridization of nucleic acids is found in Tijssen (1993) Laboratory Techniques in Biochemistry and Molecular Biology--Hybridization with Nucleic Acid Probes, Part I, Chapter 2 (Elsevier, New York); and Ausubel et al., eds. (1995) Current Protocols in Molecular Biology, Chapter 2 (Greene Publishing and Wiley-Interscience, New York). See Sambrook et al. (1989) Molecular Cloning: A Laboratory Manual (2d ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.).
Isolated Proteins and Variants and Fragments Thereof
[0050] Pesticidal proteins are also encompassed within the present invention. By "pesticidal protein" is intended a protein having the amino acid sequence set forth in SEQ ID NO:2, 4, 6, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37 or 61. Fragments, biologically active portions, and variants thereof are also provided, and may be used to practice the methods of the present invention.
[0051] "Fragments" or "biologically active portions" include polypeptide fragments comprising amino acid sequences sufficiently identical to the amino acid sequence set forth in SEQ ID NO:2, 4, 6, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37 or 61, and that exhibit pesticidal activity (for example, SEQ ID NO:7). A biologically active portion of a pesticidal protein can be a polypeptide that is, for example, 10, 25, 50, 100, 150, 200, 250 or more amino acids in length. Such biologically active portions can be prepared by recombinant techniques and evaluated for pesticidal activity. Methods for measuring pesticidal activity are well known in the art. See, for example, Czapla and Lang (1990) J. Econ. Entomol. 83:2480-2485; Andrews et al. (1988) Biochem. J. 252:199-206; Marrone et al. (1985) J. of Economic Entomology 78:290-293; and U.S. Pat. No. 5,743,477, all of which are herein incorporated by reference in their entirety. As used here, a fragment comprises at least 8 contiguous amino acids of SEQ ID NO:2, 4, 6, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37 or 61. The invention encompasses other fragments, however, such as any fragment in the protein greater than about 10, 20, 30, 50, 100, 150, 200, 250, or 300 amino acids.
[0052] By "variants" is intended proteins or polypeptides having an amino acid sequence that is at least about 60%, 65%, about 70%, 75%, about 80%, 85%, about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO:2, 4, 6, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37 or 61. Variants also include polypeptides encoded by a nucleic acid molecule that hybridizes to the nucleic acid molecule of SEQ ID NO:1, 3, 5, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, or 60, or a complement thereof, under stringent conditions. Variants include polypeptides that differ in amino acid sequence due to mutagenesis. Variant proteins encompassed by the present invention are biologically active, that is they continue to possess the desired biological activity of the native protein, that is, retaining pesticidal activity. Methods for measuring pesticidal activity are well known in the art. See, for example, Czapla and Lang (1990) J. Econ. Entomol. 83:2480-2485; Andrews et al. (1988) Biochem. J. 252:199-206; Marrone et al. (1985) J. of Economic Entomology 78:290-293; and U.S. Pat. No. 5,743,477, all of which are herein incorporated by reference in their entirety.
[0053] Bacterial genes, such as the axmi genes of this invention, quite often possess multiple methionine initiation codons in proximity to the start of the open reading frame. Often, translation initiation at one or more of these start codons will lead to generation of a functional protein. These start codons can include ATG codons. However, bacteria such as Bacillus sp. also recognize the codon GTG as a start codon, and proteins that initiate translation at GTG codons contain a methionine at the first amino acid. Furthermore, it is not often determined a priori which of these codons are used naturally in the bacterium. Thus, it is understood that use of one of the alternate methionine codons may also lead to generation of pesticidal proteins. These pesticidal proteins are encompassed in the present invention and may be used in the methods of the present invention.
[0054] Antibodies to the polypeptides of the present invention, or to variants or fragments thereof, are also encompassed. Methods for producing antibodies are well known in the art (see, for example, Harlow and Lane (1988) Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.; U.S. Pat. No. 4,196,265).
Altered or Improved Variants
[0055] It is recognized that DNA sequences of a pesticidal protein may be altered by various methods, and that these alterations may result in DNA sequences encoding proteins with amino acid sequences different than that encoded by a pesticidal protein of the present invention. This protein may be altered in various ways including amino acid substitutions, deletions, truncations, and insertions of one or more amino acids of SEQ ID NO:2, 4, 6, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37 or 61, including up to about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 15, about 20, about 25, 30, a 30, about 35, about 40, 45, a 45, about 50, about 55, about 60, about 65, about 70, about 75, about 80, about 85, about 90, about 100, about 105, about 110, about 115, about 120, about 125, about 130, about 135, about 140, about 145, about 150, about 155, or more amino acid substitutions, deletions or insertions. Methods for such manipulations are generally known in the art. For example, amino acid sequence variants of a pesticidal protein can be prepared by mutations in the DNA. This may also be accomplished by one of several forms of mutagenesis and/or in directed evolution. In some aspects, the changes encoded in the amino acid sequence will not substantially affect the function of the protein. Such variants will possess the desired pesticidal activity. However, it is understood that the ability of a pesticidal protein to confer pesticidal activity may be improved by the use of such techniques upon the compositions of this invention. For example, one may express a pesticidal protein in host cells that exhibit high rates of base misincorporation during DNA replication, such as XL-1 Red (Stratagene, La Jolla, Calif.). After propagation in such strains, one can isolate the DNA (for example by preparing plasmid DNA, or by amplifying by PCR and cloning the resulting PCR fragment into a vector), culture the pesticidal protein mutations in a non-mutagenic strain, and identify mutated genes with pesticidal activity, for example by performing an assay to test for pesticidal activity. Generally, the protein is mixed and used in feeding assays. See, for example Marrone et al. (1985) J. of Economic Entomology 78:290-293. Such assays can include contacting plants with one or more pests and determining the plant's ability to survive and/or cause the death of the pests. Examples of mutations that result in increased toxicity are found in Schnepf et al. (1998) Microbiol. Mol. Biol. Rev. 62:775-806.
[0056] Alternatively, alterations may be made to the protein sequence of many proteins at the amino or carboxy terminus without substantially affecting activity. This can include insertions, deletions, or alterations introduced by modern molecular methods, such as PCR, including PCR amplifications that alter or extend the protein coding sequence by virtue of inclusion of amino acid encoding sequences in the oligonucleotides utilized in the PCR amplification. Alternatively, the protein sequences added can include entire protein-coding sequences, such as those used commonly in the art to generate protein fusions. Such fusion proteins are often used to (1) increase expression of a protein of interest (2) introduce a binding domain, enzymatic activity, or epitope to facilitate either protein purification, protein detection, or other experimental uses known in the art (3) target secretion or translation of a protein to a subcellular organelle, such as the periplasmic space of Gram-negative bacteria, or the endoplasmic reticulum of eukaryotic cells, the latter of which often results in glycosylation of the protein.
[0057] Variant nucleotide and amino acid sequences of the present invention also encompass sequences derived from mutagenic and recombinogenic procedures such as DNA shuffling. With such a procedure, one or more different pesticidal protein coding regions can be used to create a new pesticidal protein possessing the desired properties. In this manner, libraries of recombinant polynucleotides are generated from a population of related sequence polynucleotides comprising sequence regions that have substantial sequence identity and can be homologously recombined in vitro or in vivo. For example, using this approach, sequence motifs encoding a domain of interest may be shuffled between a pesticidal gene of the invention and other known pesticidal genes to obtain a new gene coding for a protein with an improved property of interest, such as an increased insecticidal activity. Strategies for such DNA shuffling are known in the art. See, for example, Stemmer (1994) Proc. Natl. Acad. Sci. USA 91:10747-10751; Stemmer (1994) Nature 370:389-391; Crameri et al. (1997) Nature Biotech. 15:436-438; Moore et al. (1997) J. Mol. Biol. 272:336-347; Zhang et al. (1997) Proc. Natl. Acad. Sci. USA 94:4504-4509; Crameri et al. (1998) Nature 391:288-291; and U.S. Pat. Nos. 5,605,793 and 5,837,458.
[0058] Domain swapping or shuffling is another mechanism for generating altered pesticidal proteins. Domains may be swapped between pesticidal proteins, resulting in hybrid or chimeric toxins with improved pesticidal activity or target spectrum. Methods for generating recombinant proteins and testing them for pesticidal activity are well known in the art (see, for example, Naimov et al. (2001) Appl. Environ. Microbiol. 67:5328-5330; de Maagd et al. (1996) Appl. Environ. Microbiol. 62:1537-1543; Ge et al. (1991) J. Biol. Chem. 266:17954-17958; Schnepf et al. (1990) J. Biol. Chem. 265:20923-20930; Rang et al. 91999) Appl. Environ. Microbiol. 65:2918-2925).
Vectors
[0059] A pesticidal sequence of the invention may be provided in an expression cassette for expression in a plant of interest. By "plant expression cassette" is intended a DNA construct that is capable of resulting in the expression of a protein from an open reading frame in a plant cell. Typically these contain a promoter and a coding sequence. Often, such constructs will also contain a 3' untranslated region. Such constructs may contain a "signal sequence" or "leader sequence" to facilitate co-translational or post-translational transport of the peptide to certain intracellular structures such as the chloroplast (or other plastid), endoplasmic reticulum, or Golgi apparatus.
[0060] By "signal sequence" is intended a sequence that is known or suspected to result in cotranslational or post-translational peptide transport across the cell membrane. In eukaryotes, this typically involves secretion into the Golgi apparatus, with some resulting glycosylation. Insecticidal toxins of bacteria are often synthesized as protoxins, which are protolytically activated in the gut of the target pest (Chang (1987) Methods Enzymol. 153:507-516). In some embodiments of the present invention, the signal sequence is located in the native sequence, or may be derived from a sequence of the invention. By "leader sequence" is intended any sequence that when translated, results in an amino acid sequence sufficient to trigger co-translational transport of the peptide chain to a subcellular organelle. Thus, this includes leader sequences targeting transport and/or glycosylation by passage into the endoplasmic reticulum, passage to vacuoles, plastids including chloroplasts, mitochondria, and the like.
[0061] By "plant transformation vector" is intended a DNA molecule that is necessary for efficient transformation of a plant cell. Such a molecule may consist of one or more plant expression cassettes, and may be organized into more than one "vector" DNA molecule. For example, binary vectors are plant transformation vectors that utilize two non-contiguous DNA vectors to encode all requisite cis- and trans-acting functions for transformation of plant cells (Hellens and Mullineaux (2000) Trends in Plant Science 5:446-451). "Vector" refers to a nucleic acid construct designed for transfer between different host cells. "Expression vector" refers to a vector that has the ability to incorporate, integrate and express heterologous DNA sequences or fragments in a foreign cell. The cassette will include 5' and 3' regulatory sequences operably linked to a sequence of the invention. By "operably linked" is intended a functional linkage between a promoter and a second sequence, wherein the promoter sequence initiates and mediates transcription of the DNA sequence corresponding to the second sequence. Generally, operably linked means that the nucleic acid sequences being linked are contiguous and, where necessary to join two protein coding regions, contiguous and in the same reading frame. The cassette may additionally contain at least one additional gene to be cotransformed into the organism. Alternatively, the additional gene(s) can be provided on multiple expression cassettes.
[0062] "Promoter" refers to a nucleic acid sequence that functions to direct transcription of a downstream coding sequence. The promoter together with other transcriptional and translational regulatory nucleic acid sequences (also termed "control sequences") are necessary for the expression of a DNA sequence of interest.
[0063] Such an expression cassette is provided with a plurality of restriction sites for insertion of the pesticidal sequence to be under the transcriptional regulation of the regulatory regions.
[0064] The expression cassette will include in the 5'-3' direction of transcription, a transcriptional and translational initiation region (i.e., a promoter), a DNA sequence of the invention, and a translational and transcriptional termination region (i.e., termination region) functional in plants. The promoter may be native or analogous, or foreign or heterologous, to the plant host and/or to the DNA sequence of the invention. Additionally, the promoter may be the natural sequence or alternatively a synthetic sequence. Where the promoter is "native" or "homologous" to the plant host, it is intended that the promoter is found in the native plant into which the promoter is introduced. Where the promoter is "foreign" or "heterologous" to the DNA sequence of the invention, it is intended that the promoter is not the native or naturally occurring promoter for the operably linked DNA sequence of the invention.
[0065] The termination region may be native with the transcriptional initiation region, may be native with the operably linked DNA sequence of interest, may be native with the plant host, or may be derived from another source (i.e., foreign or heterologous to the promoter, the DNA sequence of interest, the plant host, or any combination thereof). Convenient termination regions are available from the Ti-plasmid of A. tumefaciens, such as the octopine synthase and nopaline synthase termination regions. See also Guerineau et al. (1991) Mol. Gen. Genet. 262:141-144; Proudfoot (1991) Cell 64:671-674; Sanfacon et al. (1991) Genes Dev. 5:141-149; Mogen et al. (1990) Plant Cell 2:1261-1272; Munroe et al. (1990) Gene 91:151-158; Ballas et al. (1989) Nucleic Acids Res. 17:7891-7903; and Joshi et al. (1987) Nucleic Acid Res. 15:9627-9639.
[0066] Where appropriate, the gene(s) may be optimized for increased expression in the transformed host cell. That is, the genes can be synthesized using host cell-preferred codons for improved expression, or may be synthesized using codons at a host-preferred codon usage frequency. Generally, the GC content of the gene will be increased. See, for example, Campbell and Gowri (1990) Plant Physiol. 92:1-11 for a discussion of host-preferred codon usage. Methods are available in the art for synthesizing plant-preferred genes. See, for example, U.S. Pat. Nos. 5,380,831, and 5,436,391, and Murray et al. (1989) Nucleic Acids Res. 17:477-498, herein incorporated by reference.
[0067] In one embodiment, the pesticidal protein is targeted to the chloroplast for expression. In this manner, where the pesticidal protein is not directly inserted into the chloroplast, the expression cassette will additionally contain a nucleic acid encoding a transit peptide to direct the pesticidal protein to the chloroplasts. Such transit peptides are known in the art. See, for example, Von Heijne et al. (1991) Plant Mol. Biol. Rep. 9:104-126; Clark et al. (1989) J. Biol. Chem. 264:17544-17550; Della-Cioppa et al. (1987) Plant Physiol. 84:965-968; Romer et al. (1993) Biochem. Biophys. Res. Commun. 196:1414-1421; and Shah et al. (1986) Science 233:478-481.
[0068] The pesticidal gene to be targeted to the chloroplast may be optimized for expression in the chloroplast to account for differences in codon usage between the plant nucleus and this organelle. In this manner, the nucleic acids of interest may be synthesized using chloroplast-preferred codons. See, for example, U.S. Pat. No. 5,380,831, herein incorporated by reference.
Plant Transformation
[0069] Methods of the invention involve introducing a nucleotide construct into a plant. By "introducing" is intended to present to the plant the nucleotide construct in such a manner that the construct gains access to the interior of a cell of the plant. The methods of the invention do not require that a particular method for introducing a nucleotide construct to a plant is used, only that the nucleotide construct gains access to the interior of at least one cell of the plant. Methods for introducing nucleotide constructs into plants are known in the art including, but not limited to, stable transformation methods, transient transformation methods, and virus-mediated methods.
[0070] By "plant" is intended whole plants, plant organs (e.g., leaves, stems, roots, etc.), seeds, plant cells, propagules, embryos and progeny of the same. Plant cells can be differentiated or undifferentiated (e.g. callus, suspension culture cells, protoplasts, leaf cells, root cells, phloem cells, pollen).
[0071] "Transgenic plants" or "transformed plants" or "stably transformed" plants or cells or tissues refers to plants that have incorporated or integrated exogenous nucleic acid sequences or DNA fragments into the plant cell. These nucleic acid sequences include those that are exogenous, or not present in the untransformed plant cell, as well as those that may be endogenous, or present in the untransformed plant cell.
[0072] "Heterologous" generally refers to the nucleic acid sequences that are not endogenous to the cell or part of the native genome in which they are present, and have been added to the cell by infection, transfection, microinjection, electroporation, microprojection, or the like.
[0073] Transformation of plant cells can be accomplished by one of several techniques known in the art. The pesticidal gene of the invention may be modified to obtain or enhance expression in plant cells. Typically a construct that expresses such a protein would contain a promoter to drive transcription of the gene, as well as a 3' untranslated region to allow transcription termination and polyadenylation. The organization of such constructs is well known in the art. In some instances, it may be useful to engineer the gene such that the resulting peptide is secreted, or otherwise targeted within the plant cell. For example, the gene can be engineered to contain a signal peptide to facilitate transfer of the peptide to the endoplasmic reticulum. It may also be preferable to engineer the plant expression cassette to contain an intron, such that mRNA processing of the intron is required for expression.
[0074] Typically this "plant expression cassette" will be inserted into a "plant transformation vector". This plant transformation vector may be comprised of one or more DNA vectors needed for achieving plant transformation. For example, it is a common practice in the art to utilize plant transformation vectors that are comprised of more than one contiguous DNA segment. These vectors are often referred to in the art as "binary vectors". Binary vectors as well as vectors with helper plasmids are most often used for Agrobacterium-mediated transformation, where the size and complexity of DNA segments needed to achieve efficient transformation is quite large, and it is advantageous to separate functions onto separate DNA molecules. Binary vectors typically contain a plasmid vector that contains the cis-acting sequences required for T-DNA transfer (such as left border and right border), a selectable marker that is engineered to be capable of expression in a plant cell, and a "gene of interest" (a gene engineered to be capable of expression in a plant cell for which generation of transgenic plants is desired). Also present on this plasmid vector are sequences required for bacterial replication. The cis-acting sequences are arranged in a fashion to allow efficient transfer into plant cells and expression therein. For example, the selectable marker gene and the pesticidal gene are located between the left and right borders. Often a second plasmid vector contains the trans-acting factors that mediate T-DNA transfer from Agrobacterium to plant cells. This plasmid often contains the virulence functions (Vir genes) that allow infection of plant cells by Agrobacterium, and transfer of DNA by cleavage at border sequences and vir-mediated DNA transfer, as is understood in the art (Hellens and Mullineaux (2000) Trends in Plant Science 5:446-451). Several types of Agrobacterium strains (e.g. LBA4404, GV3101, EHA101, EHA105, etc.) can be used for plant transformation. The second plasmid vector is not necessary for transforming the plants by other methods such as microprojection, microinjection, electroporation, polyethylene glycol, etc.
[0075] In general, plant transformation methods involve transferring heterologous DNA into target plant cells (e.g. immature or mature embryos, suspension cultures, undifferentiated callus, protoplasts, etc.), followed by applying a maximum threshold level of appropriate selection (depending on the selectable marker gene) to recover the transformed plant cells from a group of untransformed cell mass. Explants are typically transferred to a fresh supply of the same medium and cultured routinely. Subsequently, the transformed cells are differentiated into shoots after placing on regeneration medium supplemented with a maximum threshold level of selecting agent. The shoots are then transferred to a selective rooting medium for recovering rooted shoot or plantlet. The transgenic plantlet then grows into a mature plant and produces fertile seeds (e.g. Hiei et al. (1994) The Plant Journal 6:271-282; Ishida et al. (1996) Nature Biotechnology 14:745-750). Explants are typically transferred to a fresh supply of the same medium and cultured routinely. A general description of the techniques and methods for generating transgenic plants are found in Ayres and Park (1994) Critical Reviews in Plant Science 13:219-239 and Bommineni and Jauhar (1997) Maydica 42:107-120. Since the transformed material contains many cells; both transformed and non-transformed cells are present in any piece of subjected target callus or tissue or group of cells. The ability to kill non-transformed cells and allow transformed cells to proliferate results in transformed plant cultures. Often, the ability to remove non-transformed cells is a limitation to rapid recovery of transformed plant cells and successful generation of transgenic plants.
[0076] Transformation protocols as well as protocols for introducing nucleotide sequences into plants may vary depending on the type of plant or plant cell, i.e., monocot or dicot, targeted for transformation. Generation of transgenic plants may be performed by one of several methods, including, but not limited to, microinjection, electroporation, direct gene transfer, introduction of heterologous DNA by Agrobacterium into plant cells (Agrobacterium-mediated transformation), bombardment of plant cells with heterologous foreign DNA adhered to particles, ballistic particle acceleration, aerosol beam transformation (U.S. Published Application No. 20010026941; U.S. Pat. No. 4,945,050; International Publication No. WO 91/00915; U.S. Published Application No. 2002015066), Lec1 transformation, and various other non-particle direct-mediated methods to transfer DNA.
[0077] Methods for transformation of chloroplasts are known in the art. See, for example, Svab et al. (1990) Proc. Natl. Acad. Sci. USA 87:8526-8530; Svab and Maliga (1993) Proc. Natl. Acad. Sci. USA 90:913-917; Svab and Maliga (1993) EMBO J. 12:601-606. The method relies on particle gun delivery of DNA containing a selectable marker and targeting of the DNA to the plastid genome through homologous recombination. Additionally, plastid transformation can be accomplished by transactivation of a silent plastid-borne transgene by tissue-preferred expression of a nuclear-encoded and plastid-directed RNA polymerase. Such a system has been reported in McBride et al. (1994) Proc. Natl. Acad. Sci. USA 91:7301-7305.
[0078] Following integration of heterologous foreign DNA into plant cells, one then applies a maximum threshold level of appropriate selection in the medium to kill the untransformed cells and separate and proliferate the putatively transformed cells that survive from this selection treatment by transferring regularly to a fresh medium. By continuous passage and challenge with appropriate selection, one identifies and proliferates the cells that are transformed with the plasmid vector. Molecular and biochemical methods can then be used to confirm the presence of the integrated heterologous gene of interest into the genome of the transgenic plant.
[0079] The cells that have been transformed may be grown into plants in accordance with conventional ways. See, for example, McCormick et al. (1986) Plant Cell Reports 5:81-84. These plants may then be grown, and either pollinated with the same transformed strain or different strains, and the resulting hybrid having constitutive expression of the desired phenotypic characteristic identified. Two or more generations may be grown to ensure that expression of the desired phenotypic characteristic is stably maintained and inherited and then seeds harvested to ensure expression of the desired phenotypic characteristic has been achieved. In this manner, the present invention provides transformed seed (also referred to as "transgenic seed") having a nucleotide construct of the invention, for example, an expression cassette of the invention, stably incorporated into their genome.
Evaluation of Plant Transformation
[0080] Following introduction of heterologous foreign DNA into plant cells, the transformation or integration of heterologous gene in the plant genome is confirmed by various methods such as analysis of nucleic acids, proteins and metabolites associated with the integrated gene.
[0081] PCR analysis is a rapid method to screen transformed cells, tissue or shoots for the presence of incorporated gene at the earlier stage before transplanting into the soil (Sambrook and Russell (2001) Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.). PCR is carried out using oligonucleotide primers specific to the gene of interest or Agrobacterium vector background, etc.
[0082] Plant transformation may be confirmed by Southern blot analysis of genomic DNA (Sambrook and Russell, 2001, supra). In general, total DNA is extracted from the transformant, digested with appropriate restriction enzymes, fractionated in an agarose gel and transferred to a nitrocellulose or nylon membrane. The membrane or "blot" is then probed with, for example, radiolabeled 32P target DNA fragment to confirm the integration of introduced gene into the plant genome according to standard techniques (Sambrook and Russell, 2001, supra).
[0083] In Northern blot analysis, RNA is isolated from specific tissues of transformant, fractionated in a formaldehyde agarose gel, and blotted onto a nylon filter according to standard procedures that are routinely used in the art (Sambrook and Russell, 2001, supra). Expression of RNA encoded by the pesticidal gene is then tested by hybridizing the filter to a radioactive probe derived from a pesticidal gene, by methods known in the art (Sambrook and Russell, 2001, supra).
[0084] Western blot, biochemical assays and the like may be carried out on the transgenic plants to confirm the presence of protein encoded by the pesticidal gene by standard procedures (Sambrook and Russell, 2001, supra) using antibodies that bind to one or more epitopes present on the pesticidal protein.
Pesticidal Activity in Plants
[0085] In another aspect of the invention, one may generate transgenic plants expressing a pesticidal protein that has pesticidal activity. Methods described above by way of example may be utilized to generate transgenic plants, but the manner in which the transgenic plant cells are generated is not critical to this invention. Methods known or described in the art such as Agrobacterium-mediated transformation, biolistic transformation, and non-particle-mediated methods may be used at the discretion of the experimenter. Plants expressing a pesticidal protein may be isolated by common methods described in the art, for example by transformation of callus, selection of transformed callus, and regeneration of fertile plants from such transgenic callus. In such process, one may use any gene as a selectable marker so long as its expression in plant cells confers ability to identify or select for transformed cells.
[0086] A number of markers have been developed for use with plant cells, such as resistance to chloramphenicol, the aminoglycoside G418, hygromycin, or the like. Other genes that encode a product involved in chloroplast metabolism may also be used as selectable markers. For example, genes that provide resistance to plant herbicides such as glyphosate, bromoxynil, or imidazolinone may find particular use. Such genes have been reported (Stalker et al. (1985) J. Biol. Chem. 263:6310-6314 (bromoxynil resistance nitrilase gene); and Sathasivan et al. (1990) Nucl. Acids Res. 18:2188 (AHAS imidazolinone resistance gene). Additionally, the genes disclosed herein are useful as markers to assess transformation of bacterial or plant cells. Methods for detecting the presence of a transgene in a plant, plant organ (e.g., leaves, stems, roots, etc.), seed, plant cell, propagule, embryo or progeny of the same are well known in the art. In one embodiment, the presence of the transgene is detected by testing for pesticidal activity.
[0087] Fertile plants expressing a pesticidal protein may be tested for pesticidal activity, and the plants showing optimal activity selected for further breeding. Methods are available in the art to assay for pest activity. Generally, the protein is mixed and used in feeding assays. See, for example Marrone et al. (1985) J. of Economic Entomology 78:290-293.
[0088] The present invention may be used for transformation of any plant species, including, but not limited to, monocots and dicots. Examples of plants of interest include, but are not limited to, corn (maize), sorghum, wheat, sunflower, tomato, crucifers, peppers, potato, cotton, rice, soybean, sugarbeet, sugarcane, tobacco, barley, and oilseed rape, Brassica sp., alfalfa, rye, millet, safflower, peanuts, sweet potato, cassava, coffee, coconut, pineapple, citrus trees, cocoa, tea, banana, avocado, fig, guava, mango, olive, papaya, cashew, macadamia, almond, oats, vegetables, ornamentals, and conifers.
[0089] Vegetables include, but are not limited to, tomatoes, lettuce, green beans, lima beans, peas, and members of the genus Curcumis such as cucumber, cantaloupe, and musk melon. Ornamentals include, but are not limited to, azalea, hydrangea, hibiscus, roses, tulips, daffodils, petunias, carnation, poinsettia, and chrysanthemum. Preferably, plants of the present invention are crop plants (for example, maize, sorghum, wheat, sunflower, tomato, crucifers, peppers, potato, cotton, rice, soybean, sugarbeet, sugarcane, tobacco, barley, oilseed rape., etc.).
Use in Pesticidal Control
[0090] General methods for employing strains comprising a nucleotide sequence of the present invention, or a variant thereof, in pesticide control or in engineering other organisms as pesticidal agents are known in the art. See, for example U.S. Pat. No. 5,039,523 and EP 0480762A2.
[0091] The Bacillus strains containing a nucleotide sequence of the present invention, or a variant thereof, or the microorganisms that have been genetically altered to contain a pesticidal gene and protein may be used for protecting agricultural crops and products from pests. In one aspect of the invention, whole, i.e., unlysed, cells of a toxin (pesticide)-producing organism are treated with reagents that prolong the activity of the toxin produced in the cell when the cell is applied to the environment of target pest(s).
[0092] Alternatively, the pesticide is produced by introducing a pesticidal gene into a cellular host. Expression of the pesticidal gene results, directly or indirectly, in the intracellular production and maintenance of the pesticide. In one aspect of this invention, these cells are then treated under conditions that prolong the activity of the toxin produced in the cell when the cell is applied to the environment of target pest(s). The resulting product retains the toxicity of the toxin. These naturally encapsulated pesticides may then be formulated in accordance with conventional techniques for application to the environment hosting a target pest, e.g., soil, water, and foliage of plants. See, for example EPA 0192319, and the references cited therein. Alternatively, one may formulate the cells expressing a gene of this invention such as to allow application of the resulting material as a pesticide.
[0093] The active ingredients of the present invention are normally applied in the form of compositions and can be applied to the crop area or plant to be treated, simultaneously or in succession, with other compounds. These compounds can be fertilizers, weed killers, cryoprotectants, surfactants, detergents, pesticidal soaps, dormant oils, polymers, and/or time-release or biodegradable carrier formulations that permit long-term dosing of a target area following a single application of the formulation. They can also be selective herbicides, chemical insecticides, virucides, microbicides, amoebicides, pesticides, fungicides, bacteriocides, nematocides, molluscicides or mixtures of several of these preparations, if desired, together with further agriculturally acceptable carriers, surfactants or application-promoting adjuvants customarily employed in the art of formulation. Suitable carriers and adjuvants can be solid or liquid and correspond to the substances ordinarily employed in formulation technology, e.g. natural or regenerated mineral substances, solvents, dispersants, wetting agents, tackifiers, binders or fertilizers. Likewise the formulations may be prepared into edible "baits" or fashioned into pest "traps" to permit feeding or ingestion by a target pest of the pesticidal formulation.
[0094] Methods of applying an active ingredient of the present invention or an agrochemical composition of the present invention that contains at least one of the pesticidal proteins produced by the bacterial strains of the present invention include leaf application, seed coating and soil application. The number of applications and the rate of application depend on the intensity of infestation by the corresponding pest.
[0095] The composition may be formulated as a powder, dust, pellet, granule, spray, emulsion, colloid, solution, or such like, and may be prepared by such conventional means as desiccation, lyophilization, homogenation, extraction, filtration, centrifugation, sedimentation, or concentration of a culture of cells comprising the polypeptide. In all such compositions that contain at least one such pesticidal polypeptide, the polypeptide may be present in a concentration of from about 1% to about 99% by weight.
[0096] Lepidopteran, dipteran, or coleopteran pests may be killed or reduced in numbers in a given area by the methods of the invention, or may be prophylactically applied to an environmental area to prevent infestation by a susceptible pest. Preferably the pest ingests, or is contacted with, a pesticidally-effective amount of the polypeptide. By "pesticidally-effective amount" is intended an amount of the pesticide that is able to bring about death to at least one pest, or to noticeably reduce pest growth, feeding, or normal physiological development. This amount will vary depending on such factors as, for example, the specific target pests to be controlled, the specific environment, location, plant, crop, or agricultural site to be treated, the environmental conditions, and the method, rate, concentration, stability, and quantity of application of the pesticidally-effective polypeptide composition. The formulations may also vary with respect to climatic conditions, environmental considerations, and/or frequency of application and/or severity of pest infestation.
[0097] The pesticide compositions described may be made by formulating either the bacterial cell, crystal and/or spore suspension, or isolated protein component with the desired agriculturally-acceptable carrier. The compositions may be formulated prior to administration in an appropriate means such as lyophilized, freeze-dried, desiccated, or in an aqueous carrier, medium or suitable diluent, such as saline or other buffer. The formulated compositions may be in the form of a dust or granular material, or a suspension in oil (vegetable or mineral), or water or oil/water emulsions, or as a wettable powder, or in combination with any other carrier material suitable for agricultural application. Suitable agricultural carriers can be solid or liquid and are well known in the art. The term "agriculturally-acceptable carrier" covers all adjuvants, inert components, dispersants, surfactants, tackifiers, binders, etc. that are ordinarily used in pesticide formulation technology; these are well known to those skilled in pesticide formulation. The formulations may be mixed with one or more solid or liquid adjuvants and prepared by various means, e.g., by homogeneously mixing, blending and/or grinding the pesticidal composition with suitable adjuvants using conventional formulation techniques. Suitable formulations and application methods are described in U.S. Pat. No. 6,468,523, herein incorporated by reference.
[0098] "Pest" includes but is not limited to, insects, fungi, bacteria, nematodes, mites, ticks, and the like. Insect pests include insects selected from the orders Coleoptera, Diptera, Hymenoptera, Lepidoptera, Mallophaga, Homoptera, Hemiptera, Orthroptera, Thysanoptera, Dermaptera, Isoptera, Anoplura, Siphonaptera, Trichoptera, etc., particularly Coleoptera, Lepidoptera, and Diptera.
[0099] The order Coleoptera includes the suborders Adephaga and Polyphaga. Suborder Adephaga includes the superfamilies Caraboidea and Gyrinoidea, while suborder Polyphaga includes the superfamilies Hydrophiloidea, Staphylinoidea, Cantharoidea, Cleroidea, Elateroidea, Dascilloidea, Dryopoidea, Byrrhoidea, Cucujoidea, Meloidea, Mordelloidea, Tenebrionoidea, Bostrichoidea, Scarabaeoidea, Cerambycoidea, Chrysomeloidea, and Curculionoidea. Superfamily Caraboidea includes the families Cicindelidae, Carabidae, and Dytiscidae. Superfamily Gyrinoidea includes the family Gyrimidae. Superfamily Hydrophiloidea includes the family Hydrophilidae. Superfamily Staphylinoidea includes the families Silphidae and Staphylimidae. Superfamily Cantharoidea includes the families Cantharidae and Lampyridae. Superfamily Cleroidea includes the families Cleridae and Dermestidae. Superfamily Elateroidea includes the families Elateridae and Buprestidae. Superfamily Cucujoidea includes the family Coccinellidae. Superfamily Meloidea includes the family Meloidae. Superfamily Tenebrionoidea includes the family Tenebrionidae. Superfamily Scarabaeoidea includes the families Passalidae and Scarabaeidae. Superfamily Cerambycoidea includes the family Cerambycidae. Superfamily Chrysomeloidea includes the family Chrysomelidae. Superfamily Curculionoidea includes the families Curculionidae and Scolytidae.
[0100] The order Diptera includes the Suborders Nematocera, Brachycera, and Cyclorrhapha. Suborder Nematocera includes the families Tipulidae, Psychodidae, Culicidae, Ceratopogonidae, Chironomidae, Simuliidae, Bibionidae, and Cecidomyiidae. Suborder Brachycera includes the families Stratiomyidae, Tabanidae, Therevidae, Asilidae, Mydidae, Bombyliidae, and Dolichopodidae. Suborder Cyclorrhapha includes the Divisions Aschiza and Aschiza. Division Aschiza includes the families Phoridae, Syrphidae, and Conopidae. Division Aschiza includes the Sections Acalyptratae and Calyptratae. Section Acalyptratae includes the families Otitidae, Tephritidae, Agromyzidae, and Drosophilidae. Section Calyptratae includes the families Hippoboscidae, Oestridae, Tachimidae, Anthomyiidae, Muscidae, Calliphoridae, and Sarcophagidae.
[0101] The order Lepidoptera includes the families Papilionidae, Pieridae, Lycaenidae, Nymphalidae, Danaidae, Satyridae, Hesperiidae, Sphingidae, Saturniidae, Geometridae, Arctiidae, Noctuidae, Lymantriidae, Sesiidae, and Tineidae.
[0102] Insect pests of the invention for the major crops include: Maize: Ostrinia nubilalis, European corn borer; Agrotis ipsilon, black cutworm; Helicoverpa zea, corn earworm; Spodoptera frugiperda, fall armyworm; Diatraea grandiosella, southwestern corn borer; Elasmopalpus lignosellus, lesser cornstalk borer; Diatraea saccharalis, surgarcane borer; Diabrotica virgifera, western corn rootworm; Diabrotica longicornis barberi, northern corn rootworm; Diabrotica undecimpunctata howardi, southern corn rootworm; Melanotus spp., wireworms; Cyclocephala borealis, northern masked chafer (white grub); Cyclocephala immaculata, southern masked chafer (white grub); Popillia japonica, Japanese beetle; Chaetocnema pulicaria, corn flea beetle; Sphenophorus maidis, maize billbug; Rhopalosiphum maidis, corn leaf aphid; Anuraphis maidiradicis, corn root aphid; Blissus leucopterus leucopterus, chinch bug; Melanoplus femurrubrum, redlegged grasshopper; Melanoplus sanguinipes, migratory grasshopper; Hylemya platura, seedcorn maggot; Agromyza parvicornis, corn blot leafminer; Anaphothrips obscrurus, grass thrips; Solenopsis milesta, thief ant; Tetranychus urticae, twospotted spider mite; Sorghum: Chilo partellus, sorghum borer; Spodoptera frugiperda, fall armyworm; Helicoverpa zea, corn earworm; Elasmopalpus lignosellus, lesser cornstalk borer; Feltia subterranea, granulate cutworm; Phyllophaga crinita, white grub; Eleodes, Conoderus, and Aeolus spp., wireworms; Oulema melanopus, cereal leaf beetle; Chaetocnema pulicaria, corn flea beetle; Sphenophorus maidis, maize billbug; Rhopalosiphum maidis; corn leaf aphid; Sipha flava, yellow sugarcane aphid; Blissus leucopterus leucopterus, chinch bug; Contarinia sorghicola, sorghum midge; Tetranychus cinnabarinus, carmine spider mite; Tetranychus urticae, twospotted spider mite Wheat: Pseudaletia unipunctata, army worm; Spodoptera frugiperda, fall armyworm; Elasmopalpus lignosellus, lesser cornstalk borer; Agrotis orthogonia, western cutworm; Elasmopalpus lignosellus, lesser cornstalk borer; Oulema melanopus, cereal leaf beetle; Hypera punctata, clover leaf weevil; Diabrotica undecimpunctata howardi, southern corn rootworm; Russian wheat aphid; Schizaphis graminum, greenbug; Macrosiphum avenae, English grain aphid; Melanoplus femurrubrum, redlegged grasshopper; Melanoplus differentialis, differential grasshopper; Melanoplus sanguinipes, migratory grasshopper; Mayetiola destructor, Hessian fly; Sitodiplosis mosellana, wheat midge; Meromyza americana, wheat stem maggot; Hylemya coarctata, wheat bulb fly; Frankliniella fusca, tobacco thrips; Cephus cinctus, wheat stem sawfly; Aceria tulipae, wheat curl mite; Sunflower: Suleima helianthana, sunflower bud moth; Homoeosoma electellum, sunflower moth; zygogramma exclamationis, sunflower beetle; Bothyrus gibbosus, carrot beetle; Neolasioptera murtfeldtiana, sunflower seed midge; Cotton: Heliothis virescens, cotton budworm; Helicoverpa zea, cotton bollworm; Spodoptera exigua, beet armyworm; Pectinophora gossypiella, pink bollworm; Anthonomus grandis, boll weevil; Aphis gossypii, cotton aphid; Pseudatomoscelis seriatus, cotton fleahopper; Trialeurodes abutilonea, bandedwinged whitefly; Lygus lineolaris, tarnished plant bug; Melanoplus femurrubrum, redlegged grasshopper; Melanoplus differentialis, differential grasshopper; Thrips tabaci, onion thrips; Franklinkiella fusca, tobacco thrips; Tetranychus cinnabarinus, carmine spider mite; Tetranychus urticae, twospotted spider mite; Rice: Diatraea saccharalis, sugarcane borer; Spodoptera frugiperda, fall armyworm; Helicoverpa zea, corn earworm; Colaspis brunnea, grape colaspis; Lissorhoptrus oryzophilus, rice water weevil; Sitophilus oryzae, rice weevil; Nephotettix nigropictus, rice leafhopper; Blissus leucopterus leucopterus, chinch bug; Acrosternum hilare, green stink bug; Soybean: Pseudoplusia includens, soybean looper; Anticarsia gemmatalis, velvetbean caterpillar; Plathypena scabra, green cloverworm; Ostrinia nubilalis, European corn borer; Agrotis ipsilon, black cutworm; Spodoptera exigua, beet armyworm; Heliothis virescens, cotton budworm; Helicoverpa zea, cotton bollworm; Epilachna varivestis, Mexican bean beetle; Myzus persicae, green peach aphid; Empoasca fabae, potato leafhopper; Acrosternum hilare, green stink bug; Melanoplus femurrubrum, redlegged grasshopper; Melanoplus differentialis, differential grasshopper; Hylemya platura, seedcorn maggot; Sericothrips variabilis, soybean thrips; Thrips tabaci, onion thrips; Tetranychus turkestani, strawberry spider mite; Tetranychus urticae, twospotted spider mite; Barley: Ostrinia nubilalis, European corn borer; Agrotis ipsilon, black cutworm; Schizaphis graminum, greenbug; Blissus leucopterus leucopterus, chinch bug; Acrosternum hilare, green stink bug; Euschistus servus, brown stink bug; Delia platura, seedcorn maggot; Mayetiola destructor, Hessian fly; Petrobia latens, brown wheat mite; Oil Seed Rape: Brevicoryne brassicae, cabbage aphid; Phyllotreta cruciferae, Flea beetle; Mamestra configurata, Bertha armyworm; Plutella xylostella, Diamond-back moth; Delia ssp., Root maggots.
[0103] Nematodes include parasitic nematodes such as root-knot, cyst, and lesion nematodes, including Heterodera spp., Meloidogyne spp., and Globodera spp.; particularly members of the cyst nematodes, including, but not limited to, Heterodera glycines (soybean cyst nematode); Heterodera schachtii (beet cyst nematode); Heterodera avenae (cereal cyst nematode); and Globodera rostochiensis and Globodera pailida (potato cyst nematodes). Lesion nematodes include Pratylenchus spp.
Methods for Increasing Plant Yield
[0104] Methods for increasing plant yield are provided. The methods comprise introducing into a plant or plant cell a polynucleotide comprising a pesticidal sequence disclosed herein. As defined herein, the "yield" of the plant refers to the quality and/or quantity of biomass produced by the plant. By "biomass" is intended any measured plant product. An increase in biomass production is any improvement in the yield of the measured plant product. Increasing plant yield has several commercial applications. For example, increasing plant leaf biomass may increase the yield of leafy vegetables for human or animal consumption. Additionally, increasing leaf biomass can be used to increase production of plant-derived pharmaceutical or industrial products. An increase in yield can comprise any statistically significant increase including, but not limited to, at least a 1% increase, at least a 3% increase, at least a 5% increase, at least a 10% increase, at least a 20% increase, at least a 30%, at least a 50%, at least a 70%, at least a 100% or a greater increase in yield compared to a plant not expressing the pesticidal sequence.
[0105] The following examples are offered by way of illustration and not by way of limitation.
EXPERIMENTAL
Example 1
Extraction of Plasmid DNA
[0106] Strains ATX14759, ATX14875, ATX13008, ATX13002, ATX9387, ATX13045, ATX21738, ATX14833, ATX1489, ATX15398 and ATX12972 were selected for analysis. Pure cultures of each strain were grown in large quantities of rich media. The cultures were centrifuged to harvest the cell pellet. The cell pellet was then prepared by treatment with SDS by methods known in the art, resulting in breakage of the cell wall and release of DNA. Proteins and large genomic DNA were then precipitated by a high salt concentration. The plasmid DNA was then precipitated with ethanol. In several instances, the plasmid DNA was separated from any remaining chromosomal DNA by high-speed centrifugation through a cesium chloride gradient. Alternatively, the plasmid DNA was purified by binding to a resin, as known in the art. For each strain, the quality of the DNA was checked by visualization on an agarose gel by methods known in the art.
Example 2
Cloning of Genes
[0107] DNA libraries were prepared from the plasmid DNA or each strain. This may be achieved in many ways as known in the art. For, example, the purified plasmid DNA can be sheared into 5-10 kb sized fragments and the 5' and 3' single stranded overhangs repaired using T4 DNA polymerase and Klenow fragment in the presence of all four dNTPs, as known in the art. Phosphates can then be attached to the 5' ends by treatment with T4 polynucleotide kinase, as known in the art. The repaired DNA fragments can then be ligated overnight into a standard high copy vector (i.e. pBLUESCRIPT® SK+), suitably prepared to accept the inserts as known in the art (for example by digestion with a restriction enzyme producing blunt ends).
[0108] The quality of the resulting DNA libraries was analyzed by digesting a subset of clones with a restriction enzyme known to have a cleavage site flanking the cloning site.
[0109] A high percentage of clones were determined to contain inserts, ideally with an average insert size of 5-6 kb.
Example 3
High Throughput Sequencing of Library Plates
[0110] Once the DNA library quality was checked and confirmed, colonies were grown in a rich broth in 2 ml 96-well blocks overnight at 37° C., typically at a shaking speed of 350 rpm. The blocks were centrifuged to harvest the cells to the bottom of the block. The blocks were then prepared by standard alkaline lysis prep in a high throughput format.
[0111] The end sequences of clones from this library were then determined for a large number of clones from each block in the following manner: The DNA sequence of each clone chosen for analysis was determined using the fluorescent dye terminator sequencing technique (Applied Biosystems), by methods known in the art using an automated DNA sequencing machine, and standard oligonucleotide primers that anneal to the plasmid vector in the region flanking the insert.
Example 4
Assembly and Screening of Sequencing Data
[0112] DNA sequences obtained were compiled into an assembly project and aligned together to form contigs. This can be done efficiently using a computer program, such as Vector NTI, or alternatively by using the Phred/Phrap suite of DNA alignment and analysis programs. These contigs, along with any individual read that may not have been added to a contig, were compared to a compiled database of all classes of known pesticidal genes. Contigs or individual reads identified as having identity to a known endotoxin or pesticidal gene were analyzed further.
Example 5
Axmi-037
[0113] From strain ATX1489, clone pAX2558 was found to contain an open reading frame with homology to "cry" type delta-endotoxins. This open reading frame was designated as axmi-037 (SEQ ID NO: 16). Inspection of the axmi-037 open reading frame suggests that more than one start codon may be present. The two predicted start codons are the ATG codon beginning at nucleotide position 1 of SEQ ID NO: 16, and a downstream ATG codon (represented by SEQ ID NO: 18). The ATG at nucleotide 77 of SEQ ID NO:16 has a ribosome binding site sequence (5'-G-G-A-G-G-3'), located at nucleotide positions 63-67 of SEQ ID NO: 16. Based on the presence of this strong ribosome binding consensus sequence immediately upstream of this second start site, and the homology of the two predicted proteins to other endotoxins, the translation product of the downstream start site is herein designated AXMI-037 (SEQ ID NO: 19). The longer translation product, beginning at the ATG at nucleotide position 1 of SEQ ID NO: 16, is designated AXMI-37-2 (and set forth in SEQ ID NO:17). pAX2558 was deposited with the ARS Patent Strain Collection on Jun. 15, 2006, and assigned NRRL B-30939. AXMI-37-2 exhibits 60% amino acid identity to the Cry7Aa1 endotoxin.
Example 6
Axmi-019
[0114] From strain ATX14875, a clone was found to contain an open reading frame with homology to MTX family toxins. This open reading frame was designated as axmi-019 (SEQ ID NO:10), and the encoded protein was designated AXMI-019 (SEQ ID NO:11). By searching of public databases of protein sequences, such as the GenPept database, the C-terminal region of AXMI-019 (starting at approximately amino acid 123 of SEQ ID NO: 11) was found to have low homology to a class of toxins including Bacillus thuringiensis serovar darmstadiensis Cry14-4 toxin (SEQ ID NO:42; encoded by GENBANK® ID AAV70918.1), and the Bacillus sphaericus MTX2 protein (SEQ ID NO:16, GENBANK® ID AAC44124.1).
Example 7
Axmi-011, axmi-012 and axmi-015
[0115] From strain ATX13008, three individual clones were found to contain open reading frames with homology to MTX-like toxins. These open reading frames were designated axmi-011 (SEQ ID NO:1), axmi-012 (SEQ ID NO:3), and axmi-015 (SEQ ID NO:8), and the encoded proteins are designated AXMI-011 (SEQ ID NO:2), AXMI-012 (SEQ ID NO:4), and AXMI-015 (SEQ ID NO:9), respectively. By searching of public databases of protein sequences, AXMI-011 was found to have low homology to a class of toxins including MTX2 (SEQ ID NO: 16); AXMI-015 was found to have low homology (about 35% amino acid identify over 178 amino acids) to a mosquitocidal toxin from Bacillus thuringiensis israelensis RBTH--02046 (SEQ ID NO:41, GENBANK® ID gi|75761628:1-79; AXMI-012 was found to have homology (29% over 217 amino acids) to a class of toxins including the p42 binary toxin of Bacillus sphaericus (SEQ ID NO:39; GENBANK® ID CAA73761).
[0116] Inspection of the axmi-011 coding region reveals the existence of an alternate translational start site 12 nucleotides upstream of the ATG start of axmi-011. This open reading frame contains a 5' extension of the following twelve nucleotides
TABLE-US-00002 (SEQ ID NO: 59) 5'-G-T-G-A-T-G-A-A-A-A-A-A-3'
immediately upstream and adjacent to the axmi-11 open reading frame. This open reading frame is herein designated as axmi-011(long) (SEQ ID NO:60). Translation of axmi-011 utilizing the putative GTG start would result in a modified AXMI-011 protein that contains an N-terminal extension of four amino acids (amino acid residues 1 through 4 of SEQ ID NO:61).
[0117] Analysis of the DNA context surrounding the two potential start sites reveals a sequence with a good match to the consensus for a ribosome binding site 5' G-T-G-A-T-G-3' (SEQ ID NO:62) positioned from -10 to -6 nt relative to the ATG start codon of SEQ ID NO: 1. This is a proper position for a bacterial ribosome binding site. No obvious homology to the consensus ribosome start site is observed in the position 15 nt upstream of the putative GTG start site. Thus, the protein initiated from the ATG start codon is designated AXMI-011 (SEQ ID NO:2). The protein encoded by translation initiated at the GTG start codon is designated AXMI-011(LONG) (SEQ ID NO:61).
Example 8
Axmi-032
[0118] From strain ATX9387, a plasmid was found to contain an open reading frame with homology to pesticidal toxins. This open reading frame was designated as axmi-032 (SEQ ID NO: 12), and the encoded protein was designated AXMI-032 (SEQ ID NO: 13). By searching of public databases of protein sequences, such as the GenPept database, AXMI-032 was found to have homology to a class of toxins including a presumed binary toxin from Bacillus thuringiensis (SEQ ID NO:43; GENBANK® Accession No. CAD30104.1) which is a possible two-domain toxin from Bacillus thuringiensis serovar israelensis.
Example 9
Axmi-013
[0119] From strain ATX13002, a clone was found to contain an open reading frame with homology to "cry" type delta-endotoxins. This open reading frame was designated as axmi-013 (SEQ ID NO:5), and the encoded protein was designated AXMI-013 (SEQ ID NO:6). By searching of public databases of protein sequences, the C-terminal region of AXMI-013 was found to have 52% identity with the MTX3 toxin (SEQ ID NO:40: GENBANK® ID AAB36661).
Example 10
Expression of AXMI-013 in Bacillus
[0120] The insecticidal AXMI-013 gene is amplified by PCR and cloned into the Bacillus expression vector pAX916 by methods well known in the art. The resulting clone is assayed for expression of AXMI-013 protein after transformation into cells of a cry(-) Bacillus thuringiensis strain. A Bacillus strain containing the axmi-013 clone and expressing the AXMI-013 insecticidal protein is grown in, for example, CYS media (10 g/l Bacto-casitone; 3 g/l yeast extract; 6 g/l KH2PO4; 14 g/l K2HPO4; 0.5 mM MgSO4; 0.05 mM MnCl2; 0.05 mM FeSO4), until sporulation is evident by microscopic examination. Samples are prepared, and analyzed by polyacrylamide gel electrophoresis (PAGE). AXMI-013 is tested for insecticidal activity in bioassays against important insect pests.
[0121] Inspection of the predicted amino acid sequence of AXMI-013 (SEQ ID NO:6) suggested that the N-terminus of the full-length AXMI-013 protein may comprise a signal peptide for secretion. The predicted site of cleavage was estimated to be between the alanine at position 27 and lysine at position 28 of SEQ ID NO:6. Similarly, MTX3 (SEQ ID NO:40) is predicted to possess a secretion signal peptide at its N-terminus (Liu, et al. (1996) Appl. Environ. Microbiol. 62:2174-2176).
[0122] The expressed AXMI-013 protein was excised from a polyacrylamide gel and subjected to N-terminal sequence analysis as known in the art. The N-terminal sequence identified by this analysis corresponded to a N-terminal truncation of the AXMI-013 protein, resulting in a truncated peptide with an N-terminus corresponding to the glutamine (Q) at amino acid position 40 of SEQ ID NO:6. This truncated protein is referred to herein as AXMI-013(Q), and the amino acid sequence of this protein is provided in SEQ ID NO:7. As known in the art, prediction of the exact site of cleavage is somewhat difficult. Nonetheless, the cleavage at approximately amino acid position 40 of SEQ ID NO:6 suggests that either (1) AXMI-013 is further processed after initial cleavage at amino acid positions 27/28, or AXMI-013 has a novel secretion signal. In order to confirm this, one skilled in the art may make gene fusion constructs utilizing (1) a heterologous protein and (2) using increasing length portions of AXMI-013. One can then test for secretion of the marker protein and determine the processing sites by N-terminal sequencing. Other methods to determine the extent of the signal sequence are known in the art.
Example 11
Axmi-023 and axmi-041
[0123] From strain ATX13045, a plasmid clone was found to contain an open reading frame with homology to "cry" type delta-endotoxins. This open reading frame was designated as axmi-023 (SEQ ID NO:30), and the encoded protein was designated AXMI-023 (SEQ ID NO:31). BLAST search of the non-redundant `nr` database demonstrates that AXMI-023 has low amino acid identity (less than 30% amino acid identity) with the VIP2 protein toxin, as well as several other presumed or known toxins. (GENBANK® Accession No. AAO86513.1, SEQ ID NO:55)
[0124] From strain ATX21738, a plasmid clone was found to contain an open reading frame with homology to "cry" type delta-endotoxins. This open reading frame was designated as axmi-041 (SEQ ID NO:32), and the encoded protein was designated AXMI-041 (SEQ ID NO:33). pAX4310 was deposited with the ARS Patent Strain Collection on Jun. 15, 2006, and assigned NRRL B-30943. AXMI-041 is 21% identical to AXMI-023, and similarly exhibits low amino acid identity (less than 30% amino acid identity) with the VIP2 protein toxin, as well as several other presumed or known toxins. A search of DNA and protein databases with the DNA sequences and amino acid sequences of AXMI-023 and AXMI-041 revealed that they are homologous to known pesticidal proteins. FIG. 4 shows an alignment of AXMI-023 with the Vip2 pesticidal protein (SEQ ID NO:55), and several related toxins. AXMI-041 also shows homology with this class of toxins.
Example 12
AXMI-022 Defines a Novel Class of Pesticidal Proteins
[0125] From strain ATX13045, a plasmid clone was found to contain an open reading frame with homology to known insect toxins. This open reading frame was designated as axmi-022 (SEQ ID NO:28), and the encoded protein was designated AXMI-022 (SEQ ID NO:29).
[0126] The amino acid sequence of AXMI-022 is 64.9% identical to Vip1A(b) (SEQ ID NO:52; see also U.S. Pat. No. 5,770,696, herein incorporated by reference in its entirety) throughout the length of Vip1A(b), and has significant amino acid identity with several other binary protein toxins. Further analysis of AXMI-022 revealed the following features of this polypeptide:
[0127] AXMI-022 is significantly longer than other binary proteins to which it shares homology, and encodes a peptide of 1,003 amino acids. For example, the Vip1A(b) protein is 834 amino acids in length;
[0128] Inspection of the DNA region surrounding the axmi-022 open reading frame shows no evidence for a second ORF encoding a toxin domain. The genes encoding binary toxins are typically physically closely linked. Most often, both the toxin and the receptor protein are organized as adjacent open reading frames, and are often transcriptionally linked in an operon.
[0129] The receptor binding region of AXMI-022 (from about amino acid 640 to about amino acid 770 of SEQ ID NO:29) is different from other binary toxins. The region of AXMI-022 corresponding to the region of binary proteins involved in receptor binding is quite different in AXMI-022 compared to other binary proteins. This is suggestive that AXMI-022 binds to a different receptor than other binary proteins
[0130] The C-terminal 133 amino acids of AXMI-022 (starting about amino acid 870 of SEQ ID NO:29) show amino acid homology to the Cry37Aa binary protein (SEQ ID NO:54; GENBANK® Accession No. AAF76376; U.S. Pat. No. 6,063,756, herein incorporated by reference in its entirety). This region has homology to Cry37Aa at a sequence identity of about 36%. Cry37Aa is in a different class of binary toxins than the Vip1-type toxins. Cry37Aa belongs to the Cry34 family, which forms a binary toxin with the Cry35 family. Cry34Ab1 is principally responsible for forming pores in lipid membranes, while Cry35Ab1 enhances the formation of pores (Masson, et al. (2004) Biochemistry 43:12349-57).
[0131] Thus, AXMI-022 appears to be a novel type of "single peptide" binary toxin, having homology to multiple classes of binary proteins, with a receptor-binding component from one class of binary toxins directly fused to a toxin component from a different class of binary toxins. This organization of domains has not been previously predicted in the art.
Example 13
Axmi-043
[0132] From strain ATX15398, pAX2597 was found to contain an open reading frame with homology to "cry" type delta-endotoxins. This open reading frame was designated as axmi-043. pAX2597 was deposited with the ARS Patent Strain Collection on Jun. 15, 2006, and assigned NRRL B-30941. Inspection of the axmi-043 open reading frame suggests that more than one start codon may be present. The ATG at position 46 of SEQ ID NO:20 has a ribosome binding site (5' G-G-A-G-A-3') (SEQ ID NO:63) starting at nucleotide 33 of SEQ ID NO:20. Based on the presence of this strong ribosome binding consensus sequence immediately upstream of this second start site, and the homology of the two predicted proteins to other endotoxins, we herein designate the translation product of the internal start site (represented by SEQ ID NO:22) as AXMI-043 (SEQ ID NO:23), and the longer protein as AXMI-043-2 (SEQ ID NO:21). AXMI-043 exhibits 93% amino acid identity to the AXMI-028 endotoxin, and AXMI-43-2 is 90% identical to AXMI-028 (SEQ ID NO:45 of this application, see also U.S. patent application Ser. No. 11/416,261, herein incorporated by reference in its entirety). AXMI-043 appears to be a "full-length endotoxin," and contains a C-terminal region (after the aspartic residue at position 629 of the AXMI-043 amino acid sequence) often referred to in the art as a non-toxic domain or a "crystal domain." AXMI-043 exhibits 54% amino acid identity with the Cry7Aa1 endotoxin (SEQ ID NO:46) throughout the full length of the protein.
Example 14
Axmi-44
[0133] From strain ATX14759, pAX2599 was found to contain an open reading frame with homology to "cry" type delta-endotoxins. This open reading frame was designated as axmi-044 (SEQ ID NO: 14), and the encoded protein was designated AXMI-044 (SEQ ID NO: 15). pAX2599 was deposited with the ARS Patent Strain Collection on Jun. 15, 2006, and assigned Accession No. NRRL B-30942. By searching of public databases of protein sequences, such as the GenPept database maintained by the NCBI (National Center for Biotechnology Information) AXMI-044 was found to have low homology to the cry-15Aa/cry33 family of toxins (SEQ ID NO:44, GENBANK® ID 8928022), and to MTX2 (SEQ ID NO:38).
Example 15
Axmi-033 and axmi-034
[0134] From strain ATX14833, a plasmid clone was found to contain two open reading frames with homology to insect toxins. The first open reading frame was designated as axmi-033 (SEQ ID NO:24), and the encoded protein was designated AXMI-033 (SEQ ID NO:25). AXMI-33 exhibits 61% amino acid identity to the 326 amino acid CryC35 insect toxin (SEQ ID NO:47, encoded by GENBANK® reference CAA63374). The second open reading frame was designated as axmi-034 (SEQ ID NO:26), and the encoded protein was designated AXMI-034 (SEQ ID NO:27). AXMI-34 exhibits 45% amino acid identity with the CryC53 endotoxin. (SEQ ID NO:48, encoded by GENBANK® reference CAA67205).
[0135] axmi-033 and axmi-034 appear to comprise an operon. The ATG start of axmi-034 is immediately 3' to, and in close proximity of (15 nucleotides immediately downstream of), the TAA stop codon of axmi-033. This organization is well known in the art to suggest that two genes comprise an operon. Thus, the AXMI-033 and AXMI-034 proteins are likely to be co-expressed in their native strain. It is likely that the activities of the two proteins expressed together may be synergistic and superior to the activity of the proteins expressed separately. pAX4341, a clone containing both axmi-033 and axmi-034 open reading frames, was deposited with the ARS Patent Strain Collection on May 29, 2007, and assigned accession number NRRL B-50047.
Example 16
Axmi-063 and axmi-064
[0136] From strain ATX12972, a plasmid clone was found to contain two open reading frames with homology to insect toxins. The first open reading frame was designated as axmi-063 (SEQ ID NO:34), and the encoded protein was designated AXMI-063 (SEQ ID NO:35). AXMI-63 exhibits 53.1% amino acid identity to the CryC35 insect toxin (SEQ ID NO:47, encoded by GENBANK® reference CAA63374). The second open reading frame was designated as axmi-064 (SEQ ID NO:36), and the encoded protein was designated AXMI-064 (SEQ ID NO:37). AXMI-64 exhibits 44.3% amino acid identity with the CryC53 endotoxin (SEQ ID NO:48, encoded by GENBANK® reference CAA67205).
[0137] axmi-063 and axmi-064 appear to comprise an operon. The ATG start of axmi-064 is immediately 3' to, and in close proximity of, the TAA stop codon of axmi-063. This is an organization well known in the art to suggest that two genes comprise an operon. Thus, the AXMI-063 and AXMI-064 proteins are likely to be co-expressed in their native strain. It is likely that the activities of the two proteins expressed together may be synergistic and superior to the activity of the proteins expressed separately. pAX5036, a clone containing both axmi-063 and axmi-064 open reading frames, was deposited with the ARS Patent Strain Collection on May 29, 2007, and assigned NRRL B-50048.
[0138] AXMI-033/AXMI-034 are similar to AXMI-063/AXMI-064. Analysis of the amino acid sequence of AXMI-033, AXMI-043, AXMI-063, and AXMI-064 reveals that AXMI-033 and AXMI-063 share significant amino acid identity, and are 69% identical. Similarly AXMI-034 and AXMI-064 share significant amino acid similarity (52% identical).
Example 17
Homology of AXMI-011, AXMI-012, AXMI-013, AXMI-015, AXMI-032, and AXMI-044 to Known Pesticidal Protein Genes
[0139] A search of protein databases with the amino acid sequences of the proteins of the invention reveal that they are homologous to known pesticidal proteins. Comparison of the amino acid sequences of the proteins of the invention to the non-redundant (nr) database maintained by the NCBI using the BLAST algorithm revealed the following proteins as having the strongest block of amino acid identity to the sequences of the invention (Table 2). Thus, the proteins of the invention are "pesticidal proteins."
TABLE-US-00003 TABLE 2 Amino Acid Identity of AXMI-011, AXMI-012, AXMI-013, AXMI-015, AXMI-032, and AXMI-044 to mosquito toxins in public databases % Identity in PROTEIN Highest Blast Hit (nr) block AXMI-011 MTX2 28% AXMI-012 P42 component of binary toxin 29% AXMI-013 MTX3 52% AXMI-015 RBTH_02046 35% AXMI-019 Cry14-4, MTX2 35%, 30% AXMI-032 GENBANK ® ID CAD30104.1 19% AXMI-044 cry15Aa, MTX2 30%, 30%
Example 18
Additional Assays for Pesticidal Activity
[0140] The ability of a pesticidal protein to act as a pesticide upon a pest is often assessed in a number of ways. One way well known in the art is to perform a feeding assay. In such a feeding assay, one exposes the pest to a sample containing either compounds to be tested, or control samples. Often this is performed by placing the material to be tested, or a suitable dilution of such material, onto a material that the pest will ingest, such as an artificial diet. The material to be tested may be composed of a liquid, solid, or slurry. The material to be tested may be placed upon the surface and then allowed to dry. Alternatively, the material to be tested may be mixed with a molten artificial diet, then dispensed into the assay chamber. The assay chamber may be, for example, a cup, a dish, or a well of a microtiter plate.
[0141] Assays for sucking pests (for example aphids) may involve separating the test material from the insect by a partition, ideally a portion that can be pierced by the sucking mouth parts of the sucking insect, to allow ingestion of the test material. Often the test material is mixed with a feeding stimulant, such as sucrose, to promote ingestion of the test compound.
[0142] Other types of assays can include microinjection of the test material into the mouth, or gut of the pest, as well as development of transgenic plants, followed by test of the ability of the pest to feed upon the transgenic plant. Plant testing may involve isolation of the plant parts normally consumed, for example, small cages attached to a leaf, or isolation of entire plants in cages containing insects.
[0143] Other methods and approaches to assay pests are known in the art, and can be found, for example in Robertson and Preisler, eds. (1992) Pesticide bioassays with arthropods, CRC, Boca Raton, Fla. Alternatively, assays are commonly described in the journals Arthropod Management Tests and Journal of Economic Entomology or by discussion with members of the Entomological Society of America (ESA).
Example 19
Vectoring of Axmi Genes for Plant Expression
[0144] The coding regions of the invention are connected with appropriate promoter and terminator sequences for expression in plants. Such sequences are well known in the art and may include the rice actin promoter or maize ubiquitin promoter for expression in monocots, the Arabidopsis UBQ3 promoter or CaMV 35S promoter for expression in dicots, and the nos or PinII terminators. Techniques for producing and confirming promoter--gene--terminator constructs also are well known in the art.
[0145] In one aspect of the invention, synthetic DNA sequences are designed and generated. These synthetic sequences have altered nucleotide sequence relative to the parent sequence, but encode proteins that are essentially identical to the parent AXMI protein.
[0146] In another aspect of the invention, modified versions of the synthetic genes are designed such that the resulting peptide is targeted to a plant organelle, such as the endoplasmic reticulum or the apoplast. Peptide sequences known to result in targeting of fusion proteins to plant organelles are known in the art. For example, the N-terminal region of the acid phosphatase gene from the White Lupin Lupinus albus (GENEBANK® ID GI:14276838, Miller et al. (2001) Plant Physiology 127: 594-606) is known in the art to result in endoplasmic reticulum targeting of heterologous proteins. If the resulting fusion protein also contains an endoplasmic reticulum retention sequence comprising the peptide N-terminus-lysine-aspartic acid-glutamic acid-leucine (i.e., the "KDEL" motif (SEQ ID NO:58)) at the C-terminus, the fusion protein will be targeted to the endoplasmic reticulum. If the fusion protein lacks an endoplasmic reticulum targeting sequence at the C-terminus, the protein will be targeted to the endoplasmic reticulum, but will ultimately be sequestered in the apoplast.
[0147] Thus, this gene encodes a fusion protein that contains the N-terminal thirty-one amino acids of the acid phosphatase gene from the White Lupin Lupinus albus (GENBANK® ID GI: 14276838, Miller et al., 2001, supra) fused to the N-terminus of the AXMI sequence, as well as the KDEL sequence at the C-terminus. Thus, the resulting protein is predicted to be targeted the plant endoplasmic reticulum upon expression in a plant cell.
[0148] The plant expression cassettes described above are combined with an appropriate plant selectable marker to aid in the selection of transformed cells and tissues, and ligated into plant transformation vectors. These may include binary vectors from Agrobacterium-mediated transformation or simple plasmid vectors for aerosol or biolistic transformation.
Example 20
Vectoring of Axmi Genes for Plant Expression
[0149] The coding region DNA of the axmi genes of the invention are operably connected with appropriate promoter and terminator sequences for expression in plants. Such sequences are well known in the art and may include the rice actin promoter or maize ubiquitin promoter for expression in monocots, the Arabidopsis UBQ3 promoter or CaMV 35S promoter for expression in dicots, and the nos or PinII terminators. Techniques for producing and confirming promoter--gene--terminator constructs also are well known in the art.
[0150] The plant expression cassettes described above are combined with an appropriate plant selectable marker to aid in the selections of transformed cells and tissues, and ligated into plant transformation vectors. These may include binary vectors from Agrobacterium-mediated transformation or simple plasmid vectors for aerosol or biolistic transformation.
Example 21
Transformation of Maize Cells with the Pesticidal Protein Genes Described Herein
[0151] Maize ears are best collected 8-12 days after pollination. Embryos are isolated from the ears, and those embryos 0.8-1.5 mm in size are preferred for use in transformation. Embryos are plated scutellum side-up on a suitable incubation media, such as DN62A5S media (3.98 g/L N6 Salts; 1 mL/L (of 1000× Stock) N6 Vitamins; 800 mg/L L-Asparagine; 100 mg/L Myo-inositol; 1.4 g/L L-Proline; 100 mg/L Casamino acids; 50 g/L sucrose; 1 mL/L (of 1 mg/mL Stock) 2,4-D). However, media and salts other than DN62A5S are suitable and are known in the art. Embryos are incubated overnight at 25° C. in the dark. However, it is not necessary per se to incubate the embryos overnight.
[0152] The resulting explants are transferred to mesh squares (30-40 per plate), transferred onto osmotic media for about 30-45 minutes, then transferred to a beaming plate (see, for example, PCT Publication No. WO/0138514 and U.S. Pat. No. 5,240,842).
[0153] DNA constructs designed to the genes of the invention in plant cells are accelerated into plant tissue using an aerosol beam accelerator, using conditions essentially as described in PCT Publication No. WO/0138514. After beaming, embryos are incubated for about 30 min on osmotic media, and placed onto incubation media overnight at 25° C. in the dark. To avoid unduly damaging beamed explants, they are incubated for at least 24 hours prior to transfer to recovery media. Embryos are then spread onto recovery period media, for about 5 days, 25° C. in the dark, then transferred to a selection media. Explants are incubated in selection media for up to eight weeks, depending on the nature and characteristics of the particular selection utilized. After the selection period, the resulting callus is transferred to embryo maturation media, until the formation of mature somatic embryos is observed. The resulting mature somatic embryos are then placed under low light, and the process of regeneration is initiated by methods known in the art. The resulting shoots are allowed to root on rooting media, and the resulting plants are transferred to nursery pots and propagated as transgenic plants.
TABLE-US-00004 Materials DN62A5S Media Components Per Liter Source Chu's N6 Basal Salt Mixture 3.98 g/L Phytotechnology Labs (Prod. No. C 416) Chu's N6 Vitamin Solution 1 mL/L Phytotechnology Labs (Prod. No. C 149) (of 1000x Stock) L-Asparagine 800 mg/L Phytotechnology Labs Myo-inositol 100 mg/L Sigma L-Proline 1.4 g/L Phytotechnology Labs Casamino acids 100 mg/L Fisher Scientific Sucrose 50 g/L Phytotechnology Labs 2,4-D (Prod. No. D-7299) 1 mL/L Sigma (of 1 mg/mL Stock)
[0154] The pH of the solution is adjusted to pH 5.8 with 1N KOH/1N KCl, Gelrite (Sigma) is added at a concentration up to 3 g/L, and the media is autoclaved. After cooling to 50° C., 2 ml/L of a 5 mg/ml stock solution of silver nitrate (Phytotechnology Labs) is added.
Example 22
Transformation of the Pesticidal Genes of the Invention in Plant Cells by Agrobacterium-Mediated Transformation
[0155] Ears are best collected 8-12 days after pollination. Embryos are isolated from the ears, and those embryos 0.8-1.5 mm in size are preferred for use in transformation. Embryos are plated scutellum side-up on a suitable incubation media, and incubated overnight at 25° C. in the dark. However, it is not necessary per se to incubate the embryos overnight. Embryos are contacted with an Agrobacterium strain containing the appropriate vectors for Ti plasmid mediated transfer for about 5-10 min, and then plated onto co-cultivation media for about 3 days (25° C. in the dark). After co-cultivation, explants are transferred to recovery period media for about five days (at 25° C. in the dark). Explants are incubated in selection media for up to eight weeks, depending on the nature and characteristics of the particular selection utilized. After the selection period, the resulting callus is transferred to embryo maturation media, until the formation of mature somatic embryos is observed. The resulting mature somatic embryos are then placed under low light, and the process of regeneration is initiated as known in the art.
Example 23
Soil Infestation of Plants Expressing a Gene of the Invention with Western Corn Rootworm
[0156] Transgenic plants containing an axmi gene of the invention under the control of a plant promoter are tested for resistance to infestation by Western corn rootworm (WCRW). Plantlets are transplanted from tissue culture media to root trainer (clamshell) pots known in the art to be useful for growth of plantlets in soil. Plants are grown for about 2 weeks in a greenhouse. Transgenic plants, as well as untransformed controls, are infested with approximately 1,000 WCRW eggs. WCRW eggs are preincubated such that eggs are at the point of hatching when infested onto the plants. Plants are held for about four weeks, or until controls exhibited obvious damage due to the rootworms. At this stage, plants are pulled from pots, roots are washed, and damage evaluated. Several independent events are examined for reduced damage from WCRW infestation relative to non-transformed control plants.
[0157] The resulting shoots are allowed to root on rooting media, and the resulting plants are transferred to nursery pots and propagated as transgenic plants.
[0158] All publications and patent applications mentioned in the specification are indicative of the level of skill of those skilled in the art to which this invention pertains. All publications and patent applications are herein incorporated by reference to the same extent as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference.
[0159] Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, it will be obvious that certain changes and modifications may be practiced within the scope of the appended claims.
Sequence CWU
1
1
631957DNABacillus thuringiensisCDS(1)...(957) 1atg aat aaa aaa cct atg gta
gcg ttg ata tta gcc act tcg att ggt 48Met Asn Lys Lys Pro Met Val
Ala Leu Ile Leu Ala Thr Ser Ile Gly1 5 10
15ata cct tgt aca ttt aca cct gga agt gca tta gca gca
gaa aat att 96Ile Pro Cys Thr Phe Thr Pro Gly Ser Ala Leu Ala Ala
Glu Asn Ile 20 25 30cag act
agt gtt aat gaa aat gta aaa gtt ggt att aca gat gtt caa 144Gln Thr
Ser Val Asn Glu Asn Val Lys Val Gly Ile Thr Asp Val Gln 35
40 45tct gaa ttg aat aag ata gga gac tat tat
tat agt aat aac tta gca 192Ser Glu Leu Asn Lys Ile Gly Asp Tyr Tyr
Tyr Ser Asn Asn Leu Ala 50 55 60aat
acg act ata aaa cct cct cat cat tgg gat tat aca ctt aaa aaa 240Asn
Thr Thr Ile Lys Pro Pro His His Trp Asp Tyr Thr Leu Lys Lys65
70 75 80aat cct gat aaa gtt gga
aca aat ttg gat ttt agt att act ggt act 288Asn Pro Asp Lys Val Gly
Thr Asn Leu Asp Phe Ser Ile Thr Gly Thr 85
90 95gct agt aaa cta aat tat gat agt gta act cct ata
tac att ggg cat 336Ala Ser Lys Leu Asn Tyr Asp Ser Val Thr Pro Ile
Tyr Ile Gly His 100 105 110aat
gaa ttt aat aat gat tca gat cag cct caa aaa ttt aca act tct 384Asn
Glu Phe Asn Asn Asp Ser Asp Gln Pro Gln Lys Phe Thr Thr Ser 115
120 125aaa ttt act aaa gct gta aca gag gga
aca aca agt acc gta aca aat 432Lys Phe Thr Lys Ala Val Thr Glu Gly
Thr Thr Ser Thr Val Thr Asn 130 135
140gga ttt aga tta gga aat cca ggt tta aac tta ttt act att cca tta
480Gly Phe Arg Leu Gly Asn Pro Gly Leu Asn Leu Phe Thr Ile Pro Leu145
150 155 160att tta agt gat
ggt atg aaa att aat gcg gaa ttt aac tct tct act 528Ile Leu Ser Asp
Gly Met Lys Ile Asn Ala Glu Phe Asn Ser Ser Thr 165
170 175tca gaa tct caa caa aaa tcg gaa aca aaa
aca ata gaa gca tca cct 576Ser Glu Ser Gln Gln Lys Ser Glu Thr Lys
Thr Ile Glu Ala Ser Pro 180 185
190caa aac ata gaa gtt cca gca cat aaa aaa tat aaa gta gat gtt gta
624Gln Asn Ile Glu Val Pro Ala His Lys Lys Tyr Lys Val Asp Val Val
195 200 205ttg gaa caa aca agc tat tgg
gca gat gtt aca ttt aca ggt gaa gga 672Leu Glu Gln Thr Ser Tyr Trp
Ala Asp Val Thr Phe Thr Gly Glu Gly 210 215
220att aat ctt aat act act ata aat gca act gga ata cat act ggg cat
720Ile Asn Leu Asn Thr Thr Ile Asn Ala Thr Gly Ile His Thr Gly His225
230 235 240atg gga atg cag
gag tca aga aaa ttt tct tgg aac aaa aat acc att 768Met Gly Met Gln
Glu Ser Arg Lys Phe Ser Trp Asn Lys Asn Thr Ile 245
250 255gaa tta ttt aat gga cta aaa caa gag caa
aaa aat aat ata cat ggg 816Glu Leu Phe Asn Gly Leu Lys Gln Glu Gln
Lys Asn Asn Ile His Gly 260 265
270att aaa ttt agt aat ggg aaa atg aat gca aac gga aca ggt aaa gtt
864Ile Lys Phe Ser Asn Gly Lys Met Asn Ala Asn Gly Thr Gly Lys Val
275 280 285gaa ggt att ttt ggt agt aat
cta gtt gta aag gta aat gat gtt aca 912Glu Gly Ile Phe Gly Ser Asn
Leu Val Val Lys Val Asn Asp Val Thr 290 295
300gat cca tta aat cct atc cta gta atg act aaa agt tta aaa taa
957Asp Pro Leu Asn Pro Ile Leu Val Met Thr Lys Ser Leu Lys 305
310 3152318PRTBacillus thuringiensis 2Met Asn
Lys Lys Pro Met Val Ala Leu Ile Leu Ala Thr Ser Ile Gly1 5
10 15 Ile Pro Cys Thr Phe Thr Pro
Gly Ser Ala Leu Ala Ala Glu Asn Ile 20 25
30 Gln Thr Ser Val Asn Glu Asn Val Lys Val Gly Ile
Thr Asp Val Gln 35 40 45
Ser Glu Leu Asn Lys Ile Gly Asp Tyr Tyr Tyr Ser Asn Asn Leu Ala
50 55 60 Asn Thr Thr
Ile Lys Pro Pro His His Trp Asp Tyr Thr Leu Lys Lys65 70
75 80 Asn Pro Asp Lys Val Gly Thr Asn
Leu Asp Phe Ser Ile Thr Gly Thr 85 90
95 Ala Ser Lys Leu Asn Tyr Asp Ser Val Thr Pro Ile Tyr
Ile Gly His 100 105 110
Asn Glu Phe Asn Asn Asp Ser Asp Gln Pro Gln Lys Phe Thr Thr Ser
115 120 125 Lys Phe Thr Lys
Ala Val Thr Glu Gly Thr Thr Ser Thr Val Thr Asn 130
135 140 Gly Phe Arg Leu Gly Asn Pro Gly
Leu Asn Leu Phe Thr Ile Pro Leu145 150
155 160 Ile Leu Ser Asp Gly Met Lys Ile Asn Ala Glu Phe
Asn Ser Ser Thr 165 170
175 Ser Glu Ser Gln Gln Lys Ser Glu Thr Lys Thr Ile Glu Ala Ser Pro
180 185 190 Gln Asn Ile
Glu Val Pro Ala His Lys Lys Tyr Lys Val Asp Val Val 195
200 205 Leu Glu Gln Thr Ser Tyr Trp Ala
Asp Val Thr Phe Thr Gly Glu Gly 210 215
220 Ile Asn Leu Asn Thr Thr Ile Asn Ala Thr Gly Ile His
Thr Gly His225 230 235
240 Met Gly Met Gln Glu Ser Arg Lys Phe Ser Trp Asn Lys Asn Thr Ile
245 250 255 Glu Leu Phe Asn
Gly Leu Lys Gln Glu Gln Lys Asn Asn Ile His Gly 260
265 270 Ile Lys Phe Ser Asn Gly Lys Met Asn
Ala Asn Gly Thr Gly Lys Val 275 280
285 Glu Gly Ile Phe Gly Ser Asn Leu Val Val Lys Val Asn Asp
Val Thr 290 295 300
Asp Pro Leu Asn Pro Ile Leu Val Met Thr Lys Ser Leu Lys305
310 315 3927DNABacillus
thuringiensisCDS(1)...(927) 3atg att aat gta aac agt ggt aag gtt ata gat
ata tca gga aat tca 48Met Ile Asn Val Asn Ser Gly Lys Val Ile Asp
Ile Ser Gly Asn Ser1 5 10
15aca gca aat att caa caa tat gag tgg cgt ggt gat tta cca tct gaa
96Thr Ala Asn Ile Gln Gln Tyr Glu Trp Arg Gly Asp Leu Pro Ser Glu
20 25 30tat tgg tac ttt cat cgt gaa
gcg gat ggg tat tat gtt att gaa tca 144Tyr Trp Tyr Phe His Arg Glu
Ala Asp Gly Tyr Tyr Val Ile Glu Ser 35 40
45aaa ctt agt gga aag gta tta gac ata aaa gga aat tca aca gca
aat 192Lys Leu Ser Gly Lys Val Leu Asp Ile Lys Gly Asn Ser Thr Ala
Asn 50 55 60aat gca aat gtt cag caa
tat caa ttt ata caa aat gca cct gct gaa 240Asn Ala Asn Val Gln Gln
Tyr Gln Phe Ile Gln Asn Ala Pro Ala Glu65 70
75 80aga ttt gct gct gaa gaa gcg gga agc gtc tct
ctc cca tca ata aat 288Arg Phe Ala Ala Glu Glu Ala Gly Ser Val Ser
Leu Pro Ser Ile Asn 85 90
95acg aaa cca tta tca cca gta cca gaa tat aaa acg att aat gat caa
336Thr Lys Pro Leu Ser Pro Val Pro Glu Tyr Lys Thr Ile Asn Asp Gln
100 105 110ctt cca gaa gaa acg gaa
cgt gta gta aca gct ttt aca ata gtt ccg 384Leu Pro Glu Glu Thr Glu
Arg Val Val Thr Ala Phe Thr Ile Val Pro 115 120
125tgt atc tca gta aaa gat cca cat tat ggt gga gat act gct
aaa caa 432Cys Ile Ser Val Lys Asp Pro His Tyr Gly Gly Asp Thr Ala
Lys Gln 130 135 140ata aga gaa aat cct
tat tac atg gtt gta aaa aaa caa tgg tgg aaa 480Ile Arg Glu Asn Pro
Tyr Tyr Met Val Val Lys Lys Gln Trp Trp Lys145 150
155 160aaa caa gaa tct tat gtt tta gct cct agt
gaa acg tat act ttt gaa 528Lys Gln Glu Ser Tyr Val Leu Ala Pro Ser
Glu Thr Tyr Thr Phe Glu 165 170
175aca aaa act ggt ata aaa gta att gat caa gaa act gct aca agg aca
576Thr Lys Thr Gly Ile Lys Val Ile Asp Gln Glu Thr Ala Thr Arg Thr
180 185 190gta agt tgg agc att ggt
gct gat atg gga ttt agt ttc aaa gga ttt 624Val Ser Trp Ser Ile Gly
Ala Asp Met Gly Phe Ser Phe Lys Gly Phe 195 200
205tca tta gga atg tcc act caa tat tca aca caa tta cag act
act ata 672Ser Leu Gly Met Ser Thr Gln Tyr Ser Thr Gln Leu Gln Thr
Thr Ile 210 215 220agt cat aca act gaa
caa tta aaa gaa gaa aca aac agg cac gaa ata 720Ser His Thr Thr Glu
Gln Leu Lys Glu Glu Thr Asn Arg His Glu Ile225 230
235 240aag aat cca ttt tca gag aga atg gcg tat
tct aga tat gta tta gcg 768Lys Asn Pro Phe Ser Glu Arg Met Ala Tyr
Ser Arg Tyr Val Leu Ala 245 250
255aca gaa tat tct gtc caa aga aaa aat ggt aca atc gta aat tct cct
816Thr Glu Tyr Ser Val Gln Arg Lys Asn Gly Thr Ile Val Asn Ser Pro
260 265 270tgg act atg acc gat aag
aca aaa gca cat gct gta act ttt cca aaa 864Trp Thr Met Thr Asp Lys
Thr Lys Ala His Ala Val Thr Phe Pro Lys 275 280
285tcc aca gga aat gca tta gat gaa aat aca aag gaa cta tca
aat agt 912Ser Thr Gly Asn Ala Leu Asp Glu Asn Thr Lys Glu Leu Ser
Asn Ser 290 295 300gaa agt gta aac taa
927Glu Ser Val Asn
3054308PRTBacillus thuringiensis 4Met Ile Asn Val Asn Ser Gly Lys Val Ile
Asp Ile Ser Gly Asn Ser1 5 10
15 Thr Ala Asn Ile Gln Gln Tyr Glu Trp Arg Gly Asp Leu Pro Ser
Glu 20 25 30 Tyr
Trp Tyr Phe His Arg Glu Ala Asp Gly Tyr Tyr Val Ile Glu Ser 35
40 45 Lys Leu Ser Gly Lys Val
Leu Asp Ile Lys Gly Asn Ser Thr Ala Asn 50 55
60 Asn Ala Asn Val Gln Gln Tyr Gln Phe Ile Gln
Asn Ala Pro Ala Glu65 70 75
80 Arg Phe Ala Ala Glu Glu Ala Gly Ser Val Ser Leu Pro Ser Ile Asn
85 90 95 Thr Lys Pro
Leu Ser Pro Val Pro Glu Tyr Lys Thr Ile Asn Asp Gln 100
105 110 Leu Pro Glu Glu Thr Glu Arg Val
Val Thr Ala Phe Thr Ile Val Pro 115 120
125 Cys Ile Ser Val Lys Asp Pro His Tyr Gly Gly Asp Thr
Ala Lys Gln 130 135 140
Ile Arg Glu Asn Pro Tyr Tyr Met Val Val Lys Lys Gln Trp Trp Lys145
150 155 160 Lys Gln Glu Ser Tyr
Val Leu Ala Pro Ser Glu Thr Tyr Thr Phe Glu 165
170 175 Thr Lys Thr Gly Ile Lys Val Ile Asp Gln
Glu Thr Ala Thr Arg Thr 180 185
190 Val Ser Trp Ser Ile Gly Ala Asp Met Gly Phe Ser Phe Lys Gly
Phe 195 200 205 Ser
Leu Gly Met Ser Thr Gln Tyr Ser Thr Gln Leu Gln Thr Thr Ile 210
215 220 Ser His Thr Thr Glu Gln
Leu Lys Glu Glu Thr Asn Arg His Glu Ile225 230
235 240 Lys Asn Pro Phe Ser Glu Arg Met Ala Tyr Ser
Arg Tyr Val Leu Ala 245 250
255 Thr Glu Tyr Ser Val Gln Arg Lys Asn Gly Thr Ile Val Asn Ser Pro
260 265 270 Trp Thr Met
Thr Asp Lys Thr Lys Ala His Ala Val Thr Phe Pro Lys 275
280 285 Ser Thr Gly Asn Ala Leu Asp Glu
Asn Thr Lys Glu Leu Ser Asn Ser 290 295
300 Glu Ser Val Asn305 51017DNABacillus
thuringiensisCDS(1)...(1017) 5atg gat aaa aaa ata aca aaa gca gcg tta agc
atg ata atg ggt ata 48Met Asp Lys Lys Ile Thr Lys Ala Ala Leu Ser
Met Ile Met Gly Ile1 5 10
15agt gtt tta tca tct cct tta gct gta gcc gca aaa aca gag aat aat
96Ser Val Leu Ser Ser Pro Leu Ala Val Ala Ala Lys Thr Glu Asn Asn
20 25 30aaa gaa caa cac gta att aca
cag ttt aat cag aga gaa aat aag ttc 144Lys Glu Gln His Val Ile Thr
Gln Phe Asn Gln Arg Glu Asn Lys Phe 35 40
45cct gat gta gga cag ggg att caa tgg tta tct caa ttt tat gga
aaa 192Pro Asp Val Gly Gln Gly Ile Gln Trp Leu Ser Gln Phe Tyr Gly
Lys 50 55 60tct tta agg aat aat ggt
gaa gga tac tcc tta ggt cag gat gta atg 240Ser Leu Arg Asn Asn Gly
Glu Gly Tyr Ser Leu Gly Gln Asp Val Met65 70
75 80agc tat ttt cta gaa gta aaa aat tct tat ggt
caa ttg gca atg gaa 288Ser Tyr Phe Leu Glu Val Lys Asn Ser Tyr Gly
Gln Leu Ala Met Glu 85 90
95cct caa gta ata agc act aca cct ctt tgg gct ggc caa agt gac ttg
336Pro Gln Val Ile Ser Thr Thr Pro Leu Trp Ala Gly Gln Ser Asp Leu
100 105 110gaa aat gca act gat gat
gaa caa act tta aat tcc aca gaa ttt aaa 384Glu Asn Ala Thr Asp Asp
Glu Gln Thr Leu Asn Ser Thr Glu Phe Lys 115 120
125aaa acg tat tct aat aca aca acc acc tct aca gaa aat gga
ttt atg 432Lys Thr Tyr Ser Asn Thr Thr Thr Thr Ser Thr Glu Asn Gly
Phe Met 130 135 140ata ggg cag gaa acc
gaa ggg aaa gtt ggt ata ccc ttt gtc gca gaa 480Ile Gly Gln Glu Thr
Glu Gly Lys Val Gly Ile Pro Phe Val Ala Glu145 150
155 160gga aaa gtc acc ata aaa act gaa tat aat
ttt aat cat act aat ggg 528Gly Lys Val Thr Ile Lys Thr Glu Tyr Asn
Phe Asn His Thr Asn Gly 165 170
175tat gaa aca tct gag agt gta gag tat att gct cct tct caa tct att
576Tyr Glu Thr Ser Glu Ser Val Glu Tyr Ile Ala Pro Ser Gln Ser Ile
180 185 190aag gta cca ccg cat act
att gcc cga gtg aca gca tta tta gat gtg 624Lys Val Pro Pro His Thr
Ile Ala Arg Val Thr Ala Leu Leu Asp Val 195 200
205aaa aaa att aaa ggg aag atg cat cta tat tca gaa att gga
ctt aat 672Lys Lys Ile Lys Gly Lys Met His Leu Tyr Ser Glu Ile Gly
Leu Asn 210 215 220aaa gat tat ggt tac
gat atg gta cca ctt gtt tat aaa tat gga ggt 720Lys Asp Tyr Gly Tyr
Asp Met Val Pro Leu Val Tyr Lys Tyr Gly Gly225 230
235 240cca ttt aaa tat gta acc tta ggt aca tta
tat gac gag ggc tat aag 768Pro Phe Lys Tyr Val Thr Leu Gly Thr Leu
Tyr Asp Glu Gly Tyr Lys 245 250
255cag gca aaa tta gat tat tct aat atg gga aat gtt ata ccg gaa gaa
816Gln Ala Lys Leu Asp Tyr Ser Asn Met Gly Asn Val Ile Pro Glu Glu
260 265 270att gag act gtt tcg aaa
agt aac aat ccc aac cat tta tta gca agt 864Ile Glu Thr Val Ser Lys
Ser Asn Asn Pro Asn His Leu Leu Ala Ser 275 280
285gga gta gga atc ttt gaa tca gaa tac gga agt gta ttt aat
gtt aaa 912Gly Val Gly Ile Phe Glu Ser Glu Tyr Gly Ser Val Phe Asn
Val Lys 290 295 300gtt gaa tat ata gat
att aaa aat aaa aag att aaa aaa aca gag aat 960Val Glu Tyr Ile Asp
Ile Lys Asn Lys Lys Ile Lys Lys Thr Glu Asn305 310
315 320ttc act atc gaa ccg aca ata gtc cct gtt
gaa cag aag aat acg aat 1008Phe Thr Ile Glu Pro Thr Ile Val Pro Val
Glu Gln Lys Asn Thr Asn 325 330
335aca aaa taa
1017Thr Lys 6338PRTBacillus thuringiensis 6Met Asp Lys Lys Ile Thr Lys
Ala Ala Leu Ser Met Ile Met Gly Ile1 5 10
15 Ser Val Leu Ser Ser Pro Leu Ala Val Ala Ala Lys
Thr Glu Asn Asn 20 25 30
Lys Glu Gln His Val Ile Thr Gln Phe Asn Gln Arg Glu Asn Lys Phe
35 40 45 Pro Asp Val Gly
Gln Gly Ile Gln Trp Leu Ser Gln Phe Tyr Gly Lys 50 55
60 Ser Leu Arg Asn Asn Gly Glu Gly Tyr
Ser Leu Gly Gln Asp Val Met65 70 75
80 Ser Tyr Phe Leu Glu Val Lys Asn Ser Tyr Gly Gln Leu Ala
Met Glu 85 90 95
Pro Gln Val Ile Ser Thr Thr Pro Leu Trp Ala Gly Gln Ser Asp Leu
100 105 110 Glu Asn Ala Thr Asp
Asp Glu Gln Thr Leu Asn Ser Thr Glu Phe Lys 115
120 125 Lys Thr Tyr Ser Asn Thr Thr Thr Thr
Ser Thr Glu Asn Gly Phe Met 130 135
140 Ile Gly Gln Glu Thr Glu Gly Lys Val Gly Ile Pro Phe
Val Ala Glu145 150 155
160 Gly Lys Val Thr Ile Lys Thr Glu Tyr Asn Phe Asn His Thr Asn Gly
165 170 175 Tyr Glu Thr Ser
Glu Ser Val Glu Tyr Ile Ala Pro Ser Gln Ser Ile 180
185 190 Lys Val Pro Pro His Thr Ile Ala Arg
Val Thr Ala Leu Leu Asp Val 195 200
205 Lys Lys Ile Lys Gly Lys Met His Leu Tyr Ser Glu Ile Gly
Leu Asn 210 215 220
Lys Asp Tyr Gly Tyr Asp Met Val Pro Leu Val Tyr Lys Tyr Gly Gly225
230 235 240 Pro Phe Lys Tyr Val
Thr Leu Gly Thr Leu Tyr Asp Glu Gly Tyr Lys 245
250 255 Gln Ala Lys Leu Asp Tyr Ser Asn Met Gly
Asn Val Ile Pro Glu Glu 260 265
270 Ile Glu Thr Val Ser Lys Ser Asn Asn Pro Asn His Leu Leu Ala
Ser 275 280 285 Gly
Val Gly Ile Phe Glu Ser Glu Tyr Gly Ser Val Phe Asn Val Lys 290
295 300 Val Glu Tyr Ile Asp Ile
Lys Asn Lys Lys Ile Lys Lys Thr Glu Asn305 310
315 320 Phe Thr Ile Glu Pro Thr Ile Val Pro Val Glu
Gln Lys Asn Thr Asn 325 330
335 Thr Lys7296PRTBacillus thuringiensis 7Gln Arg Glu Asn Lys Phe
Pro Asp Val Gly Gln Gly Ile Gln Trp Leu1 5
10 15 Ser Gln Phe Tyr Gly Lys Ser Leu Arg Asn Asn
Gly Glu Gly Tyr Ser 20 25 30
Leu Gly Gln Asp Val Met Ser Tyr Phe Leu Glu Val Lys Asn Ser Tyr
35 40 45 Gly Gln Leu
Ala Met Glu Pro Gln Val Ile Ser Thr Thr Pro Leu Trp 50
55 60 Ala Gly Gln Ser Asp Leu Glu Asn
Ala Thr Asp Asp Glu Gln Thr Leu65 70 75
80 Asn Ser Thr Glu Phe Lys Lys Thr Tyr Ser Asn Thr Thr
Thr Thr Ser 85 90 95
Thr Glu Asn Gly Phe Met Ile Gly Gln Glu Thr Glu Gly Lys Val Gly
100 105 110 Ile Pro Phe Val Ala
Glu Gly Lys Val Thr Ile Lys Thr Glu Tyr Asn 115
120 125 Phe Asn His Thr Asn Gly Tyr Glu Thr
Ser Glu Ser Val Glu Tyr Ile 130 135
140 Ala Pro Ser Gln Ser Ile Lys Val Pro Pro His Thr Ile
Ala Arg Val145 150 155
160 Thr Ala Leu Leu Asp Val Lys Lys Ile Lys Gly Lys Met His Leu Tyr
165 170 175 Ser Glu Ile Gly
Leu Asn Lys Asp Tyr Gly Tyr Asp Met Val Pro Leu 180
185 190 Val Tyr Lys Tyr Gly Gly Pro Phe Lys
Tyr Val Thr Leu Gly Thr Leu 195 200
205 Tyr Asp Glu Gly Tyr Lys Gln Ala Lys Leu Asp Tyr Ser Asn
Met Gly 210 215 220
Asn Val Ile Pro Glu Glu Ile Glu Thr Val Ser Lys Ser Asn Asn Pro225
230 235 240 Asn His Leu Leu Ala
Ser Gly Val Gly Ile Phe Glu Ser Glu Tyr Gly 245
250 255 Ser Val Phe Asn Val Lys Val Glu Tyr Ile
Asp Ile Lys Asn Lys Lys 260 265
270 Ile Lys Lys Thr Glu Asn Phe Thr Ile Glu Pro Thr Ile Val Pro
Val 275 280 285 Glu
Gln Lys Asn Thr Asn Thr Lys 290 295
81422DNABacillus thuringiensisCDS(1)...(1422) 8atg gtt gat cgt aac ggc
atg cca aca ata gat aga agc gga ttt act 48Met Val Asp Arg Asn Gly
Met Pro Thr Ile Asp Arg Ser Gly Phe Thr1 5
10 15gtt aat cga act gtt aat tac aca aat aca aat tgg
atg gct aat att 96Val Asn Arg Thr Val Asn Tyr Thr Asn Thr Asn Trp
Met Ala Asn Ile 20 25 30gat
aat tct cgt agg atc agt gaa tta tca att cca gga aca cat gga 144Asp
Asn Ser Arg Arg Ile Ser Glu Leu Ser Ile Pro Gly Thr His Gly 35
40 45tca atg gca ctt cat ggt gga gtt gct
gga act ata gga gat ata gct 192Ser Met Ala Leu His Gly Gly Val Ala
Gly Thr Ile Gly Asp Ile Ala 50 55
60atc aat caa aca atg aac ctt gaa act caa tta aac tca gga atc cga
240Ile Asn Gln Thr Met Asn Leu Glu Thr Gln Leu Asn Ser Gly Ile Arg65
70 75 80tat att gat att cga
tgt agg cat tat cat aat aat ttt gct atc cat 288Tyr Ile Asp Ile Arg
Cys Arg His Tyr His Asn Asn Phe Ala Ile His 85
90 95cat gga cag ata tac cag cac gca ttt ttt ggt
tct cat gtt tta gag 336His Gly Gln Ile Tyr Gln His Ala Phe Phe Gly
Ser His Val Leu Glu 100 105
110ccc gtg ata aga ttt tta agg caa aat ccc agt gaa aca att tta atg
384Pro Val Ile Arg Phe Leu Arg Gln Asn Pro Ser Glu Thr Ile Leu Met
115 120 125cgt att caa caa gaa tac aat
cca aca ggt aat aca aga act ttt gct 432Arg Ile Gln Gln Glu Tyr Asn
Pro Thr Gly Asn Thr Arg Thr Phe Ala 130 135
140gaa act ttt gaa tcc ttc tgg act cca aat caa cgt tat ttt tgg tct
480Glu Thr Phe Glu Ser Phe Trp Thr Pro Asn Gln Arg Tyr Phe Trp Ser145
150 155 160cct act agt aat
aat cca aca tta gga gac gtc cga gga aga att att 528Pro Thr Ser Asn
Asn Pro Thr Leu Gly Asp Val Arg Gly Arg Ile Ile 165
170 175cta tta caa caa ttc ccc tct gat aga gga
tgg ttt ggt att aat tgg 576Leu Leu Gln Gln Phe Pro Ser Asp Arg Gly
Trp Phe Gly Ile Asn Trp 180 185
190ggc tcg tta gcc ata caa gat cag tgg gag gta gca ggt ctt aat ggc
624Gly Ser Leu Ala Ile Gln Asp Gln Trp Glu Val Ala Gly Leu Asn Gly
195 200 205ata tac aga aaa tgg ata gct
att aaa aat cat ttt ttt aac aca ata 672Ile Tyr Arg Lys Trp Ile Ala
Ile Lys Asn His Phe Phe Asn Thr Ile 210 215
220aat aat aga aat cga atc cat att aat cat tta agt gga act ggt ggt
720Asn Asn Arg Asn Arg Ile His Ile Asn His Leu Ser Gly Thr Gly Gly225
230 235 240ttc ggt gaa cca
aga ccc tgg ttt tta gca agt ggc tat aat agt cga 768Phe Gly Glu Pro
Arg Pro Trp Phe Leu Ala Ser Gly Tyr Asn Ser Arg 245
250 255aat gac aat agt atg tta agg tct gca tcg
aga ggt cct agt gat gga 816Asn Asp Asn Ser Met Leu Arg Ser Ala Ser
Arg Gly Pro Ser Asp Gly 260 265
270tgg cca gat ttc cca cgt cat agg ccc aca gga gat ata tat ttt gga
864Trp Pro Asp Phe Pro Arg His Arg Pro Thr Gly Asp Ile Tyr Phe Gly
275 280 285gga atg aat atc ctt gca act
aga aga att cga gac cgt aga ttt act 912Gly Met Asn Ile Leu Ala Thr
Arg Arg Ile Arg Asp Arg Arg Phe Thr 290 295
300cat aca gga att gtt gct gct gat ttc cct gga aga gga tta att gaa
960His Thr Gly Ile Val Ala Ala Asp Phe Pro Gly Arg Gly Leu Ile Glu305
310 315 320cgc aca att gcg
tta aat ttc cct ata tca cca cct cac ttt cct ggt 1008Arg Thr Ile Ala
Leu Asn Phe Pro Ile Ser Pro Pro His Phe Pro Gly 325
330 335tac tct caa att gtg aca gct tta aat agt
agt agt gta att gat cta 1056Tyr Ser Gln Ile Val Thr Ala Leu Asn Ser
Ser Ser Val Ile Asp Leu 340 345
350aat cct aat cgt aat gtc aca tta tgg tcg aat cat cga gga ctt aat
1104Asn Pro Asn Arg Asn Val Thr Leu Trp Ser Asn His Arg Gly Leu Asn
355 360 365caa agg tgg cgc att caa cat
att tct gga aat tca tat tca ata ttg 1152Gln Arg Trp Arg Ile Gln His
Ile Ser Gly Asn Ser Tyr Ser Ile Leu 370 375
380cca aat ctt cca gtt tta ggt tta cct tat gtt tta act ggg aat gtt
1200Pro Asn Leu Pro Val Leu Gly Leu Pro Tyr Val Leu Thr Gly Asn Val385
390 395 400tat aat ctt aat
tct aat gta ttt ctt gct cga tct aac gga ttg cct 1248Tyr Asn Leu Asn
Ser Asn Val Phe Leu Ala Arg Ser Asn Gly Leu Pro 405
410 415gag caa cag tgg ctt ctg gaa gaa ttt ttt
gac ggt gac tat ata att 1296Glu Gln Gln Trp Leu Leu Glu Glu Phe Phe
Asp Gly Asp Tyr Ile Ile 420 425
430aaa aat aga aga aat ccg aat ctg gtt tta gat gta tcc aga agc agc
1344Lys Asn Arg Arg Asn Pro Asn Leu Val Leu Asp Val Ser Arg Ser Ser
435 440 445act agt aat ggc tca ggt att
att tta agt aca aga cac aat gga aat 1392Thr Ser Asn Gly Ser Gly Ile
Ile Leu Ser Thr Arg His Asn Gly Asn 450 455
460aat caa aga ttt ttt atc aga cca ttt taa
1422Asn Gln Arg Phe Phe Ile Arg Pro Phe 465
4709473PRTBacillus thuringiensis 9Met Val Asp Arg Asn Gly Met Pro Thr Ile
Asp Arg Ser Gly Phe Thr1 5 10
15 Val Asn Arg Thr Val Asn Tyr Thr Asn Thr Asn Trp Met Ala Asn
Ile 20 25 30 Asp
Asn Ser Arg Arg Ile Ser Glu Leu Ser Ile Pro Gly Thr His Gly 35
40 45 Ser Met Ala Leu His Gly
Gly Val Ala Gly Thr Ile Gly Asp Ile Ala 50 55
60 Ile Asn Gln Thr Met Asn Leu Glu Thr Gln Leu
Asn Ser Gly Ile Arg65 70 75
80 Tyr Ile Asp Ile Arg Cys Arg His Tyr His Asn Asn Phe Ala Ile His
85 90 95 His Gly Gln
Ile Tyr Gln His Ala Phe Phe Gly Ser His Val Leu Glu 100
105 110 Pro Val Ile Arg Phe Leu Arg Gln
Asn Pro Ser Glu Thr Ile Leu Met 115 120
125 Arg Ile Gln Gln Glu Tyr Asn Pro Thr Gly Asn Thr Arg
Thr Phe Ala 130 135 140
Glu Thr Phe Glu Ser Phe Trp Thr Pro Asn Gln Arg Tyr Phe Trp Ser145
150 155 160 Pro Thr Ser Asn Asn
Pro Thr Leu Gly Asp Val Arg Gly Arg Ile Ile 165
170 175 Leu Leu Gln Gln Phe Pro Ser Asp Arg Gly
Trp Phe Gly Ile Asn Trp 180 185
190 Gly Ser Leu Ala Ile Gln Asp Gln Trp Glu Val Ala Gly Leu Asn
Gly 195 200 205 Ile
Tyr Arg Lys Trp Ile Ala Ile Lys Asn His Phe Phe Asn Thr Ile 210
215 220 Asn Asn Arg Asn Arg Ile
His Ile Asn His Leu Ser Gly Thr Gly Gly225 230
235 240 Phe Gly Glu Pro Arg Pro Trp Phe Leu Ala Ser
Gly Tyr Asn Ser Arg 245 250
255 Asn Asp Asn Ser Met Leu Arg Ser Ala Ser Arg Gly Pro Ser Asp Gly
260 265 270 Trp Pro Asp
Phe Pro Arg His Arg Pro Thr Gly Asp Ile Tyr Phe Gly 275
280 285 Gly Met Asn Ile Leu Ala Thr Arg
Arg Ile Arg Asp Arg Arg Phe Thr 290 295
300 His Thr Gly Ile Val Ala Ala Asp Phe Pro Gly Arg Gly
Leu Ile Glu305 310 315
320 Arg Thr Ile Ala Leu Asn Phe Pro Ile Ser Pro Pro His Phe Pro Gly
325 330 335 Tyr Ser Gln Ile
Val Thr Ala Leu Asn Ser Ser Ser Val Ile Asp Leu 340
345 350 Asn Pro Asn Arg Asn Val Thr Leu Trp
Ser Asn His Arg Gly Leu Asn 355 360
365 Gln Arg Trp Arg Ile Gln His Ile Ser Gly Asn Ser Tyr Ser
Ile Leu 370 375 380
Pro Asn Leu Pro Val Leu Gly Leu Pro Tyr Val Leu Thr Gly Asn Val385
390 395 400 Tyr Asn Leu Asn Ser
Asn Val Phe Leu Ala Arg Ser Asn Gly Leu Pro 405
410 415 Glu Gln Gln Trp Leu Leu Glu Glu Phe Phe
Asp Gly Asp Tyr Ile Ile 420 425
430 Lys Asn Arg Arg Asn Pro Asn Leu Val Leu Asp Val Ser Arg Ser
Ser 435 440 445 Thr
Ser Asn Gly Ser Gly Ile Ile Leu Ser Thr Arg His Asn Gly Asn 450
455 460 Asn Gln Arg Phe Phe Ile
Arg Pro Phe465 470 101053DNABacillus
thuringiensisCDS(1)...(1053) 10atg aaa tct tat aaa aaa ata tta cta gtt
gcc cca tta gca tgt acc 48Met Lys Ser Tyr Lys Lys Ile Leu Leu Val
Ala Pro Leu Ala Cys Thr1 5 10
15tta gca acg gga gtt ttt aca aca cct cat gca gca ttt gca gca tct
96Leu Ala Thr Gly Val Phe Thr Thr Pro His Ala Ala Phe Ala Ala Ser
20 25 30tca gtg ggc atg cag gcg
aat gca aag gat act ctt gca ttt aat gat 144Ser Val Gly Met Gln Ala
Asn Ala Lys Asp Thr Leu Ala Phe Asn Asp 35 40
45caa caa tta aaa aaa gat tta tca gat cga cta aca tct gcc
gtg aga 192Gln Gln Leu Lys Lys Asp Leu Ser Asp Arg Leu Thr Ser Ala
Val Arg 50 55 60aat agg cca gat tta
ttt ggg ata acc aca cca gtc ctc gtt aat aac 240Asn Arg Pro Asp Leu
Phe Gly Ile Thr Thr Pro Val Leu Val Asn Asn65 70
75 80att aaa aac atg aat ttc aaa tta aca gat
atg aac gcg act tat ggg 288Ile Lys Asn Met Asn Phe Lys Leu Thr Asp
Met Asn Ala Thr Tyr Gly 85 90
95tat aca aat aat gga aca att caa acg gat gcg aag gtg gat cat tat
336Tyr Thr Asn Asn Gly Thr Ile Gln Thr Asp Ala Lys Val Asp His Tyr
100 105 110ggt gat ggg gga caa gtg
gag tta tta agc tat cgg aat gat acc tct 384Gly Asp Gly Gly Gln Val
Glu Leu Leu Ser Tyr Arg Asn Asp Thr Ser 115 120
125gtg aat caa act ttc aat acc cca gaa aaa agc tta aaa aca
tca gaa 432Val Asn Gln Thr Phe Asn Thr Pro Glu Lys Ser Leu Lys Thr
Ser Glu 130 135 140agt ttt acc tat tca
aat cag gaa ggg gta aaa ttg ggg gta gcc tcg 480Ser Phe Thr Tyr Ser
Asn Gln Glu Gly Val Lys Leu Gly Val Ala Ser145 150
155 160gaa act aaa gtc ggt gta gat att ccc ttt
ata ggt ggg gct gac gaa 528Glu Thr Lys Val Gly Val Asp Ile Pro Phe
Ile Gly Gly Ala Asp Glu 165 170
175acg att aag att tcc agc gag ttt tct tat aac cat act agt tca aat
576Thr Ile Lys Ile Ser Ser Glu Phe Ser Tyr Asn His Thr Ser Ser Asn
180 185 190act agc aca aaa gaa gaa
acg act aca ttt aaa tca caa ccg gtt att 624Thr Ser Thr Lys Glu Glu
Thr Thr Thr Phe Lys Ser Gln Pro Val Ile 195 200
205tgt gta gca gga tat aca aca caa ttt tca gga agc gta caa
aat gct 672Cys Val Ala Gly Tyr Thr Thr Gln Phe Ser Gly Ser Val Gln
Asn Ala 210 215 220gtc ttt tct ggg tca
ttt agt ggc act gca gaa gcg tca ggt gat gtg 720Val Phe Ser Gly Ser
Phe Ser Gly Thr Ala Glu Ala Ser Gly Asp Val225 230
235 240aaa ttc caa gaa gtg aac gag ttg ttc cgt
gta gat acg tct tta ggt 768Lys Phe Gln Glu Val Asn Glu Leu Phe Arg
Val Asp Thr Ser Leu Gly 245 250
255gac aat cca aat att aaa gga cac gca tta tac aat gtc ttt aaa tat
816Asp Asn Pro Asn Ile Lys Gly His Ala Leu Tyr Asn Val Phe Lys Tyr
260 265 270tcg ggc atg cct gtt cca
tca tat gta aaa tta gat gat acg aat aaa 864Ser Gly Met Pro Val Pro
Ser Tyr Val Lys Leu Asp Asp Thr Asn Lys 275 280
285aga gct tta att gaa aat gtg aca tcc acg tat agt gga gtt
ggt ggt 912Arg Ala Leu Ile Glu Asn Val Thr Ser Thr Tyr Ser Gly Val
Gly Gly 290 295 300cat tat tca cgc gta
gaa gtt aaa gtg ttc cca aat acg cgt agt aat 960His Tyr Ser Arg Val
Glu Val Lys Val Phe Pro Asn Thr Arg Ser Asn305 310
315 320gaa gat gca atc aca ata cca tat gca aaa
tat atg caa aaa gta aaa 1008Glu Asp Ala Ile Thr Ile Pro Tyr Ala Lys
Tyr Met Gln Lys Val Lys 325 330
335gat ggt aca cta cag aaa gaa tta gaa cag cat tat aaa aaa gct
1053Asp Gly Thr Leu Gln Lys Glu Leu Glu Gln His Tyr Lys Lys Ala
340 345 35011351PRTBacillus
thuringiensis 11Met Lys Ser Tyr Lys Lys Ile Leu Leu Val Ala Pro Leu Ala
Cys Thr1 5 10 15
Leu Ala Thr Gly Val Phe Thr Thr Pro His Ala Ala Phe Ala Ala Ser
20 25 30 Ser Val Gly Met Gln
Ala Asn Ala Lys Asp Thr Leu Ala Phe Asn Asp 35 40
45 Gln Gln Leu Lys Lys Asp Leu Ser Asp Arg
Leu Thr Ser Ala Val Arg 50 55 60
Asn Arg Pro Asp Leu Phe Gly Ile Thr Thr Pro Val Leu Val Asn
Asn65 70 75 80 Ile
Lys Asn Met Asn Phe Lys Leu Thr Asp Met Asn Ala Thr Tyr Gly
85 90 95 Tyr Thr Asn Asn Gly Thr
Ile Gln Thr Asp Ala Lys Val Asp His Tyr 100
105 110 Gly Asp Gly Gly Gln Val Glu Leu Leu Ser
Tyr Arg Asn Asp Thr Ser 115 120
125 Val Asn Gln Thr Phe Asn Thr Pro Glu Lys Ser Leu Lys Thr
Ser Glu 130 135 140
Ser Phe Thr Tyr Ser Asn Gln Glu Gly Val Lys Leu Gly Val Ala Ser145
150 155 160 Glu Thr Lys Val Gly
Val Asp Ile Pro Phe Ile Gly Gly Ala Asp Glu 165
170 175 Thr Ile Lys Ile Ser Ser Glu Phe Ser Tyr
Asn His Thr Ser Ser Asn 180 185
190 Thr Ser Thr Lys Glu Glu Thr Thr Thr Phe Lys Ser Gln Pro Val
Ile 195 200 205 Cys
Val Ala Gly Tyr Thr Thr Gln Phe Ser Gly Ser Val Gln Asn Ala 210
215 220 Val Phe Ser Gly Ser Phe
Ser Gly Thr Ala Glu Ala Ser Gly Asp Val225 230
235 240 Lys Phe Gln Glu Val Asn Glu Leu Phe Arg Val
Asp Thr Ser Leu Gly 245 250
255 Asp Asn Pro Asn Ile Lys Gly His Ala Leu Tyr Asn Val Phe Lys Tyr
260 265 270 Ser Gly Met
Pro Val Pro Ser Tyr Val Lys Leu Asp Asp Thr Asn Lys 275
280 285 Arg Ala Leu Ile Glu Asn Val Thr
Ser Thr Tyr Ser Gly Val Gly Gly 290 295
300 His Tyr Ser Arg Val Glu Val Lys Val Phe Pro Asn Thr
Arg Ser Asn305 310 315
320 Glu Asp Ala Ile Thr Ile Pro Tyr Ala Lys Tyr Met Gln Lys Val Lys
325 330 335 Asp Gly Thr Leu
Gln Lys Glu Leu Glu Gln His Tyr Lys Lys Ala 340
345 350 121062DNAUnknownCDS(1)...(1062)Isolated from
soil sample 12atg tca gga ttt gaa gaa tta ttt tac cca gat aat acc aac cgt
aga 48Met Ser Gly Phe Glu Glu Leu Phe Tyr Pro Asp Asn Thr Asn Arg
Arg1 5 10 15agc aga gta
gaa caa tta atg gct caa tgt aca gat tta act aac caa 96Ser Arg Val
Glu Gln Leu Met Ala Gln Cys Thr Asp Leu Thr Asn Gln 20
25 30att aag aat gat aga aaa gac att gat gac
ctt ttt cga aat aat gat 144Ile Lys Asn Asp Arg Lys Asp Ile Asp Asp
Leu Phe Arg Asn Asn Asp 35 40
45cca att atc aaa gaa aag ctt aaa ggt aag cta ata tta agc att cct
192Pro Ile Ile Lys Glu Lys Leu Lys Gly Lys Leu Ile Leu Ser Ile Pro 50
55 60aat aaa tat ttt gat ata tcc aaa aat
tcc ata gat gat att tta aat 240Asn Lys Tyr Phe Asp Ile Ser Lys Asn
Ser Ile Asp Asp Ile Leu Asn65 70 75
80tat gct tta ggt cca gtt cct gct gct att gta ggg gct gat
aat aga 288Tyr Ala Leu Gly Pro Val Pro Ala Ala Ile Val Gly Ala Asp
Asn Arg 85 90 95aca tta
tat caa aaa ata att aaa gaa ttg gtg aaa att cgt att gag 336Thr Leu
Tyr Gln Lys Ile Ile Lys Glu Leu Val Lys Ile Arg Ile Glu 100
105 110tta aag tat ata gaa tta aag act att
tat gat aaa atg gtt att agt 384Leu Lys Tyr Ile Glu Leu Lys Thr Ile
Tyr Asp Lys Met Val Ile Ser 115 120
125gcc gtt ata aaa gag att act cgg att aaa aaa aca gcc caa aaa cat
432Ala Val Ile Lys Glu Ile Thr Arg Ile Lys Lys Thr Ala Gln Lys His 130
135 140aac tgg act aat gaa ata cta gca
gaa att act cag gaa atc att gaa 480Asn Trp Thr Asn Glu Ile Leu Ala
Glu Ile Thr Gln Glu Ile Ile Glu145 150
155 160aac act att gaa acg ata gaa aca att aat gta gaa
cct tct aga gaa 528Asn Thr Ile Glu Thr Ile Glu Thr Ile Asn Val Glu
Pro Ser Arg Glu 165 170
175tta gct gta gat aaa tta caa gaa att gat gaa aat ctt gca gct tgg
576Leu Ala Val Asp Lys Leu Gln Glu Ile Asp Glu Asn Leu Ala Ala Trp
180 185 190atc aat gaa gat cca tct
aat caa aaa att act atg gaa ttg aat gca 624Ile Asn Glu Asp Pro Ser
Asn Gln Lys Ile Thr Met Glu Leu Asn Ala 195 200
205tta gat gaa gta tat aaa gtt att tcg cct tta aat aat aaa
agc gtg 672Leu Asp Glu Val Tyr Lys Val Ile Ser Pro Leu Asn Asn Lys
Ser Val 210 215 220tta gat ttt tct cgc
tca aat aat aat gcg att tta tgg gat gac cat 720Leu Asp Phe Ser Arg
Ser Asn Asn Asn Ala Ile Leu Trp Asp Asp His225 230
235 240gat ggt gaa aat caa aaa tgg aaa ttt gaa
tac aat gca aag cac aca 768Asp Gly Glu Asn Gln Lys Trp Lys Phe Glu
Tyr Asn Ala Lys His Thr 245 250
255gca tac caa att aaa agt ctg gta aat aaa gac ttt gtt tta gca tgg
816Ala Tyr Gln Ile Lys Ser Leu Val Asn Lys Asp Phe Val Leu Ala Trp
260 265 270gat gac ggt aat aaa tta
aag aat gtg ttt gtt aca aaa aat caa tac 864Asp Asp Gly Asn Lys Leu
Lys Asn Val Phe Val Thr Lys Asn Gln Tyr 275 280
285aaa gaa gaa cat ttt tgg att tta gaa aag aca gaa gat gac
aat tat 912Lys Glu Glu His Phe Trp Ile Leu Glu Lys Thr Glu Asp Asp
Asn Tyr 290 295 300att ata aaa aat aaa
aaa tcc cta att tta ata tta gag gtc gat agg 960Ile Ile Lys Asn Lys
Lys Ser Leu Ile Leu Ile Leu Glu Val Asp Arg305 310
315 320gct caa act aat aat gga gca aac att aaa
tta aat gaa caa aac cgt 1008Ala Gln Thr Asn Asn Gly Ala Asn Ile Lys
Leu Asn Glu Gln Asn Arg 325 330
335ata gat aaa cgg tta att aat gct caa aaa ttc aaa tta gca aag tgt
1056Ile Asp Lys Arg Leu Ile Asn Ala Gln Lys Phe Lys Leu Ala Lys Cys
340 345 350tat taa
1062Tyr
13353PRTUnknownIsolated from soil sample 13Met Ser Gly Phe Glu Glu Leu
Phe Tyr Pro Asp Asn Thr Asn Arg Arg1 5 10
15 Ser Arg Val Glu Gln Leu Met Ala Gln Cys Thr Asp
Leu Thr Asn Gln 20 25 30
Ile Lys Asn Asp Arg Lys Asp Ile Asp Asp Leu Phe Arg Asn Asn Asp
35 40 45 Pro Ile Ile Lys
Glu Lys Leu Lys Gly Lys Leu Ile Leu Ser Ile Pro 50 55
60 Asn Lys Tyr Phe Asp Ile Ser Lys Asn
Ser Ile Asp Asp Ile Leu Asn65 70 75
80 Tyr Ala Leu Gly Pro Val Pro Ala Ala Ile Val Gly Ala Asp
Asn Arg 85 90 95
Thr Leu Tyr Gln Lys Ile Ile Lys Glu Leu Val Lys Ile Arg Ile Glu
100 105 110 Leu Lys Tyr Ile Glu
Leu Lys Thr Ile Tyr Asp Lys Met Val Ile Ser 115
120 125 Ala Val Ile Lys Glu Ile Thr Arg Ile
Lys Lys Thr Ala Gln Lys His 130 135
140 Asn Trp Thr Asn Glu Ile Leu Ala Glu Ile Thr Gln Glu
Ile Ile Glu145 150 155
160 Asn Thr Ile Glu Thr Ile Glu Thr Ile Asn Val Glu Pro Ser Arg Glu
165 170 175 Leu Ala Val Asp
Lys Leu Gln Glu Ile Asp Glu Asn Leu Ala Ala Trp 180
185 190 Ile Asn Glu Asp Pro Ser Asn Gln Lys
Ile Thr Met Glu Leu Asn Ala 195 200
205 Leu Asp Glu Val Tyr Lys Val Ile Ser Pro Leu Asn Asn Lys
Ser Val 210 215 220
Leu Asp Phe Ser Arg Ser Asn Asn Asn Ala Ile Leu Trp Asp Asp His225
230 235 240 Asp Gly Glu Asn Gln
Lys Trp Lys Phe Glu Tyr Asn Ala Lys His Thr 245
250 255 Ala Tyr Gln Ile Lys Ser Leu Val Asn Lys
Asp Phe Val Leu Ala Trp 260 265
270 Asp Asp Gly Asn Lys Leu Lys Asn Val Phe Val Thr Lys Asn Gln
Tyr 275 280 285 Lys
Glu Glu His Phe Trp Ile Leu Glu Lys Thr Glu Asp Asp Asn Tyr 290
295 300 Ile Ile Lys Asn Lys Lys
Ser Leu Ile Leu Ile Leu Glu Val Asp Arg305 310
315 320 Ala Gln Thr Asn Asn Gly Ala Asn Ile Lys Leu
Asn Glu Gln Asn Arg 325 330
335 Ile Asp Lys Arg Leu Ile Asn Ala Gln Lys Phe Lys Leu Ala Lys Cys
340 345 350
Tyr14942DNABacillus thuringiensisCDS(1)...(942) 14atg aaa aaa ata gga ttc
gtt agt ggc atg aca tct tta gct att tta 48Met Lys Lys Ile Gly Phe
Val Ser Gly Met Thr Ser Leu Ala Ile Leu1 5
10 15gct agt gct ttt gga gga gaa atc aaa cct gca tct
gcg agc act tct 96Ala Ser Ala Phe Gly Gly Glu Ile Lys Pro Ala Ser
Ala Ser Thr Ser 20 25 30gct
tta tcc caa caa tct act ctt cat tct aca caa caa gct gtt cag 144Ala
Leu Ser Gln Gln Ser Thr Leu His Ser Thr Gln Gln Ala Val Gln 35
40 45aaa cat aat ttt gat gca atc gtt aat
gag atc att cat gca tgg att 192Lys His Asn Phe Asp Ala Ile Val Asn
Glu Ile Ile His Ala Trp Ile 50 55
60aaa aaa aat aat cct ggt gga cat tat cca ttt att gat agt aaa aat
240Lys Lys Asn Asn Pro Gly Gly His Tyr Pro Phe Ile Asp Ser Lys Asn65
70 75 80atc tca caa att aaa
gaa tcc cgt tta aac att ggt gct cct aaa cga 288Ile Ser Gln Ile Lys
Glu Ser Arg Leu Asn Ile Gly Ala Pro Lys Arg 85
90 95att gga aat att gaa aat cga aca tta gtt aca
act att cct tct aga 336Ile Gly Asn Ile Glu Asn Arg Thr Leu Val Thr
Thr Ile Pro Ser Arg 100 105
110att tac aat aat aca aat gca tct gta tta aaa cca gca act tca acc
384Ile Tyr Asn Asn Thr Asn Ala Ser Val Leu Lys Pro Ala Thr Ser Thr
115 120 125gaa tta aaa aca agt cac agt
agc tct ttt act aat tta aca gaa atc 432Glu Leu Lys Thr Ser His Ser
Ser Ser Phe Thr Asn Leu Thr Glu Ile 130 135
140aca cat aca ggt ggc atc aca aca aaa gcg gaa gtg aaa ttt aaa cca
480Thr His Thr Gly Gly Ile Thr Thr Lys Ala Glu Val Lys Phe Lys Pro145
150 155 160aaa ggc ctt gtt
gca gat ggt gaa gtt tct aca ggt cta gaa tta aaa 528Lys Gly Leu Val
Ala Asp Gly Glu Val Ser Thr Gly Leu Glu Leu Lys 165
170 175tat gaa tat agt aat aca cag gga acc aat
caa acc caa aca aca acc 576Tyr Glu Tyr Ser Asn Thr Gln Gly Thr Asn
Gln Thr Gln Thr Thr Thr 180 185
190aat gaa tta agt ttt aaa gtc gat aca cct gtt gaa gtt cct cca cgg
624Asn Glu Leu Ser Phe Lys Val Asp Thr Pro Val Glu Val Pro Pro Arg
195 200 205tca tct ata gaa gta ata aca
aat att tat aaa gac aaa gta aga tat 672Ser Ser Ile Glu Val Ile Thr
Asn Ile Tyr Lys Asp Lys Val Arg Tyr 210 215
220gaa tat aca gga tat agt gaa ttc aca ggt gaa gtc aca ttc caa tat
720Glu Tyr Thr Gly Tyr Ser Glu Phe Thr Gly Glu Val Thr Phe Gln Tyr225
230 235 240aga ttg aat gca
tct gac aca cca aaa aca gta aca aga gaa att ggt 768Arg Leu Asn Ala
Ser Asp Thr Pro Lys Thr Val Thr Arg Glu Ile Gly 245
250 255acc atg atg tat gaa ata gat gat gaa acg
tac aat aaa tta gct gac 816Thr Met Met Tyr Glu Ile Asp Asp Glu Thr
Tyr Asn Lys Leu Ala Asp 260 265
270aga gga att aca gta aaa ggc gca gta gat tct cca gat gtc tta cgt
864Arg Gly Ile Thr Val Lys Gly Ala Val Asp Ser Pro Asp Val Leu Arg
275 280 285att aaa gga aca gct att ttg
gat gta gat gag gca tat agc acg gaa 912Ile Lys Gly Thr Ala Ile Leu
Asp Val Asp Glu Ala Tyr Ser Thr Glu 290 295
300gta att gca aga gat att gca cct att cag
942Val Ile Ala Arg Asp Ile Ala Pro Ile Gln305
31015314PRTBacillus thuringiensis 15Met Lys Lys Ile Gly Phe Val Ser Gly
Met Thr Ser Leu Ala Ile Leu1 5 10
15 Ala Ser Ala Phe Gly Gly Glu Ile Lys Pro Ala Ser Ala Ser
Thr Ser 20 25 30
Ala Leu Ser Gln Gln Ser Thr Leu His Ser Thr Gln Gln Ala Val Gln 35
40 45 Lys His Asn Phe Asp
Ala Ile Val Asn Glu Ile Ile His Ala Trp Ile 50 55
60 Lys Lys Asn Asn Pro Gly Gly His Tyr Pro
Phe Ile Asp Ser Lys Asn65 70 75
80 Ile Ser Gln Ile Lys Glu Ser Arg Leu Asn Ile Gly Ala Pro Lys
Arg 85 90 95 Ile
Gly Asn Ile Glu Asn Arg Thr Leu Val Thr Thr Ile Pro Ser Arg
100 105 110 Ile Tyr Asn Asn Thr
Asn Ala Ser Val Leu Lys Pro Ala Thr Ser Thr 115
120 125 Glu Leu Lys Thr Ser His Ser Ser Ser
Phe Thr Asn Leu Thr Glu Ile 130 135
140 Thr His Thr Gly Gly Ile Thr Thr Lys Ala Glu Val Lys
Phe Lys Pro145 150 155
160 Lys Gly Leu Val Ala Asp Gly Glu Val Ser Thr Gly Leu Glu Leu Lys
165 170 175 Tyr Glu Tyr Ser
Asn Thr Gln Gly Thr Asn Gln Thr Gln Thr Thr Thr 180
185 190 Asn Glu Leu Ser Phe Lys Val Asp Thr
Pro Val Glu Val Pro Pro Arg 195 200
205 Ser Ser Ile Glu Val Ile Thr Asn Ile Tyr Lys Asp Lys Val
Arg Tyr 210 215 220
Glu Tyr Thr Gly Tyr Ser Glu Phe Thr Gly Glu Val Thr Phe Gln Tyr225
230 235 240 Arg Leu Asn Ala Ser
Asp Thr Pro Lys Thr Val Thr Arg Glu Ile Gly 245
250 255 Thr Met Met Tyr Glu Ile Asp Asp Glu Thr
Tyr Asn Lys Leu Ala Asp 260 265
270 Arg Gly Ile Thr Val Lys Gly Ala Val Asp Ser Pro Asp Val Leu
Arg 275 280 285 Ile
Lys Gly Thr Ala Ile Leu Asp Val Asp Glu Ala Tyr Ser Thr Glu 290
295 300 Val Ile Ala Arg Asp Ile
Ala Pro Ile Gln305 310 163552DNABacillus
thuringiensisCDS(1)...(3552) 16atg tgt ttt cta tgt cat aga aga cac ata
gga gat tgt tta ttc aat 48Met Cys Phe Leu Cys His Arg Arg His Ile
Gly Asp Cys Leu Phe Asn1 5 10
15tta ata tac ttt ttg gag gta aat tat atg gat ata aga aat cag aag
96Leu Ile Tyr Phe Leu Glu Val Asn Tyr Met Asp Ile Arg Asn Gln Lys
20 25 30aaa tca gaa gaa ata cac
ccc aca tta aat gaa tca tct tct aat aca 144Lys Ser Glu Glu Ile His
Pro Thr Leu Asn Glu Ser Ser Ser Asn Thr 35 40
45aca tca aag tat cca ctt gca agt gat cta atc aaa caa tat
caa aat 192Thr Ser Lys Tyr Pro Leu Ala Ser Asp Leu Ile Lys Gln Tyr
Gln Asn 50 55 60atg aat tat aaa gat
agt tta aat ata atc gag gag aat aat gta atc 240Met Asn Tyr Lys Asp
Ser Leu Asn Ile Ile Glu Glu Asn Asn Val Ile65 70
75 80gat cct gta tct gga act gcc gct ttg gta
act gca agg aaa att ggt 288Asp Pro Val Ser Gly Thr Ala Ala Leu Val
Thr Ala Arg Lys Ile Gly 85 90
95ggt aag ata att aag aag tta gga gag gca gct ctg tct aaa att ttg
336Gly Lys Ile Ile Lys Lys Leu Gly Glu Ala Ala Leu Ser Lys Ile Leu
100 105 110aaa gag atc ttg ggt tat
tta tgg ccg act tca gga gaa gac gat tca 384Lys Glu Ile Leu Gly Tyr
Leu Trp Pro Thr Ser Gly Glu Asp Asp Ser 115 120
125tgg aca gat atg atg acg gca gta gaa gaa ctt att gat caa
aaa ata 432Trp Thr Asp Met Met Thr Ala Val Glu Glu Leu Ile Asp Gln
Lys Ile 130 135 140cag gaa cag gta gaa
ttg gat gca ctt acg gag ttg gaa aaa ata ggt 480Gln Glu Gln Val Glu
Leu Asp Ala Leu Thr Glu Leu Glu Lys Ile Gly145 150
155 160gaa gcc gtg gag gca tat tat atc gca tta
gac gat tgg gaa cca gta 528Glu Ala Val Glu Ala Tyr Tyr Ile Ala Leu
Asp Asp Trp Glu Pro Val 165 170
175cct gaa gat atg ttt agt cta agc gaa gta ata aaa cga tac gat ttt
576Pro Glu Asp Met Phe Ser Leu Ser Glu Val Ile Lys Arg Tyr Asp Phe
180 185 190gcg cag caa ctt gca aga
gct tca atg cca gaa ttc aag aag aaa ggt 624Ala Gln Gln Leu Ala Arg
Ala Ser Met Pro Glu Phe Lys Lys Lys Gly 195 200
205tat gaa att ccc ttg tta gca aca tat gca aat gct gca aat
gtt cat 672Tyr Glu Ile Pro Leu Leu Ala Thr Tyr Ala Asn Ala Ala Asn
Val His 210 215 220ttg ctt tta atg aga
gat atg caa ata tac ggg gaa aga tgg gga ata 720Leu Leu Leu Met Arg
Asp Met Gln Ile Tyr Gly Glu Arg Trp Gly Ile225 230
235 240cct aaa gaa gat ata gag ctc tac tta tct
gaa caa gaa aat ttt acc 768Pro Lys Glu Asp Ile Glu Leu Tyr Leu Ser
Glu Gln Glu Asn Phe Thr 245 250
255tct gaa tat aca gat cat tgc gta aaa tat tat aat gag gga tta aat
816Ser Glu Tyr Thr Asp His Cys Val Lys Tyr Tyr Asn Glu Gly Leu Asn
260 265 270caa ttg aaa aat aaa agt
ggc gta agt ggt tta gtt tgg gag aat tat 864Gln Leu Lys Asn Lys Ser
Gly Val Ser Gly Leu Val Trp Glu Asn Tyr 275 280
285aac cgt ttc cgt aca gaa atg aca atc ctg gtg tta gat att
gtg gct 912Asn Arg Phe Arg Thr Glu Met Thr Ile Leu Val Leu Asp Ile
Val Ala 290 295 300gta ttt cca cgc tac
aat gta atc gaa tat cct ata gat tca aca gta 960Val Phe Pro Arg Tyr
Asn Val Ile Glu Tyr Pro Ile Asp Ser Thr Val305 310
315 320gaa tta aca aga aca att tat cta gat cca
ctt ggt tac aca ggg aat 1008Glu Leu Thr Arg Thr Ile Tyr Leu Asp Pro
Leu Gly Tyr Thr Gly Asn 325 330
335tcc aat gac gag cat ccc gaa tat tat gcg tct aca aaa tca ttt tca
1056Ser Asn Asp Glu His Pro Glu Tyr Tyr Ala Ser Thr Lys Ser Phe Ser
340 345 350tca ata gag agt aga gcc
ata cct gca ccc aca tta ttc cag tgg atc 1104Ser Ile Glu Ser Arg Ala
Ile Pro Ala Pro Thr Leu Phe Gln Trp Ile 355 360
365act gaa ctt caa gta tat tca gca aaa ggc tct cat ggt tct
acc tat 1152Thr Glu Leu Gln Val Tyr Ser Ala Lys Gly Ser His Gly Ser
Thr Tyr 370 375 380act aca tgg tgg act
gga cat aaa gtg act gct aag cct act aat ggt 1200Thr Thr Trp Trp Thr
Gly His Lys Val Thr Ala Lys Pro Thr Asn Gly385 390
395 400ggt ctt gaa agt aaa tat gat ttc gga agt
tct tca ggt tct cag aac 1248Gly Leu Glu Ser Lys Tyr Asp Phe Gly Ser
Ser Ser Gly Ser Gln Asn 405 410
415aag gat gtt ttt gct ctt gat ggc aag gat gta tat gat tca caa agt
1296Lys Asp Val Phe Ala Leu Asp Gly Lys Asp Val Tyr Asp Ser Gln Ser
420 425 430atg tta aca tcg att agt
tat tcc ggt att aga tat ttt ggg tgt cct 1344Met Leu Thr Ser Ile Ser
Tyr Ser Gly Ile Arg Tyr Phe Gly Cys Pro 435 440
445cag ttt aag tta aat tgg ata aat aag aat aat gag cta gca
gaa cag 1392Gln Phe Lys Leu Asn Trp Ile Asn Lys Asn Asn Glu Leu Ala
Glu Gln 450 455 460ata ttt aat tat tca
agt aat gtt ggt tca tct ttc agt gag tat agg 1440Ile Phe Asn Tyr Ser
Ser Asn Val Gly Ser Ser Phe Ser Glu Tyr Arg465 470
475 480tat agc aag gat gaa tta cca ata gaa ttg
ttg gcg agc cct att tat 1488Tyr Ser Lys Asp Glu Leu Pro Ile Glu Leu
Leu Ala Ser Pro Ile Tyr 485 490
495gga gat att gag gaa tac agt cat agg tta agt cac gtt tca gag gta
1536Gly Asp Ile Glu Glu Tyr Ser His Arg Leu Ser His Val Ser Glu Val
500 505 510att aaa gat tat ggg cag
gga ata att cct gta tta ggt ttc aca cat 1584Ile Lys Asp Tyr Gly Gln
Gly Ile Ile Pro Val Leu Gly Phe Thr His 515 520
525gta agt gta agt cgt gac aat aga att tat tca gat aag att
acg caa 1632Val Ser Val Ser Arg Asp Asn Arg Ile Tyr Ser Asp Lys Ile
Thr Gln 530 535 540att cca gct gtg aaa
atg tat gaa tta gta agc cca gcc gtt gtt gta 1680Ile Pro Ala Val Lys
Met Tyr Glu Leu Val Ser Pro Ala Val Val Val545 550
555 560aaa ggg cct gga tct aca ggt gga gat tta
gtt aag aga ggg agt agt 1728Lys Gly Pro Gly Ser Thr Gly Gly Asp Leu
Val Lys Arg Gly Ser Ser 565 570
575ggt aat ata gga tct atg aat gtt acc gta aac tca cca ctt tct caa
1776Gly Asn Ile Gly Ser Met Asn Val Thr Val Asn Ser Pro Leu Ser Gln
580 585 590aaa tat cgt gtc aga gtt
cga tat gcc act aat gct tct ggc cag tta 1824Lys Tyr Arg Val Arg Val
Arg Tyr Ala Thr Asn Ala Ser Gly Gln Leu 595 600
605aac gtg agt att aac gat aaa tta aca ctt caa aaa cct ttt
gaa aga 1872Asn Val Ser Ile Asn Asp Lys Leu Thr Leu Gln Lys Pro Phe
Glu Arg 610 615 620acc gga aca aca ata
ggt gaa gga aca gat ttg tcc tat gat tca ttt 1920Thr Gly Thr Thr Ile
Gly Glu Gly Thr Asp Leu Ser Tyr Asp Ser Phe625 630
635 640gga tat tta gaa tat tct acg acg att caa
ttt ccg aat gag cac cca 1968Gly Tyr Leu Glu Tyr Ser Thr Thr Ile Gln
Phe Pro Asn Glu His Pro 645 650
655aaa atc act ttt aat tta tcc cat tgg agc ggc agt gga gca ttt tat
2016Lys Ile Thr Phe Asn Leu Ser His Trp Ser Gly Ser Gly Ala Phe Tyr
660 665 670ata gat aaa atc gaa ttt
atc cct gta gat gaa aat tac gat gaa aga 2064Ile Asp Lys Ile Glu Phe
Ile Pro Val Asp Glu Asn Tyr Asp Glu Arg 675 680
685gta aca cta gaa aaa gca cag aaa gcc gtg aat gcc ttg ttt
aca gcg 2112Val Thr Leu Glu Lys Ala Gln Lys Ala Val Asn Ala Leu Phe
Thr Ala 690 695 700gga aga aat gca ctc
caa aaa gat gtg aca gat ttt aaa gta gat cag 2160Gly Arg Asn Ala Leu
Gln Lys Asp Val Thr Asp Phe Lys Val Asp Gln705 710
715 720gtt tcc att tta gtg gat tgt ata cca ggg
gaa ttg tat cca aat gag 2208Val Ser Ile Leu Val Asp Cys Ile Pro Gly
Glu Leu Tyr Pro Asn Glu 725 730
735aaa cgc gaa cta cta agt tta gtc aaa tac gca aaa cgg ttg agt tat
2256Lys Arg Glu Leu Leu Ser Leu Val Lys Tyr Ala Lys Arg Leu Ser Tyr
740 745 750tcc cgt aat tta ctc cta
gac cca aca ttc gat tct atc aat tca cca 2304Ser Arg Asn Leu Leu Leu
Asp Pro Thr Phe Asp Ser Ile Asn Ser Pro 755 760
765gat gag aat ggc tgg tac ggg agt aat ggt att gca att gga
aat ggg 2352Asp Glu Asn Gly Trp Tyr Gly Ser Asn Gly Ile Ala Ile Gly
Asn Gly 770 775 780aac ttt gta ttc aaa
gga aac tat tta att ttc tca ggt acc aat gat 2400Asn Phe Val Phe Lys
Gly Asn Tyr Leu Ile Phe Ser Gly Thr Asn Asp785 790
795 800aca caa tac cca acg tat ctc tat caa aaa
att gat gaa tcc aag ctc 2448Thr Gln Tyr Pro Thr Tyr Leu Tyr Gln Lys
Ile Asp Glu Ser Lys Leu 805 810
815aaa gaa tat aca cgc tat aaa ctg aga ggt ttt atc gag agt agt cag
2496Lys Glu Tyr Thr Arg Tyr Lys Leu Arg Gly Phe Ile Glu Ser Ser Gln
820 825 830gat tta gag gca tat gtg
att cgc tat gat gca aaa cat gaa aca ttg 2544Asp Leu Glu Ala Tyr Val
Ile Arg Tyr Asp Ala Lys His Glu Thr Leu 835 840
845gat gta tcc aat aat cta ttc cca gat att tct cct gtg aat
gca tgc 2592Asp Val Ser Asn Asn Leu Phe Pro Asp Ile Ser Pro Val Asn
Ala Cys 850 855 860gga gaa cca aat cgt
tgt gcg gca cta caa tac ctg gat gaa aac cca 2640Gly Glu Pro Asn Arg
Cys Ala Ala Leu Gln Tyr Leu Asp Glu Asn Pro865 870
875 880agg tta gaa tgt agt tcg ata caa gat ggc
att tta tct gat tcg cat 2688Arg Leu Glu Cys Ser Ser Ile Gln Asp Gly
Ile Leu Ser Asp Ser His 885 890
895tca ttt tct ctc aat ata gat aca ggt tct att gat tac gat gag aac
2736Ser Phe Ser Leu Asn Ile Asp Thr Gly Ser Ile Asp Tyr Asp Glu Asn
900 905 910gta ggt att tgg gtg ttg
ttt aaa att tcc aca ccg gaa ggg tat gcg 2784Val Gly Ile Trp Val Leu
Phe Lys Ile Ser Thr Pro Glu Gly Tyr Ala 915 920
925aaa ttt gga aac cta gaa gtg att gaa gat ggc cta gtc att
gga gaa 2832Lys Phe Gly Asn Leu Glu Val Ile Glu Asp Gly Leu Val Ile
Gly Glu 930 935 940gca tta gcc cgt gtg
aaa cgt caa gaa acg aag tgg aga aac aag ttg 2880Ala Leu Ala Arg Val
Lys Arg Gln Glu Thr Lys Trp Arg Asn Lys Leu945 950
955 960aca caa ctg cga acg gaa aca caa gcg att
tat aca cga gca aaa caa 2928Thr Gln Leu Arg Thr Glu Thr Gln Ala Ile
Tyr Thr Arg Ala Lys Gln 965 970
975gcc att gat aat tta ttc aca aat gca cag gac tct cac tta aaa ata
2976Ala Ile Asp Asn Leu Phe Thr Asn Ala Gln Asp Ser His Leu Lys Ile
980 985 990ggt gct aca ttc gcg tca
att gtg gcc gcg cga aag att gtc caa tca 3024Gly Ala Thr Phe Ala Ser
Ile Val Ala Ala Arg Lys Ile Val Gln Ser 995 1000
1005ata cgt gaa gcg tat atg tca tgg tta tct atc gtc cca agt
gta aat 3072Ile Arg Glu Ala Tyr Met Ser Trp Leu Ser Ile Val Pro Ser
Val Asn 1010 1015 1020tat cct att ttc
aca gag ttg aat gaa aga gta cag cga gca ttt cga 3120Tyr Pro Ile Phe
Thr Glu Leu Asn Glu Arg Val Gln Arg Ala Phe Arg1025 1030
1035 1040tta tat gat gta cga aat gtc gta cgt
aat ggc cga ttc ttg agt gga 3168Leu Tyr Asp Val Arg Asn Val Val Arg
Asn Gly Arg Phe Leu Ser Gly 1045 1050
1055gta tcg gat tgg att gtg aca tct gat gta aag gta caa gaa gaa
aat 3216Val Ser Asp Trp Ile Val Thr Ser Asp Val Lys Val Gln Glu Glu
Asn 1060 1065 1070ggg aac aac
gta tta gtt ctt tcc aat tgg gat gca caa gta tta caa 3264Gly Asn Asn
Val Leu Val Leu Ser Asn Trp Asp Ala Gln Val Leu Gln 1075
1080 1085tgt ctg aat ctc tac gaa gac cat ggg tat atc
tta cgc gta aca gca 3312Cys Leu Asn Leu Tyr Glu Asp His Gly Tyr Ile
Leu Arg Val Thr Ala 1090 1095 1100cgt
aag gaa gga ctc gga gaa gga tat ata aca atc act gat gaa gaa 3360Arg
Lys Glu Gly Leu Gly Glu Gly Tyr Ile Thr Ile Thr Asp Glu Glu1105
1110 1115 1120ggg cat aca gat caa ttg
aca ttt ggt gga tgt gag gag ata gat tca 3408Gly His Thr Asp Gln Leu
Thr Phe Gly Gly Cys Glu Glu Ile Asp Ser 1125
1130 1135tcc aat tct ttc gta tct aca ggt tat att aca aaa
gag cta gaa ttc 3456Ser Asn Ser Phe Val Ser Thr Gly Tyr Ile Thr Lys
Glu Leu Glu Phe 1140 1145
1150ttc cca gat aca gag aaa gtg cgc atc gaa att gga gaa aca gaa gga
3504Phe Pro Asp Thr Glu Lys Val Arg Ile Glu Ile Gly Glu Thr Glu Gly
1155 1160 1165aca ttc cag gta gaa agt gtg
gaa tta ttt ttg atg gaa gac ata tgc 3552Thr Phe Gln Val Glu Ser Val
Glu Leu Phe Leu Met Glu Asp Ile Cys 1170 1175
1180171184PRTBacillus thuringiensis 17Met Cys Phe Leu Cys His Arg
Arg His Ile Gly Asp Cys Leu Phe Asn1 5 10
15 Leu Ile Tyr Phe Leu Glu Val Asn Tyr Met Asp Ile
Arg Asn Gln Lys 20 25 30
Lys Ser Glu Glu Ile His Pro Thr Leu Asn Glu Ser Ser Ser Asn Thr
35 40 45 Thr Ser Lys Tyr
Pro Leu Ala Ser Asp Leu Ile Lys Gln Tyr Gln Asn 50 55
60 Met Asn Tyr Lys Asp Ser Leu Asn Ile
Ile Glu Glu Asn Asn Val Ile65 70 75
80 Asp Pro Val Ser Gly Thr Ala Ala Leu Val Thr Ala Arg Lys
Ile Gly 85 90 95
Gly Lys Ile Ile Lys Lys Leu Gly Glu Ala Ala Leu Ser Lys Ile Leu
100 105 110 Lys Glu Ile Leu Gly
Tyr Leu Trp Pro Thr Ser Gly Glu Asp Asp Ser 115
120 125 Trp Thr Asp Met Met Thr Ala Val Glu
Glu Leu Ile Asp Gln Lys Ile 130 135
140 Gln Glu Gln Val Glu Leu Asp Ala Leu Thr Glu Leu Glu
Lys Ile Gly145 150 155
160 Glu Ala Val Glu Ala Tyr Tyr Ile Ala Leu Asp Asp Trp Glu Pro Val
165 170 175 Pro Glu Asp Met
Phe Ser Leu Ser Glu Val Ile Lys Arg Tyr Asp Phe 180
185 190 Ala Gln Gln Leu Ala Arg Ala Ser Met
Pro Glu Phe Lys Lys Lys Gly 195 200
205 Tyr Glu Ile Pro Leu Leu Ala Thr Tyr Ala Asn Ala Ala Asn
Val His 210 215 220
Leu Leu Leu Met Arg Asp Met Gln Ile Tyr Gly Glu Arg Trp Gly Ile225
230 235 240 Pro Lys Glu Asp Ile
Glu Leu Tyr Leu Ser Glu Gln Glu Asn Phe Thr 245
250 255 Ser Glu Tyr Thr Asp His Cys Val Lys Tyr
Tyr Asn Glu Gly Leu Asn 260 265
270 Gln Leu Lys Asn Lys Ser Gly Val Ser Gly Leu Val Trp Glu Asn
Tyr 275 280 285 Asn
Arg Phe Arg Thr Glu Met Thr Ile Leu Val Leu Asp Ile Val Ala 290
295 300 Val Phe Pro Arg Tyr Asn
Val Ile Glu Tyr Pro Ile Asp Ser Thr Val305 310
315 320 Glu Leu Thr Arg Thr Ile Tyr Leu Asp Pro Leu
Gly Tyr Thr Gly Asn 325 330
335 Ser Asn Asp Glu His Pro Glu Tyr Tyr Ala Ser Thr Lys Ser Phe Ser
340 345 350 Ser Ile Glu
Ser Arg Ala Ile Pro Ala Pro Thr Leu Phe Gln Trp Ile 355
360 365 Thr Glu Leu Gln Val Tyr Ser Ala
Lys Gly Ser His Gly Ser Thr Tyr 370 375
380 Thr Thr Trp Trp Thr Gly His Lys Val Thr Ala Lys Pro
Thr Asn Gly385 390 395
400 Gly Leu Glu Ser Lys Tyr Asp Phe Gly Ser Ser Ser Gly Ser Gln Asn
405 410 415 Lys Asp Val Phe
Ala Leu Asp Gly Lys Asp Val Tyr Asp Ser Gln Ser 420
425 430 Met Leu Thr Ser Ile Ser Tyr Ser Gly
Ile Arg Tyr Phe Gly Cys Pro 435 440
445 Gln Phe Lys Leu Asn Trp Ile Asn Lys Asn Asn Glu Leu Ala
Glu Gln 450 455 460
Ile Phe Asn Tyr Ser Ser Asn Val Gly Ser Ser Phe Ser Glu Tyr Arg465
470 475 480 Tyr Ser Lys Asp Glu
Leu Pro Ile Glu Leu Leu Ala Ser Pro Ile Tyr 485
490 495 Gly Asp Ile Glu Glu Tyr Ser His Arg Leu
Ser His Val Ser Glu Val 500 505
510 Ile Lys Asp Tyr Gly Gln Gly Ile Ile Pro Val Leu Gly Phe Thr
His 515 520 525 Val
Ser Val Ser Arg Asp Asn Arg Ile Tyr Ser Asp Lys Ile Thr Gln 530
535 540 Ile Pro Ala Val Lys Met
Tyr Glu Leu Val Ser Pro Ala Val Val Val545 550
555 560 Lys Gly Pro Gly Ser Thr Gly Gly Asp Leu Val
Lys Arg Gly Ser Ser 565 570
575 Gly Asn Ile Gly Ser Met Asn Val Thr Val Asn Ser Pro Leu Ser Gln
580 585 590 Lys Tyr Arg
Val Arg Val Arg Tyr Ala Thr Asn Ala Ser Gly Gln Leu 595
600 605 Asn Val Ser Ile Asn Asp Lys Leu
Thr Leu Gln Lys Pro Phe Glu Arg 610 615
620 Thr Gly Thr Thr Ile Gly Glu Gly Thr Asp Leu Ser Tyr
Asp Ser Phe625 630 635
640 Gly Tyr Leu Glu Tyr Ser Thr Thr Ile Gln Phe Pro Asn Glu His Pro
645 650 655 Lys Ile Thr Phe
Asn Leu Ser His Trp Ser Gly Ser Gly Ala Phe Tyr 660
665 670 Ile Asp Lys Ile Glu Phe Ile Pro Val
Asp Glu Asn Tyr Asp Glu Arg 675 680
685 Val Thr Leu Glu Lys Ala Gln Lys Ala Val Asn Ala Leu Phe
Thr Ala 690 695 700
Gly Arg Asn Ala Leu Gln Lys Asp Val Thr Asp Phe Lys Val Asp Gln705
710 715 720 Val Ser Ile Leu Val
Asp Cys Ile Pro Gly Glu Leu Tyr Pro Asn Glu 725
730 735 Lys Arg Glu Leu Leu Ser Leu Val Lys Tyr
Ala Lys Arg Leu Ser Tyr 740 745
750 Ser Arg Asn Leu Leu Leu Asp Pro Thr Phe Asp Ser Ile Asn Ser
Pro 755 760 765 Asp
Glu Asn Gly Trp Tyr Gly Ser Asn Gly Ile Ala Ile Gly Asn Gly 770
775 780 Asn Phe Val Phe Lys Gly
Asn Tyr Leu Ile Phe Ser Gly Thr Asn Asp785 790
795 800 Thr Gln Tyr Pro Thr Tyr Leu Tyr Gln Lys Ile
Asp Glu Ser Lys Leu 805 810
815 Lys Glu Tyr Thr Arg Tyr Lys Leu Arg Gly Phe Ile Glu Ser Ser Gln
820 825 830 Asp Leu Glu
Ala Tyr Val Ile Arg Tyr Asp Ala Lys His Glu Thr Leu 835
840 845 Asp Val Ser Asn Asn Leu Phe Pro
Asp Ile Ser Pro Val Asn Ala Cys 850 855
860 Gly Glu Pro Asn Arg Cys Ala Ala Leu Gln Tyr Leu Asp
Glu Asn Pro865 870 875
880 Arg Leu Glu Cys Ser Ser Ile Gln Asp Gly Ile Leu Ser Asp Ser His
885 890 895 Ser Phe Ser Leu
Asn Ile Asp Thr Gly Ser Ile Asp Tyr Asp Glu Asn 900
905 910 Val Gly Ile Trp Val Leu Phe Lys Ile
Ser Thr Pro Glu Gly Tyr Ala 915 920
925 Lys Phe Gly Asn Leu Glu Val Ile Glu Asp Gly Leu Val Ile
Gly Glu 930 935 940
Ala Leu Ala Arg Val Lys Arg Gln Glu Thr Lys Trp Arg Asn Lys Leu945
950 955 960 Thr Gln Leu Arg Thr
Glu Thr Gln Ala Ile Tyr Thr Arg Ala Lys Gln 965
970 975 Ala Ile Asp Asn Leu Phe Thr Asn Ala Gln
Asp Ser His Leu Lys Ile 980 985
990 Gly Ala Thr Phe Ala Ser Ile Val Ala Ala Arg Lys Ile Val Gln
Ser 995 1000 1005 Ile
Arg Glu Ala Tyr Met Ser Trp Leu Ser Ile Val Pro Ser Val Asn 1010
1015 1020 Tyr Pro Ile Phe Thr Glu
Leu Asn Glu Arg Val Gln Arg Ala Phe Arg1025 1030
1035 1040 Leu Tyr Asp Val Arg Asn Val Val Arg Asn Gly
Arg Phe Leu Ser Gly 1045 1050
1055 Val Ser Asp Trp Ile Val Thr Ser Asp Val Lys Val Gln Glu Glu Asn
1060 1065 1070 Gly Asn Asn
Val Leu Val Leu Ser Asn Trp Asp Ala Gln Val Leu Gln 1075
1080 1085 Cys Leu Asn Leu Tyr Glu Asp His
Gly Tyr Ile Leu Arg Val Thr Ala 1090 1095
1100 Arg Lys Glu Gly Leu Gly Glu Gly Tyr Ile Thr Ile Thr
Asp Glu Glu1105 1110 1115
1120 Gly His Thr Asp Gln Leu Thr Phe Gly Gly Cys Glu Glu Ile Asp Ser
1125 1130 1135 Ser Asn Ser Phe
Val Ser Thr Gly Tyr Ile Thr Lys Glu Leu Glu Phe 1140
1145 1150 Phe Pro Asp Thr Glu Lys Val Arg Ile
Glu Ile Gly Glu Thr Glu Gly 1155 1160
1165 Thr Phe Gln Val Glu Ser Val Glu Leu Phe Leu Met Glu Asp
Ile Cys 1170 1175 1180
183477DNABacillus thuringiensisCDS(1)...(3477) 18atg gat ata aga aat cag
aag aaa tca gaa gaa ata cac ccc aca tta 48Met Asp Ile Arg Asn Gln
Lys Lys Ser Glu Glu Ile His Pro Thr Leu1 5
10 15aat gaa tca tct tct aat aca aca tca aag tat cca
ctt gca agt gat 96Asn Glu Ser Ser Ser Asn Thr Thr Ser Lys Tyr Pro
Leu Ala Ser Asp 20 25 30cta
atc aaa caa tat caa aat atg aat tat aaa gat agt tta aat ata 144Leu
Ile Lys Gln Tyr Gln Asn Met Asn Tyr Lys Asp Ser Leu Asn Ile 35
40 45atc gag gag aat aat gta atc gat cct
gta tct gga act gcc gct ttg 192Ile Glu Glu Asn Asn Val Ile Asp Pro
Val Ser Gly Thr Ala Ala Leu 50 55
60gta act gca agg aaa att ggt ggt aag ata att aag aag tta gga gag
240Val Thr Ala Arg Lys Ile Gly Gly Lys Ile Ile Lys Lys Leu Gly Glu65
70 75 80gca gct ctg tct aaa
att ttg aaa gag atc ttg ggt tat tta tgg ccg 288Ala Ala Leu Ser Lys
Ile Leu Lys Glu Ile Leu Gly Tyr Leu Trp Pro 85
90 95act tca gga gaa gac gat tca tgg aca gat atg
atg acg gca gta gaa 336Thr Ser Gly Glu Asp Asp Ser Trp Thr Asp Met
Met Thr Ala Val Glu 100 105
110gaa ctt att gat caa aaa ata cag gaa cag gta gaa ttg gat gca ctt
384Glu Leu Ile Asp Gln Lys Ile Gln Glu Gln Val Glu Leu Asp Ala Leu
115 120 125acg gag ttg gaa aaa ata ggt
gaa gcc gtg gag gca tat tat atc gca 432Thr Glu Leu Glu Lys Ile Gly
Glu Ala Val Glu Ala Tyr Tyr Ile Ala 130 135
140tta gac gat tgg gaa cca gta cct gaa gat atg ttt agt cta agc gaa
480Leu Asp Asp Trp Glu Pro Val Pro Glu Asp Met Phe Ser Leu Ser Glu145
150 155 160gta ata aaa cga
tac gat ttt gcg cag caa ctt gca aga gct tca atg 528Val Ile Lys Arg
Tyr Asp Phe Ala Gln Gln Leu Ala Arg Ala Ser Met 165
170 175cca gaa ttc aag aag aaa ggt tat gaa att
ccc ttg tta gca aca tat 576Pro Glu Phe Lys Lys Lys Gly Tyr Glu Ile
Pro Leu Leu Ala Thr Tyr 180 185
190gca aat gct gca aat gtt cat ttg ctt tta atg aga gat atg caa ata
624Ala Asn Ala Ala Asn Val His Leu Leu Leu Met Arg Asp Met Gln Ile
195 200 205tac ggg gaa aga tgg gga ata
cct aaa gaa gat ata gag ctc tac tta 672Tyr Gly Glu Arg Trp Gly Ile
Pro Lys Glu Asp Ile Glu Leu Tyr Leu 210 215
220tct gaa caa gaa aat ttt acc tct gaa tat aca gat cat tgc gta aaa
720Ser Glu Gln Glu Asn Phe Thr Ser Glu Tyr Thr Asp His Cys Val Lys225
230 235 240tat tat aat gag
gga tta aat caa ttg aaa aat aaa agt ggc gta agt 768Tyr Tyr Asn Glu
Gly Leu Asn Gln Leu Lys Asn Lys Ser Gly Val Ser 245
250 255ggt tta gtt tgg gag aat tat aac cgt ttc
cgt aca gaa atg aca atc 816Gly Leu Val Trp Glu Asn Tyr Asn Arg Phe
Arg Thr Glu Met Thr Ile 260 265
270ctg gtg tta gat att gtg gct gta ttt cca cgc tac aat gta atc gaa
864Leu Val Leu Asp Ile Val Ala Val Phe Pro Arg Tyr Asn Val Ile Glu
275 280 285tat cct ata gat tca aca gta
gaa tta aca aga aca att tat cta gat 912Tyr Pro Ile Asp Ser Thr Val
Glu Leu Thr Arg Thr Ile Tyr Leu Asp 290 295
300cca ctt ggt tac aca ggg aat tcc aat gac gag cat ccc gaa tat tat
960Pro Leu Gly Tyr Thr Gly Asn Ser Asn Asp Glu His Pro Glu Tyr Tyr305
310 315 320gcg tct aca aaa
tca ttt tca tca ata gag agt aga gcc ata cct gca 1008Ala Ser Thr Lys
Ser Phe Ser Ser Ile Glu Ser Arg Ala Ile Pro Ala 325
330 335ccc aca tta ttc cag tgg atc act gaa ctt
caa gta tat tca gca aaa 1056Pro Thr Leu Phe Gln Trp Ile Thr Glu Leu
Gln Val Tyr Ser Ala Lys 340 345
350ggc tct cat ggt tct acc tat act aca tgg tgg act gga cat aaa gtg
1104Gly Ser His Gly Ser Thr Tyr Thr Thr Trp Trp Thr Gly His Lys Val
355 360 365act gct aag cct act aat ggt
ggt ctt gaa agt aaa tat gat ttc gga 1152Thr Ala Lys Pro Thr Asn Gly
Gly Leu Glu Ser Lys Tyr Asp Phe Gly 370 375
380agt tct tca ggt tct cag aac aag gat gtt ttt gct ctt gat ggc aag
1200Ser Ser Ser Gly Ser Gln Asn Lys Asp Val Phe Ala Leu Asp Gly Lys385
390 395 400gat gta tat gat
tca caa agt atg tta aca tcg att agt tat tcc ggt 1248Asp Val Tyr Asp
Ser Gln Ser Met Leu Thr Ser Ile Ser Tyr Ser Gly 405
410 415att aga tat ttt ggg tgt cct cag ttt aag
tta aat tgg ata aat aag 1296Ile Arg Tyr Phe Gly Cys Pro Gln Phe Lys
Leu Asn Trp Ile Asn Lys 420 425
430aat aat gag cta gca gaa cag ata ttt aat tat tca agt aat gtt ggt
1344Asn Asn Glu Leu Ala Glu Gln Ile Phe Asn Tyr Ser Ser Asn Val Gly
435 440 445tca tct ttc agt gag tat agg
tat agc aag gat gaa tta cca ata gaa 1392Ser Ser Phe Ser Glu Tyr Arg
Tyr Ser Lys Asp Glu Leu Pro Ile Glu 450 455
460ttg ttg gcg agc cct att tat gga gat att gag gaa tac agt cat agg
1440Leu Leu Ala Ser Pro Ile Tyr Gly Asp Ile Glu Glu Tyr Ser His Arg465
470 475 480tta agt cac gtt
tca gag gta att aaa gat tat ggg cag gga ata att 1488Leu Ser His Val
Ser Glu Val Ile Lys Asp Tyr Gly Gln Gly Ile Ile 485
490 495cct gta tta ggt ttc aca cat gta agt gta
agt cgt gac aat aga att 1536Pro Val Leu Gly Phe Thr His Val Ser Val
Ser Arg Asp Asn Arg Ile 500 505
510tat tca gat aag att acg caa att cca gct gtg aaa atg tat gaa tta
1584Tyr Ser Asp Lys Ile Thr Gln Ile Pro Ala Val Lys Met Tyr Glu Leu
515 520 525gta agc cca gcc gtt gtt gta
aaa ggg cct gga tct aca ggt gga gat 1632Val Ser Pro Ala Val Val Val
Lys Gly Pro Gly Ser Thr Gly Gly Asp 530 535
540tta gtt aag aga ggg agt agt ggt aat ata gga tct atg aat gtt acc
1680Leu Val Lys Arg Gly Ser Ser Gly Asn Ile Gly Ser Met Asn Val Thr545
550 555 560gta aac tca cca
ctt tct caa aaa tat cgt gtc aga gtt cga tat gcc 1728Val Asn Ser Pro
Leu Ser Gln Lys Tyr Arg Val Arg Val Arg Tyr Ala 565
570 575act aat gct tct ggc cag tta aac gtg agt
att aac gat aaa tta aca 1776Thr Asn Ala Ser Gly Gln Leu Asn Val Ser
Ile Asn Asp Lys Leu Thr 580 585
590ctt caa aaa cct ttt gaa aga acc gga aca aca ata ggt gaa gga aca
1824Leu Gln Lys Pro Phe Glu Arg Thr Gly Thr Thr Ile Gly Glu Gly Thr
595 600 605gat ttg tcc tat gat tca ttt
gga tat tta gaa tat tct acg acg att 1872Asp Leu Ser Tyr Asp Ser Phe
Gly Tyr Leu Glu Tyr Ser Thr Thr Ile 610 615
620caa ttt ccg aat gag cac cca aaa atc act ttt aat tta tcc cat tgg
1920Gln Phe Pro Asn Glu His Pro Lys Ile Thr Phe Asn Leu Ser His Trp625
630 635 640agc ggc agt gga
gca ttt tat ata gat aaa atc gaa ttt atc cct gta 1968Ser Gly Ser Gly
Ala Phe Tyr Ile Asp Lys Ile Glu Phe Ile Pro Val 645
650 655gat gaa aat tac gat gaa aga gta aca cta
gaa aaa gca cag aaa gcc 2016Asp Glu Asn Tyr Asp Glu Arg Val Thr Leu
Glu Lys Ala Gln Lys Ala 660 665
670gtg aat gcc ttg ttt aca gcg gga aga aat gca ctc caa aaa gat gtg
2064Val Asn Ala Leu Phe Thr Ala Gly Arg Asn Ala Leu Gln Lys Asp Val
675 680 685aca gat ttt aaa gta gat cag
gtt tcc att tta gtg gat tgt ata cca 2112Thr Asp Phe Lys Val Asp Gln
Val Ser Ile Leu Val Asp Cys Ile Pro 690 695
700ggg gaa ttg tat cca aat gag aaa cgc gaa cta cta agt tta gtc aaa
2160Gly Glu Leu Tyr Pro Asn Glu Lys Arg Glu Leu Leu Ser Leu Val Lys705
710 715 720tac gca aaa cgg
ttg agt tat tcc cgt aat tta ctc cta gac cca aca 2208Tyr Ala Lys Arg
Leu Ser Tyr Ser Arg Asn Leu Leu Leu Asp Pro Thr 725
730 735ttc gat tct atc aat tca cca gat gag aat
ggc tgg tac ggg agt aat 2256Phe Asp Ser Ile Asn Ser Pro Asp Glu Asn
Gly Trp Tyr Gly Ser Asn 740 745
750ggt att gca att gga aat ggg aac ttt gta ttc aaa gga aac tat tta
2304Gly Ile Ala Ile Gly Asn Gly Asn Phe Val Phe Lys Gly Asn Tyr Leu
755 760 765att ttc tca ggt acc aat gat
aca caa tac cca acg tat ctc tat caa 2352Ile Phe Ser Gly Thr Asn Asp
Thr Gln Tyr Pro Thr Tyr Leu Tyr Gln 770 775
780aaa att gat gaa tcc aag ctc aaa gaa tat aca cgc tat aaa ctg aga
2400Lys Ile Asp Glu Ser Lys Leu Lys Glu Tyr Thr Arg Tyr Lys Leu Arg785
790 795 800ggt ttt atc gag
agt agt cag gat tta gag gca tat gtg att cgc tat 2448Gly Phe Ile Glu
Ser Ser Gln Asp Leu Glu Ala Tyr Val Ile Arg Tyr 805
810 815gat gca aaa cat gaa aca ttg gat gta tcc
aat aat cta ttc cca gat 2496Asp Ala Lys His Glu Thr Leu Asp Val Ser
Asn Asn Leu Phe Pro Asp 820 825
830att tct cct gtg aat gca tgc gga gaa cca aat cgt tgt gcg gca cta
2544Ile Ser Pro Val Asn Ala Cys Gly Glu Pro Asn Arg Cys Ala Ala Leu
835 840 845caa tac ctg gat gaa aac cca
agg tta gaa tgt agt tcg ata caa gat 2592Gln Tyr Leu Asp Glu Asn Pro
Arg Leu Glu Cys Ser Ser Ile Gln Asp 850 855
860ggc att tta tct gat tcg cat tca ttt tct ctc aat ata gat aca ggt
2640Gly Ile Leu Ser Asp Ser His Ser Phe Ser Leu Asn Ile Asp Thr Gly865
870 875 880tct att gat tac
gat gag aac gta ggt att tgg gtg ttg ttt aaa att 2688Ser Ile Asp Tyr
Asp Glu Asn Val Gly Ile Trp Val Leu Phe Lys Ile 885
890 895tcc aca ccg gaa ggg tat gcg aaa ttt gga
aac cta gaa gtg att gaa 2736Ser Thr Pro Glu Gly Tyr Ala Lys Phe Gly
Asn Leu Glu Val Ile Glu 900 905
910gat ggc cta gtc att gga gaa gca tta gcc cgt gtg aaa cgt caa gaa
2784Asp Gly Leu Val Ile Gly Glu Ala Leu Ala Arg Val Lys Arg Gln Glu
915 920 925acg aag tgg aga aac aag ttg
aca caa ctg cga acg gaa aca caa gcg 2832Thr Lys Trp Arg Asn Lys Leu
Thr Gln Leu Arg Thr Glu Thr Gln Ala 930 935
940att tat aca cga gca aaa caa gcc att gat aat tta ttc aca aat gca
2880Ile Tyr Thr Arg Ala Lys Gln Ala Ile Asp Asn Leu Phe Thr Asn Ala945
950 955 960cag gac tct cac
tta aaa ata ggt gct aca ttc gcg tca att gtg gcc 2928Gln Asp Ser His
Leu Lys Ile Gly Ala Thr Phe Ala Ser Ile Val Ala 965
970 975gcg cga aag att gtc caa tca ata cgt gaa
gcg tat atg tca tgg tta 2976Ala Arg Lys Ile Val Gln Ser Ile Arg Glu
Ala Tyr Met Ser Trp Leu 980 985
990tct atc gtc cca agt gta aat tat cct att ttc aca gag ttg aat gaa
3024Ser Ile Val Pro Ser Val Asn Tyr Pro Ile Phe Thr Glu Leu Asn Glu
995 1000 1005aga gta cag cga gca ttt cga
tta tat gat gta cga aat gtc gta cgt 3072Arg Val Gln Arg Ala Phe Arg
Leu Tyr Asp Val Arg Asn Val Val Arg 1010 1015
1020aat ggc cga ttc ttg agt gga gta tcg gat tgg att gtg aca tct gat
3120Asn Gly Arg Phe Leu Ser Gly Val Ser Asp Trp Ile Val Thr Ser
Asp1025 1030 1035 1040gta
aag gta caa gaa gaa aat ggg aac aac gta tta gtt ctt tcc aat 3168Val
Lys Val Gln Glu Glu Asn Gly Asn Asn Val Leu Val Leu Ser Asn
1045 1050 1055tgg gat gca caa gta tta caa
tgt ctg aat ctc tac gaa gac cat ggg 3216Trp Asp Ala Gln Val Leu Gln
Cys Leu Asn Leu Tyr Glu Asp His Gly 1060 1065
1070tat atc tta cgc gta aca gca cgt aag gaa gga ctc gga gaa
gga tat 3264Tyr Ile Leu Arg Val Thr Ala Arg Lys Glu Gly Leu Gly Glu
Gly Tyr 1075 1080 1085ata aca atc
act gat gaa gaa ggg cat aca gat caa ttg aca ttt ggt 3312Ile Thr Ile
Thr Asp Glu Glu Gly His Thr Asp Gln Leu Thr Phe Gly 1090
1095 1100gga tgt gag gag ata gat tca tcc aat tct ttc gta
tct aca ggt tat 3360Gly Cys Glu Glu Ile Asp Ser Ser Asn Ser Phe Val
Ser Thr Gly Tyr1105 1110 1115
1120att aca aaa gag cta gaa ttc ttc cca gat aca gag aaa gtg cgc atc
3408Ile Thr Lys Glu Leu Glu Phe Phe Pro Asp Thr Glu Lys Val Arg Ile
1125 1130 1135gaa att gga gaa aca
gaa gga aca ttc cag gta gaa agt gtg gaa tta 3456Glu Ile Gly Glu Thr
Glu Gly Thr Phe Gln Val Glu Ser Val Glu Leu 1140
1145 1150ttt ttg atg gaa gac ata tgc
3477Phe Leu Met Glu Asp Ile Cys
1155191159PRTBacillus thuringiensis 19Met Asp Ile Arg Asn Gln Lys Lys Ser
Glu Glu Ile His Pro Thr Leu1 5 10
15 Asn Glu Ser Ser Ser Asn Thr Thr Ser Lys Tyr Pro Leu Ala
Ser Asp 20 25 30
Leu Ile Lys Gln Tyr Gln Asn Met Asn Tyr Lys Asp Ser Leu Asn Ile 35
40 45 Ile Glu Glu Asn Asn
Val Ile Asp Pro Val Ser Gly Thr Ala Ala Leu 50 55
60 Val Thr Ala Arg Lys Ile Gly Gly Lys Ile
Ile Lys Lys Leu Gly Glu65 70 75
80 Ala Ala Leu Ser Lys Ile Leu Lys Glu Ile Leu Gly Tyr Leu Trp
Pro 85 90 95 Thr
Ser Gly Glu Asp Asp Ser Trp Thr Asp Met Met Thr Ala Val Glu
100 105 110 Glu Leu Ile Asp Gln
Lys Ile Gln Glu Gln Val Glu Leu Asp Ala Leu 115
120 125 Thr Glu Leu Glu Lys Ile Gly Glu Ala
Val Glu Ala Tyr Tyr Ile Ala 130 135
140 Leu Asp Asp Trp Glu Pro Val Pro Glu Asp Met Phe Ser
Leu Ser Glu145 150 155
160 Val Ile Lys Arg Tyr Asp Phe Ala Gln Gln Leu Ala Arg Ala Ser Met
165 170 175 Pro Glu Phe Lys
Lys Lys Gly Tyr Glu Ile Pro Leu Leu Ala Thr Tyr 180
185 190 Ala Asn Ala Ala Asn Val His Leu Leu
Leu Met Arg Asp Met Gln Ile 195 200
205 Tyr Gly Glu Arg Trp Gly Ile Pro Lys Glu Asp Ile Glu Leu
Tyr Leu 210 215 220
Ser Glu Gln Glu Asn Phe Thr Ser Glu Tyr Thr Asp His Cys Val Lys225
230 235 240 Tyr Tyr Asn Glu Gly
Leu Asn Gln Leu Lys Asn Lys Ser Gly Val Ser 245
250 255 Gly Leu Val Trp Glu Asn Tyr Asn Arg Phe
Arg Thr Glu Met Thr Ile 260 265
270 Leu Val Leu Asp Ile Val Ala Val Phe Pro Arg Tyr Asn Val Ile
Glu 275 280 285 Tyr
Pro Ile Asp Ser Thr Val Glu Leu Thr Arg Thr Ile Tyr Leu Asp 290
295 300 Pro Leu Gly Tyr Thr Gly
Asn Ser Asn Asp Glu His Pro Glu Tyr Tyr305 310
315 320 Ala Ser Thr Lys Ser Phe Ser Ser Ile Glu Ser
Arg Ala Ile Pro Ala 325 330
335 Pro Thr Leu Phe Gln Trp Ile Thr Glu Leu Gln Val Tyr Ser Ala Lys
340 345 350 Gly Ser His
Gly Ser Thr Tyr Thr Thr Trp Trp Thr Gly His Lys Val 355
360 365 Thr Ala Lys Pro Thr Asn Gly Gly
Leu Glu Ser Lys Tyr Asp Phe Gly 370 375
380 Ser Ser Ser Gly Ser Gln Asn Lys Asp Val Phe Ala Leu
Asp Gly Lys385 390 395
400 Asp Val Tyr Asp Ser Gln Ser Met Leu Thr Ser Ile Ser Tyr Ser Gly
405 410 415 Ile Arg Tyr Phe
Gly Cys Pro Gln Phe Lys Leu Asn Trp Ile Asn Lys 420
425 430 Asn Asn Glu Leu Ala Glu Gln Ile Phe
Asn Tyr Ser Ser Asn Val Gly 435 440
445 Ser Ser Phe Ser Glu Tyr Arg Tyr Ser Lys Asp Glu Leu Pro
Ile Glu 450 455 460
Leu Leu Ala Ser Pro Ile Tyr Gly Asp Ile Glu Glu Tyr Ser His Arg465
470 475 480 Leu Ser His Val Ser
Glu Val Ile Lys Asp Tyr Gly Gln Gly Ile Ile 485
490 495 Pro Val Leu Gly Phe Thr His Val Ser Val
Ser Arg Asp Asn Arg Ile 500 505
510 Tyr Ser Asp Lys Ile Thr Gln Ile Pro Ala Val Lys Met Tyr Glu
Leu 515 520 525 Val
Ser Pro Ala Val Val Val Lys Gly Pro Gly Ser Thr Gly Gly Asp 530
535 540 Leu Val Lys Arg Gly Ser
Ser Gly Asn Ile Gly Ser Met Asn Val Thr545 550
555 560 Val Asn Ser Pro Leu Ser Gln Lys Tyr Arg Val
Arg Val Arg Tyr Ala 565 570
575 Thr Asn Ala Ser Gly Gln Leu Asn Val Ser Ile Asn Asp Lys Leu Thr
580 585 590 Leu Gln Lys
Pro Phe Glu Arg Thr Gly Thr Thr Ile Gly Glu Gly Thr 595
600 605 Asp Leu Ser Tyr Asp Ser Phe Gly
Tyr Leu Glu Tyr Ser Thr Thr Ile 610 615
620 Gln Phe Pro Asn Glu His Pro Lys Ile Thr Phe Asn Leu
Ser His Trp625 630 635
640 Ser Gly Ser Gly Ala Phe Tyr Ile Asp Lys Ile Glu Phe Ile Pro Val
645 650 655 Asp Glu Asn Tyr
Asp Glu Arg Val Thr Leu Glu Lys Ala Gln Lys Ala 660
665 670 Val Asn Ala Leu Phe Thr Ala Gly Arg
Asn Ala Leu Gln Lys Asp Val 675 680
685 Thr Asp Phe Lys Val Asp Gln Val Ser Ile Leu Val Asp Cys
Ile Pro 690 695 700
Gly Glu Leu Tyr Pro Asn Glu Lys Arg Glu Leu Leu Ser Leu Val Lys705
710 715 720 Tyr Ala Lys Arg Leu
Ser Tyr Ser Arg Asn Leu Leu Leu Asp Pro Thr 725
730 735 Phe Asp Ser Ile Asn Ser Pro Asp Glu Asn
Gly Trp Tyr Gly Ser Asn 740 745
750 Gly Ile Ala Ile Gly Asn Gly Asn Phe Val Phe Lys Gly Asn Tyr
Leu 755 760 765 Ile
Phe Ser Gly Thr Asn Asp Thr Gln Tyr Pro Thr Tyr Leu Tyr Gln 770
775 780 Lys Ile Asp Glu Ser Lys
Leu Lys Glu Tyr Thr Arg Tyr Lys Leu Arg785 790
795 800 Gly Phe Ile Glu Ser Ser Gln Asp Leu Glu Ala
Tyr Val Ile Arg Tyr 805 810
815 Asp Ala Lys His Glu Thr Leu Asp Val Ser Asn Asn Leu Phe Pro Asp
820 825 830 Ile Ser Pro
Val Asn Ala Cys Gly Glu Pro Asn Arg Cys Ala Ala Leu 835
840 845 Gln Tyr Leu Asp Glu Asn Pro Arg
Leu Glu Cys Ser Ser Ile Gln Asp 850 855
860 Gly Ile Leu Ser Asp Ser His Ser Phe Ser Leu Asn Ile
Asp Thr Gly865 870 875
880 Ser Ile Asp Tyr Asp Glu Asn Val Gly Ile Trp Val Leu Phe Lys Ile
885 890 895 Ser Thr Pro Glu
Gly Tyr Ala Lys Phe Gly Asn Leu Glu Val Ile Glu 900
905 910 Asp Gly Leu Val Ile Gly Glu Ala Leu
Ala Arg Val Lys Arg Gln Glu 915 920
925 Thr Lys Trp Arg Asn Lys Leu Thr Gln Leu Arg Thr Glu Thr
Gln Ala 930 935 940
Ile Tyr Thr Arg Ala Lys Gln Ala Ile Asp Asn Leu Phe Thr Asn Ala945
950 955 960 Gln Asp Ser His Leu
Lys Ile Gly Ala Thr Phe Ala Ser Ile Val Ala 965
970 975 Ala Arg Lys Ile Val Gln Ser Ile Arg Glu
Ala Tyr Met Ser Trp Leu 980 985
990 Ser Ile Val Pro Ser Val Asn Tyr Pro Ile Phe Thr Glu Leu Asn
Glu 995 1000 1005 Arg
Val Gln Arg Ala Phe Arg Leu Tyr Asp Val Arg Asn Val Val Arg 1010
1015 1020 Asn Gly Arg Phe Leu Ser
Gly Val Ser Asp Trp Ile Val Thr Ser Asp1025 1030
1035 1040 Val Lys Val Gln Glu Glu Asn Gly Asn Asn Val
Leu Val Leu Ser Asn 1045 1050
1055 Trp Asp Ala Gln Val Leu Gln Cys Leu Asn Leu Tyr Glu Asp His Gly
1060 1065 1070 Tyr Ile Leu
Arg Val Thr Ala Arg Lys Glu Gly Leu Gly Glu Gly Tyr 1075
1080 1085 Ile Thr Ile Thr Asp Glu Glu Gly
His Thr Asp Gln Leu Thr Phe Gly 1090 1095
1100 Gly Cys Glu Glu Ile Asp Ser Ser Asn Ser Phe Val Ser
Thr Gly Tyr1105 1110 1115
1120 Ile Thr Lys Glu Leu Glu Phe Phe Pro Asp Thr Glu Lys Val Arg Ile
1125 1130 1135 Glu Ile Gly Glu
Thr Glu Gly Thr Phe Gln Val Glu Ser Val Glu Leu 1140
1145 1150 Phe Leu Met Glu Asp Ile Cys
1155 203438DNABacillus thuringiensisCDS(1)...(3438) 20atg
aaa aaa att aag ttt aaa tat tgt ata caa gga gac ttg aat atg 48Met
Lys Lys Ile Lys Phe Lys Tyr Cys Ile Gln Gly Asp Leu Asn Met1
5 10 15aat caa aaa aac tat gat att
ata ggt gct tcg aca aac ggt aca aac 96Asn Gln Lys Asn Tyr Asp Ile
Ile Gly Ala Ser Thr Asn Gly Thr Asn 20 25
30aaa tta ctt gaa ggt tat aac att ata atc agt ccc tac gaa
gct cca 144Lys Leu Leu Glu Gly Tyr Asn Ile Ile Ile Ser Pro Tyr Glu
Ala Pro 35 40 45aca tcc gtt act
aca act att gaa att act gga acc ata cta agc gat 192Thr Ser Val Thr
Thr Thr Ile Glu Ile Thr Gly Thr Ile Leu Ser Asp 50 55
60tta ggt gtt cca gga gca tca tca gtt agt tta ctt ttg
aat aaa ctt 240Leu Gly Val Pro Gly Ala Ser Ser Val Ser Leu Leu Leu
Asn Lys Leu65 70 75
80ata aat cta tta tgg cca aat gat acc aat act gtg tgg ggg aca ttc
288Ile Asn Leu Leu Trp Pro Asn Asp Thr Asn Thr Val Trp Gly Thr Phe
85 90 95gga aaa gaa acc gct gat
ctt cta aat gaa gtg tta tct cca aat gat 336Gly Lys Glu Thr Ala Asp
Leu Leu Asn Glu Val Leu Ser Pro Asn Asp 100
105 110cca gta gta aca gat gca aat act aat tta gca gga
cta aat gac tcc 384Pro Val Val Thr Asp Ala Asn Thr Asn Leu Ala Gly
Leu Asn Asp Ser 115 120 125ctt aac
tta tat tta aat gaa ctt gaa ata tgg aaa aaa gac ccc aac 432Leu Asn
Leu Tyr Leu Asn Glu Leu Glu Ile Trp Lys Lys Asp Pro Asn 130
135 140aac gca act acc caa agg aat gtc aca caa tac
ttt gtt agt ttg aat 480Asn Ala Thr Thr Gln Arg Asn Val Thr Gln Tyr
Phe Val Ser Leu Asn145 150 155
160ttg gat ttt aca cat gat atg cct tca ttt gct gta cct gga tat gaa
528Leu Asp Phe Thr His Asp Met Pro Ser Phe Ala Val Pro Gly Tyr Glu
165 170 175acg aag tta tta aca
att tat gca caa gct gca aat ctt cat tta ctt 576Thr Lys Leu Leu Thr
Ile Tyr Ala Gln Ala Ala Asn Leu His Leu Leu 180
185 190tta tta aga gat gct tct agg ttt gga gaa ggt tgg
gga ctg act caa 624Leu Leu Arg Asp Ala Ser Arg Phe Gly Glu Gly Trp
Gly Leu Thr Gln 195 200 205gaa atc
ata gat tct aac tat aat aat caa tta aaa ttg aca gaa aaa 672Glu Ile
Ile Asp Ser Asn Tyr Asn Asn Gln Leu Lys Leu Thr Glu Lys 210
215 220tac acg gac cat tgt gta aag tgg tac aac gca
gga tta gaa aaa tta 720Tyr Thr Asp His Cys Val Lys Trp Tyr Asn Ala
Gly Leu Glu Lys Leu225 230 235
240aaa gga aat tta act ggg gaa aat tgg tac act tat aat aga ttt cgt
768Lys Gly Asn Leu Thr Gly Glu Asn Trp Tyr Thr Tyr Asn Arg Phe Arg
245 250 255aga gaa atg acg tta
atg gtg tta gac gta gtt gca tta ttt cca aac 816Arg Glu Met Thr Leu
Met Val Leu Asp Val Val Ala Leu Phe Pro Asn 260
265 270tat gat aca cga atg tac ccg atc gca acg tca tca
gaa ctt aca aga 864Tyr Asp Thr Arg Met Tyr Pro Ile Ala Thr Ser Ser
Glu Leu Thr Arg 275 280 285atg att
tat aca gat cca atc gct tat aca caa agc gat cca tgg tac 912Met Ile
Tyr Thr Asp Pro Ile Ala Tyr Thr Gln Ser Asp Pro Trp Tyr 290
295 300aag ata aca tct ctt tct ttt tca aat att gaa
aac agc gcg att cca 960Lys Ile Thr Ser Leu Ser Phe Ser Asn Ile Glu
Asn Ser Ala Ile Pro305 310 315
320agt cct tct ttc ttc aag tgg cta aga tcc gtt tca att aat agc cag
1008Ser Pro Ser Phe Phe Lys Trp Leu Arg Ser Val Ser Ile Asn Ser Gln
325 330 335tgg tgg ggc agt ggt
cct aat caa acc tac tat tgg gtt gga cat gaa 1056Trp Trp Gly Ser Gly
Pro Asn Gln Thr Tyr Tyr Trp Val Gly His Glu 340
345 350tta gta tat tct aat tca aat tct aat caa tca ctt
aag gtt aaa tat 1104Leu Val Tyr Ser Asn Ser Asn Ser Asn Gln Ser Leu
Lys Val Lys Tyr 355 360 365gga gac
cct aat tct tat att gag ccc cct gat tct ttc agt ttt tct 1152Gly Asp
Pro Asn Ser Tyr Ile Glu Pro Pro Asp Ser Phe Ser Phe Ser 370
375 380tct acg gat gtt tac aga acc att tct gtt gtt
aga aat tca gta agt 1200Ser Thr Asp Val Tyr Arg Thr Ile Ser Val Val
Arg Asn Ser Val Ser385 390 395
400aat tat ata gta agt gaa gtt caa ttc aat tca att agt aat aca aat
1248Asn Tyr Ile Val Ser Glu Val Gln Phe Asn Ser Ile Ser Asn Thr Asn
405 410 415caa att agt gaa gaa
att tat aaa cat caa tca aat tgg agt aga aaa 1296Gln Ile Ser Glu Glu
Ile Tyr Lys His Gln Ser Asn Trp Ser Arg Lys 420
425 430gaa acc aaa gat tca att aca gaa cta tcc tta gct
gct aat ccc cca 1344Glu Thr Lys Asp Ser Ile Thr Glu Leu Ser Leu Ala
Ala Asn Pro Pro 435 440 445aca aca
ttt gga aat gta gca gaa tac agt cat aga tta gca tat att 1392Thr Thr
Phe Gly Asn Val Ala Glu Tyr Ser His Arg Leu Ala Tyr Ile 450
455 460tca gag gca tac caa agt caa aac cca tca aaa
tac cca acc tac att 1440Ser Glu Ala Tyr Gln Ser Gln Asn Pro Ser Lys
Tyr Pro Thr Tyr Ile465 470 475
480cct gta ttc ggt tgg acg cat aca agc gta cgt tac gat aat aaa att
1488Pro Val Phe Gly Trp Thr His Thr Ser Val Arg Tyr Asp Asn Lys Ile
485 490 495ttc ccg gac aaa atc
act caa att cca gct gtt aaa agc tct tca gct 1536Phe Pro Asp Lys Ile
Thr Gln Ile Pro Ala Val Lys Ser Ser Ser Ala 500
505 510caa ggt gga tca tgg aaa aat ata gtg aaa ggt ccc
ggg ttt act gga 1584Gln Gly Gly Ser Trp Lys Asn Ile Val Lys Gly Pro
Gly Phe Thr Gly 515 520 525gga gat
gtg aca act gca gtt tcg cca gca act tta acc gac ata ata 1632Gly Asp
Val Thr Thr Ala Val Ser Pro Ala Thr Leu Thr Asp Ile Ile 530
535 540aaa ata caa gtt act cta gat cca aat tca ctt
tca caa aaa tat cgt 1680Lys Ile Gln Val Thr Leu Asp Pro Asn Ser Leu
Ser Gln Lys Tyr Arg545 550 555
560gca cga ctt cgc tat gct tcc aat gca ttt gta gca gct act ttg tat
1728Ala Arg Leu Arg Tyr Ala Ser Asn Ala Phe Val Ala Ala Thr Leu Tyr
565 570 575aca aat aca agt agt
aat tat aat ttt gaa ctt aca aaa ggt aca act 1776Thr Asn Thr Ser Ser
Asn Tyr Asn Phe Glu Leu Thr Lys Gly Thr Thr 580
585 590gaa cag ttt aca aca tat aat tca tac cag tat gta
gat atc cca ggt 1824Glu Gln Phe Thr Thr Tyr Asn Ser Tyr Gln Tyr Val
Asp Ile Pro Gly 595 600 605tca ata
caa ttt aat aat act tct gat aca gtg tct gtt tat ttg cat 1872Ser Ile
Gln Phe Asn Asn Thr Ser Asp Thr Val Ser Val Tyr Leu His 610
615 620atg gat tca aca act aat gca aac gtt cat gta
gat aga att gaa ttc 1920Met Asp Ser Thr Thr Asn Ala Asn Val His Val
Asp Arg Ile Glu Phe625 630 635
640att cca gta gat gaa cag tac gat gaa aga gta aca cta gaa aaa gca
1968Ile Pro Val Asp Glu Gln Tyr Asp Glu Arg Val Thr Leu Glu Lys Ala
645 650 655cag aaa gcc gtg aat
gcc ttg ttt aca gcg gga aga cat gca ctc caa 2016Gln Lys Ala Val Asn
Ala Leu Phe Thr Ala Gly Arg His Ala Leu Gln 660
665 670aca gat gtg aca gat tac aaa gtg gat caa gtg tca
att tta gtg gat 2064Thr Asp Val Thr Asp Tyr Lys Val Asp Gln Val Ser
Ile Leu Val Asp 675 680 685tgt gta
tca ggg gag tta tat cca aat gag aaa cgc gaa cta ctc agt 2112Cys Val
Ser Gly Glu Leu Tyr Pro Asn Glu Lys Arg Glu Leu Leu Ser 690
695 700tta gtc aaa tac gca aaa cgt ttg agc tat tct
cgt aat tta ctc cta 2160Leu Val Lys Tyr Ala Lys Arg Leu Ser Tyr Ser
Arg Asn Leu Leu Leu705 710 715
720gat cca aca ttc gat tct atc aat tca tca gat aag aat ggc tgg tac
2208Asp Pro Thr Phe Asp Ser Ile Asn Ser Ser Asp Lys Asn Gly Trp Tyr
725 730 735ggg agt aat ggt att
gca att agc agt ggg aat ttt gta ttc aaa ggg 2256Gly Ser Asn Gly Ile
Ala Ile Ser Ser Gly Asn Phe Val Phe Lys Gly 740
745 750aac tat tta atc ttc tca ggt aca aat gat gaa caa
tat cca acc tat 2304Asn Tyr Leu Ile Phe Ser Gly Thr Asn Asp Glu Gln
Tyr Pro Thr Tyr 755 760 765ctc tat
caa aaa ata gac gaa tct aag tta aaa gaa tat aca cgt tat 2352Leu Tyr
Gln Lys Ile Asp Glu Ser Lys Leu Lys Glu Tyr Thr Arg Tyr 770
775 780aaa ctg aga ggc ttt ata gaa aat agt caa gat
tta gaa gca tat gtc 2400Lys Leu Arg Gly Phe Ile Glu Asn Ser Gln Asp
Leu Glu Ala Tyr Val785 790 795
800att cgc tat gat gcc aaa cat gaa aca ttg gat gta tcc aat aat cta
2448Ile Arg Tyr Asp Ala Lys His Glu Thr Leu Asp Val Ser Asn Asn Leu
805 810 815tta ccg gat att tct
cct gta aat gca tgt gga gaa cca aat cgt tgt 2496Leu Pro Asp Ile Ser
Pro Val Asn Ala Cys Gly Glu Pro Asn Arg Cys 820
825 830gtg gca tta caa tac ctg gat gaa aac cca aga tta
gaa tgt agt tcg 2544Val Ala Leu Gln Tyr Leu Asp Glu Asn Pro Arg Leu
Glu Cys Ser Ser 835 840 845gtt caa
gat ggt att tta tct gat tcg cat tca ttt tct ctc aat ata 2592Val Gln
Asp Gly Ile Leu Ser Asp Ser His Ser Phe Ser Leu Asn Ile 850
855 860gat aca ggt tct att gat ttc aat gag agc gta
gga att tgg gtg ttg 2640Asp Thr Gly Ser Ile Asp Phe Asn Glu Ser Val
Gly Ile Trp Val Leu865 870 875
880ttt aaa att tcc aca ccg gaa ggg tat gcg aaa ttt gga aac cta gaa
2688Phe Lys Ile Ser Thr Pro Glu Gly Tyr Ala Lys Phe Gly Asn Leu Glu
885 890 895gtg att gaa aat ggc
cca gtc atc gga gaa gca tta gcc cgt gtg aaa 2736Val Ile Glu Asn Gly
Pro Val Ile Gly Glu Ala Leu Ala Arg Val Lys 900
905 910cgc caa gaa aca aag tgg aga aac cag ttg aca caa
ctg aga acg gaa 2784Arg Gln Glu Thr Lys Trp Arg Asn Gln Leu Thr Gln
Leu Arg Thr Glu 915 920 925aca caa
gcg att tat aca cga gca aaa caa gcg ctg gat aat ctt ttt 2832Thr Gln
Ala Ile Tyr Thr Arg Ala Lys Gln Ala Leu Asp Asn Leu Phe 930
935 940gcg aat gca caa gac tct cac tta aaa ata ggt
acg aca ttt gcg gca 2880Ala Asn Ala Gln Asp Ser His Leu Lys Ile Gly
Thr Thr Phe Ala Ala945 950 955
960att gtg gct gcg cga aag att gtc caa tcc ata cgc gaa gcg tat atg
2928Ile Val Ala Ala Arg Lys Ile Val Gln Ser Ile Arg Glu Ala Tyr Met
965 970 975tca tgg tta tct gtt
gtt cca ggt gta aat tat cct atc ttt aca gag 2976Ser Trp Leu Ser Val
Val Pro Gly Val Asn Tyr Pro Ile Phe Thr Glu 980
985 990ttg act gag aga gta cag caa gca ttt caa tta tat
gat gta cga aat 3024Leu Thr Glu Arg Val Gln Gln Ala Phe Gln Leu Tyr
Asp Val Arg Asn 995 1000 1005gtc
gtg cgt aat ggc caa ttc ctt agt ggc tta tcc gat tgg att gta 3072Val
Val Arg Asn Gly Gln Phe Leu Ser Gly Leu Ser Asp Trp Ile Val 1010
1015 1020aca cct gac gtc aag gta caa gaa gac aat
ggg aat aac gta ttg gtt 3120Thr Pro Asp Val Lys Val Gln Glu Asp Asn
Gly Asn Asn Val Leu Val1025 1030 1035
1040ctt tct aat tgg gat gcg caa gta tta caa tgt ctg aag ctc tat
caa 3168Leu Ser Asn Trp Asp Ala Gln Val Leu Gln Cys Leu Lys Leu Tyr
Gln 1045 1050 1055gat cgc
ggg tat atc tta cgt gta acg gca cgt aag gaa gga ttg gga 3216Asp Arg
Gly Tyr Ile Leu Arg Val Thr Ala Arg Lys Glu Gly Leu Gly 1060
1065 1070gaa gga tac gta aca att acg gat gaa
gaa ggg aat aca gat caa ttg 3264Glu Gly Tyr Val Thr Ile Thr Asp Glu
Glu Gly Asn Thr Asp Gln Leu 1075 1080
1085acg ttt ggt gca tgt gag gag ata gat gca tct aat gcg ttc att tcc
3312Thr Phe Gly Ala Cys Glu Glu Ile Asp Ala Ser Asn Ala Phe Ile Ser
1090 1095 1100aca ggt tat att aca aaa gaa
ctg gaa ttc ttc cca gat aca gag aaa 3360Thr Gly Tyr Ile Thr Lys Glu
Leu Glu Phe Phe Pro Asp Thr Glu Lys1105 1110
1115 1120gtg cgt ata gaa att gga gaa aca gaa gga aca ttc
cag gtg gaa agt 3408Val Arg Ile Glu Ile Gly Glu Thr Glu Gly Thr Phe
Gln Val Glu Ser 1125 1130
1135gta gaa tta ttc ttg atg gaa gat cta tgt
3438Val Glu Leu Phe Leu Met Glu Asp Leu Cys 1140
1145211146PRTBacillus thuringiensis 21Met Lys Lys Ile Lys Phe Lys Tyr
Cys Ile Gln Gly Asp Leu Asn Met1 5 10
15 Asn Gln Lys Asn Tyr Asp Ile Ile Gly Ala Ser Thr Asn
Gly Thr Asn 20 25 30
Lys Leu Leu Glu Gly Tyr Asn Ile Ile Ile Ser Pro Tyr Glu Ala Pro
35 40 45 Thr Ser Val Thr
Thr Thr Ile Glu Ile Thr Gly Thr Ile Leu Ser Asp 50 55
60 Leu Gly Val Pro Gly Ala Ser Ser Val
Ser Leu Leu Leu Asn Lys Leu65 70 75
80 Ile Asn Leu Leu Trp Pro Asn Asp Thr Asn Thr Val Trp Gly
Thr Phe 85 90 95
Gly Lys Glu Thr Ala Asp Leu Leu Asn Glu Val Leu Ser Pro Asn Asp
100 105 110 Pro Val Val Thr Asp
Ala Asn Thr Asn Leu Ala Gly Leu Asn Asp Ser 115
120 125 Leu Asn Leu Tyr Leu Asn Glu Leu Glu
Ile Trp Lys Lys Asp Pro Asn 130 135
140 Asn Ala Thr Thr Gln Arg Asn Val Thr Gln Tyr Phe Val
Ser Leu Asn145 150 155
160 Leu Asp Phe Thr His Asp Met Pro Ser Phe Ala Val Pro Gly Tyr Glu
165 170 175 Thr Lys Leu Leu
Thr Ile Tyr Ala Gln Ala Ala Asn Leu His Leu Leu 180
185 190 Leu Leu Arg Asp Ala Ser Arg Phe Gly
Glu Gly Trp Gly Leu Thr Gln 195 200
205 Glu Ile Ile Asp Ser Asn Tyr Asn Asn Gln Leu Lys Leu Thr
Glu Lys 210 215 220
Tyr Thr Asp His Cys Val Lys Trp Tyr Asn Ala Gly Leu Glu Lys Leu225
230 235 240 Lys Gly Asn Leu Thr
Gly Glu Asn Trp Tyr Thr Tyr Asn Arg Phe Arg 245
250 255 Arg Glu Met Thr Leu Met Val Leu Asp Val
Val Ala Leu Phe Pro Asn 260 265
270 Tyr Asp Thr Arg Met Tyr Pro Ile Ala Thr Ser Ser Glu Leu Thr
Arg 275 280 285 Met
Ile Tyr Thr Asp Pro Ile Ala Tyr Thr Gln Ser Asp Pro Trp Tyr 290
295 300 Lys Ile Thr Ser Leu Ser
Phe Ser Asn Ile Glu Asn Ser Ala Ile Pro305 310
315 320 Ser Pro Ser Phe Phe Lys Trp Leu Arg Ser Val
Ser Ile Asn Ser Gln 325 330
335 Trp Trp Gly Ser Gly Pro Asn Gln Thr Tyr Tyr Trp Val Gly His Glu
340 345 350 Leu Val Tyr
Ser Asn Ser Asn Ser Asn Gln Ser Leu Lys Val Lys Tyr 355
360 365 Gly Asp Pro Asn Ser Tyr Ile Glu
Pro Pro Asp Ser Phe Ser Phe Ser 370 375
380 Ser Thr Asp Val Tyr Arg Thr Ile Ser Val Val Arg Asn
Ser Val Ser385 390 395
400 Asn Tyr Ile Val Ser Glu Val Gln Phe Asn Ser Ile Ser Asn Thr Asn
405 410 415 Gln Ile Ser Glu
Glu Ile Tyr Lys His Gln Ser Asn Trp Ser Arg Lys 420
425 430 Glu Thr Lys Asp Ser Ile Thr Glu Leu
Ser Leu Ala Ala Asn Pro Pro 435 440
445 Thr Thr Phe Gly Asn Val Ala Glu Tyr Ser His Arg Leu Ala
Tyr Ile 450 455 460
Ser Glu Ala Tyr Gln Ser Gln Asn Pro Ser Lys Tyr Pro Thr Tyr Ile465
470 475 480 Pro Val Phe Gly Trp
Thr His Thr Ser Val Arg Tyr Asp Asn Lys Ile 485
490 495 Phe Pro Asp Lys Ile Thr Gln Ile Pro Ala
Val Lys Ser Ser Ser Ala 500 505
510 Gln Gly Gly Ser Trp Lys Asn Ile Val Lys Gly Pro Gly Phe Thr
Gly 515 520 525 Gly
Asp Val Thr Thr Ala Val Ser Pro Ala Thr Leu Thr Asp Ile Ile 530
535 540 Lys Ile Gln Val Thr Leu
Asp Pro Asn Ser Leu Ser Gln Lys Tyr Arg545 550
555 560 Ala Arg Leu Arg Tyr Ala Ser Asn Ala Phe Val
Ala Ala Thr Leu Tyr 565 570
575 Thr Asn Thr Ser Ser Asn Tyr Asn Phe Glu Leu Thr Lys Gly Thr Thr
580 585 590 Glu Gln Phe
Thr Thr Tyr Asn Ser Tyr Gln Tyr Val Asp Ile Pro Gly 595
600 605 Ser Ile Gln Phe Asn Asn Thr Ser
Asp Thr Val Ser Val Tyr Leu His 610 615
620 Met Asp Ser Thr Thr Asn Ala Asn Val His Val Asp Arg
Ile Glu Phe625 630 635
640 Ile Pro Val Asp Glu Gln Tyr Asp Glu Arg Val Thr Leu Glu Lys Ala
645 650 655 Gln Lys Ala Val
Asn Ala Leu Phe Thr Ala Gly Arg His Ala Leu Gln 660
665 670 Thr Asp Val Thr Asp Tyr Lys Val Asp
Gln Val Ser Ile Leu Val Asp 675 680
685 Cys Val Ser Gly Glu Leu Tyr Pro Asn Glu Lys Arg Glu Leu
Leu Ser 690 695 700
Leu Val Lys Tyr Ala Lys Arg Leu Ser Tyr Ser Arg Asn Leu Leu Leu705
710 715 720 Asp Pro Thr Phe Asp
Ser Ile Asn Ser Ser Asp Lys Asn Gly Trp Tyr 725
730 735 Gly Ser Asn Gly Ile Ala Ile Ser Ser Gly
Asn Phe Val Phe Lys Gly 740 745
750 Asn Tyr Leu Ile Phe Ser Gly Thr Asn Asp Glu Gln Tyr Pro Thr
Tyr 755 760 765 Leu
Tyr Gln Lys Ile Asp Glu Ser Lys Leu Lys Glu Tyr Thr Arg Tyr 770
775 780 Lys Leu Arg Gly Phe Ile
Glu Asn Ser Gln Asp Leu Glu Ala Tyr Val785 790
795 800 Ile Arg Tyr Asp Ala Lys His Glu Thr Leu Asp
Val Ser Asn Asn Leu 805 810
815 Leu Pro Asp Ile Ser Pro Val Asn Ala Cys Gly Glu Pro Asn Arg Cys
820 825 830 Val Ala Leu
Gln Tyr Leu Asp Glu Asn Pro Arg Leu Glu Cys Ser Ser 835
840 845 Val Gln Asp Gly Ile Leu Ser Asp
Ser His Ser Phe Ser Leu Asn Ile 850 855
860 Asp Thr Gly Ser Ile Asp Phe Asn Glu Ser Val Gly Ile
Trp Val Leu865 870 875
880 Phe Lys Ile Ser Thr Pro Glu Gly Tyr Ala Lys Phe Gly Asn Leu Glu
885 890 895 Val Ile Glu Asn
Gly Pro Val Ile Gly Glu Ala Leu Ala Arg Val Lys 900
905 910 Arg Gln Glu Thr Lys Trp Arg Asn Gln
Leu Thr Gln Leu Arg Thr Glu 915 920
925 Thr Gln Ala Ile Tyr Thr Arg Ala Lys Gln Ala Leu Asp Asn
Leu Phe 930 935 940
Ala Asn Ala Gln Asp Ser His Leu Lys Ile Gly Thr Thr Phe Ala Ala945
950 955 960 Ile Val Ala Ala Arg
Lys Ile Val Gln Ser Ile Arg Glu Ala Tyr Met 965
970 975 Ser Trp Leu Ser Val Val Pro Gly Val Asn
Tyr Pro Ile Phe Thr Glu 980 985
990 Leu Thr Glu Arg Val Gln Gln Ala Phe Gln Leu Tyr Asp Val Arg
Asn 995 1000 1005 Val
Val Arg Asn Gly Gln Phe Leu Ser Gly Leu Ser Asp Trp Ile Val 1010
1015 1020 Thr Pro Asp Val Lys Val
Gln Glu Asp Asn Gly Asn Asn Val Leu Val1025 1030
1035 1040 Leu Ser Asn Trp Asp Ala Gln Val Leu Gln Cys
Leu Lys Leu Tyr Gln 1045 1050
1055 Asp Arg Gly Tyr Ile Leu Arg Val Thr Ala Arg Lys Glu Gly Leu Gly
1060 1065 1070 Glu Gly Tyr
Val Thr Ile Thr Asp Glu Glu Gly Asn Thr Asp Gln Leu 1075
1080 1085 Thr Phe Gly Ala Cys Glu Glu Ile
Asp Ala Ser Asn Ala Phe Ile Ser 1090 1095
1100 Thr Gly Tyr Ile Thr Lys Glu Leu Glu Phe Phe Pro Asp
Thr Glu Lys1105 1110 1115
1120 Val Arg Ile Glu Ile Gly Glu Thr Glu Gly Thr Phe Gln Val Glu Ser
1125 1130 1135 Val Glu Leu Phe
Leu Met Glu Asp Leu Cys 1140 1145
223393DNABacillus thuringiensisCDS(1)...(3393) 22atg aat caa aaa aac tat
gat att ata ggt gct tcg aca aac ggt aca 48Met Asn Gln Lys Asn Tyr
Asp Ile Ile Gly Ala Ser Thr Asn Gly Thr1 5
10 15aac aaa tta ctt gaa ggt tat aac att ata atc agt
ccc tac gaa gct 96Asn Lys Leu Leu Glu Gly Tyr Asn Ile Ile Ile Ser
Pro Tyr Glu Ala 20 25 30cca
aca tcc gtt act aca act att gaa att act gga acc ata cta agc 144Pro
Thr Ser Val Thr Thr Thr Ile Glu Ile Thr Gly Thr Ile Leu Ser 35
40 45gat tta ggt gtt cca gga gca tca tca
gtt agt tta ctt ttg aat aaa 192Asp Leu Gly Val Pro Gly Ala Ser Ser
Val Ser Leu Leu Leu Asn Lys 50 55
60ctt ata aat cta tta tgg cca aat gat acc aat act gtg tgg ggg aca
240Leu Ile Asn Leu Leu Trp Pro Asn Asp Thr Asn Thr Val Trp Gly Thr65
70 75 80ttc gga aaa gaa acc
gct gat ctt cta aat gaa gtg tta tct cca aat 288Phe Gly Lys Glu Thr
Ala Asp Leu Leu Asn Glu Val Leu Ser Pro Asn 85
90 95gat cca gta gta aca gat gca aat act aat tta
gca gga cta aat gac 336Asp Pro Val Val Thr Asp Ala Asn Thr Asn Leu
Ala Gly Leu Asn Asp 100 105
110tcc ctt aac tta tat tta aat gaa ctt gaa ata tgg aaa aaa gac ccc
384Ser Leu Asn Leu Tyr Leu Asn Glu Leu Glu Ile Trp Lys Lys Asp Pro
115 120 125aac aac gca act acc caa agg
aat gtc aca caa tac ttt gtt agt ttg 432Asn Asn Ala Thr Thr Gln Arg
Asn Val Thr Gln Tyr Phe Val Ser Leu 130 135
140aat ttg gat ttt aca cat gat atg cct tca ttt gct gta cct gga tat
480Asn Leu Asp Phe Thr His Asp Met Pro Ser Phe Ala Val Pro Gly Tyr145
150 155 160gaa acg aag tta
tta aca att tat gca caa gct gca aat ctt cat tta 528Glu Thr Lys Leu
Leu Thr Ile Tyr Ala Gln Ala Ala Asn Leu His Leu 165
170 175ctt tta tta aga gat gct tct agg ttt gga
gaa ggt tgg gga ctg act 576Leu Leu Leu Arg Asp Ala Ser Arg Phe Gly
Glu Gly Trp Gly Leu Thr 180 185
190caa gaa atc ata gat tct aac tat aat aat caa tta aaa ttg aca gaa
624Gln Glu Ile Ile Asp Ser Asn Tyr Asn Asn Gln Leu Lys Leu Thr Glu
195 200 205aaa tac acg gac cat tgt gta
aag tgg tac aac gca gga tta gaa aaa 672Lys Tyr Thr Asp His Cys Val
Lys Trp Tyr Asn Ala Gly Leu Glu Lys 210 215
220tta aaa gga aat tta act ggg gaa aat tgg tac act tat aat aga ttt
720Leu Lys Gly Asn Leu Thr Gly Glu Asn Trp Tyr Thr Tyr Asn Arg Phe225
230 235 240cgt aga gaa atg
acg tta atg gtg tta gac gta gtt gca tta ttt cca 768Arg Arg Glu Met
Thr Leu Met Val Leu Asp Val Val Ala Leu Phe Pro 245
250 255aac tat gat aca cga atg tac ccg atc gca
acg tca tca gaa ctt aca 816Asn Tyr Asp Thr Arg Met Tyr Pro Ile Ala
Thr Ser Ser Glu Leu Thr 260 265
270aga atg att tat aca gat cca atc gct tat aca caa agc gat cca tgg
864Arg Met Ile Tyr Thr Asp Pro Ile Ala Tyr Thr Gln Ser Asp Pro Trp
275 280 285tac aag ata aca tct ctt tct
ttt tca aat att gaa aac agc gcg att 912Tyr Lys Ile Thr Ser Leu Ser
Phe Ser Asn Ile Glu Asn Ser Ala Ile 290 295
300cca agt cct tct ttc ttc aag tgg cta aga tcc gtt tca att aat agc
960Pro Ser Pro Ser Phe Phe Lys Trp Leu Arg Ser Val Ser Ile Asn Ser305
310 315 320cag tgg tgg ggc
agt ggt cct aat caa acc tac tat tgg gtt gga cat 1008Gln Trp Trp Gly
Ser Gly Pro Asn Gln Thr Tyr Tyr Trp Val Gly His 325
330 335gaa tta gta tat tct aat tca aat tct aat
caa tca ctt aag gtt aaa 1056Glu Leu Val Tyr Ser Asn Ser Asn Ser Asn
Gln Ser Leu Lys Val Lys 340 345
350tat gga gac cct aat tct tat att gag ccc cct gat tct ttc agt ttt
1104Tyr Gly Asp Pro Asn Ser Tyr Ile Glu Pro Pro Asp Ser Phe Ser Phe
355 360 365tct tct acg gat gtt tac aga
acc att tct gtt gtt aga aat tca gta 1152Ser Ser Thr Asp Val Tyr Arg
Thr Ile Ser Val Val Arg Asn Ser Val 370 375
380agt aat tat ata gta agt gaa gtt caa ttc aat tca att agt aat aca
1200Ser Asn Tyr Ile Val Ser Glu Val Gln Phe Asn Ser Ile Ser Asn Thr385
390 395 400aat caa att agt
gaa gaa att tat aaa cat caa tca aat tgg agt aga 1248Asn Gln Ile Ser
Glu Glu Ile Tyr Lys His Gln Ser Asn Trp Ser Arg 405
410 415aaa gaa acc aaa gat tca att aca gaa cta
tcc tta gct gct aat ccc 1296Lys Glu Thr Lys Asp Ser Ile Thr Glu Leu
Ser Leu Ala Ala Asn Pro 420 425
430cca aca aca ttt gga aat gta gca gaa tac agt cat aga tta gca tat
1344Pro Thr Thr Phe Gly Asn Val Ala Glu Tyr Ser His Arg Leu Ala Tyr
435 440 445att tca gag gca tac caa agt
caa aac cca tca aaa tac cca acc tac 1392Ile Ser Glu Ala Tyr Gln Ser
Gln Asn Pro Ser Lys Tyr Pro Thr Tyr 450 455
460att cct gta ttc ggt tgg acg cat aca agc gta cgt tac gat aat aaa
1440Ile Pro Val Phe Gly Trp Thr His Thr Ser Val Arg Tyr Asp Asn Lys465
470 475 480att ttc ccg gac
aaa atc act caa att cca gct gtt aaa agc tct tca 1488Ile Phe Pro Asp
Lys Ile Thr Gln Ile Pro Ala Val Lys Ser Ser Ser 485
490 495gct caa ggt gga tca tgg aaa aat ata gtg
aaa ggt ccc ggg ttt act 1536Ala Gln Gly Gly Ser Trp Lys Asn Ile Val
Lys Gly Pro Gly Phe Thr 500 505
510gga gga gat gtg aca act gca gtt tcg cca gca act tta acc gac ata
1584Gly Gly Asp Val Thr Thr Ala Val Ser Pro Ala Thr Leu Thr Asp Ile
515 520 525ata aaa ata caa gtt act cta
gat cca aat tca ctt tca caa aaa tat 1632Ile Lys Ile Gln Val Thr Leu
Asp Pro Asn Ser Leu Ser Gln Lys Tyr 530 535
540cgt gca cga ctt cgc tat gct tcc aat gca ttt gta gca gct act ttg
1680Arg Ala Arg Leu Arg Tyr Ala Ser Asn Ala Phe Val Ala Ala Thr Leu545
550 555 560tat aca aat aca
agt agt aat tat aat ttt gaa ctt aca aaa ggt aca 1728Tyr Thr Asn Thr
Ser Ser Asn Tyr Asn Phe Glu Leu Thr Lys Gly Thr 565
570 575act gaa cag ttt aca aca tat aat tca tac
cag tat gta gat atc cca 1776Thr Glu Gln Phe Thr Thr Tyr Asn Ser Tyr
Gln Tyr Val Asp Ile Pro 580 585
590ggt tca ata caa ttt aat aat act tct gat aca gtg tct gtt tat ttg
1824Gly Ser Ile Gln Phe Asn Asn Thr Ser Asp Thr Val Ser Val Tyr Leu
595 600 605cat atg gat tca aca act aat
gca aac gtt cat gta gat aga att gaa 1872His Met Asp Ser Thr Thr Asn
Ala Asn Val His Val Asp Arg Ile Glu 610 615
620ttc att cca gta gat gaa cag tac gat gaa aga gta aca cta gaa aaa
1920Phe Ile Pro Val Asp Glu Gln Tyr Asp Glu Arg Val Thr Leu Glu Lys625
630 635 640gca cag aaa gcc
gtg aat gcc ttg ttt aca gcg gga aga cat gca ctc 1968Ala Gln Lys Ala
Val Asn Ala Leu Phe Thr Ala Gly Arg His Ala Leu 645
650 655caa aca gat gtg aca gat tac aaa gtg gat
caa gtg tca att tta gtg 2016Gln Thr Asp Val Thr Asp Tyr Lys Val Asp
Gln Val Ser Ile Leu Val 660 665
670gat tgt gta tca ggg gag tta tat cca aat gag aaa cgc gaa cta ctc
2064Asp Cys Val Ser Gly Glu Leu Tyr Pro Asn Glu Lys Arg Glu Leu Leu
675 680 685agt tta gtc aaa tac gca aaa
cgt ttg agc tat tct cgt aat tta ctc 2112Ser Leu Val Lys Tyr Ala Lys
Arg Leu Ser Tyr Ser Arg Asn Leu Leu 690 695
700cta gat cca aca ttc gat tct atc aat tca tca gat aag aat ggc tgg
2160Leu Asp Pro Thr Phe Asp Ser Ile Asn Ser Ser Asp Lys Asn Gly Trp705
710 715 720tac ggg agt aat
ggt att gca att agc agt ggg aat ttt gta ttc aaa 2208Tyr Gly Ser Asn
Gly Ile Ala Ile Ser Ser Gly Asn Phe Val Phe Lys 725
730 735ggg aac tat tta atc ttc tca ggt aca aat
gat gaa caa tat cca acc 2256Gly Asn Tyr Leu Ile Phe Ser Gly Thr Asn
Asp Glu Gln Tyr Pro Thr 740 745
750tat ctc tat caa aaa ata gac gaa tct aag tta aaa gaa tat aca cgt
2304Tyr Leu Tyr Gln Lys Ile Asp Glu Ser Lys Leu Lys Glu Tyr Thr Arg
755 760 765tat aaa ctg aga ggc ttt ata
gaa aat agt caa gat tta gaa gca tat 2352Tyr Lys Leu Arg Gly Phe Ile
Glu Asn Ser Gln Asp Leu Glu Ala Tyr 770 775
780gtc att cgc tat gat gcc aaa cat gaa aca ttg gat gta tcc aat aat
2400Val Ile Arg Tyr Asp Ala Lys His Glu Thr Leu Asp Val Ser Asn Asn785
790 795 800cta tta ccg gat
att tct cct gta aat gca tgt gga gaa cca aat cgt 2448Leu Leu Pro Asp
Ile Ser Pro Val Asn Ala Cys Gly Glu Pro Asn Arg 805
810 815tgt gtg gca tta caa tac ctg gat gaa aac
cca aga tta gaa tgt agt 2496Cys Val Ala Leu Gln Tyr Leu Asp Glu Asn
Pro Arg Leu Glu Cys Ser 820 825
830tcg gtt caa gat ggt att tta tct gat tcg cat tca ttt tct ctc aat
2544Ser Val Gln Asp Gly Ile Leu Ser Asp Ser His Ser Phe Ser Leu Asn
835 840 845ata gat aca ggt tct att gat
ttc aat gag agc gta gga att tgg gtg 2592Ile Asp Thr Gly Ser Ile Asp
Phe Asn Glu Ser Val Gly Ile Trp Val 850 855
860ttg ttt aaa att tcc aca ccg gaa ggg tat gcg aaa ttt gga aac cta
2640Leu Phe Lys Ile Ser Thr Pro Glu Gly Tyr Ala Lys Phe Gly Asn Leu865
870 875 880gaa gtg att gaa
aat ggc cca gtc atc gga gaa gca tta gcc cgt gtg 2688Glu Val Ile Glu
Asn Gly Pro Val Ile Gly Glu Ala Leu Ala Arg Val 885
890 895aaa cgc caa gaa aca aag tgg aga aac cag
ttg aca caa ctg aga acg 2736Lys Arg Gln Glu Thr Lys Trp Arg Asn Gln
Leu Thr Gln Leu Arg Thr 900 905
910gaa aca caa gcg att tat aca cga gca aaa caa gcg ctg gat aat ctt
2784Glu Thr Gln Ala Ile Tyr Thr Arg Ala Lys Gln Ala Leu Asp Asn Leu
915 920 925ttt gcg aat gca caa gac tct
cac tta aaa ata ggt acg aca ttt gcg 2832Phe Ala Asn Ala Gln Asp Ser
His Leu Lys Ile Gly Thr Thr Phe Ala 930 935
940gca att gtg gct gcg cga aag att gtc caa tcc ata cgc gaa gcg tat
2880Ala Ile Val Ala Ala Arg Lys Ile Val Gln Ser Ile Arg Glu Ala Tyr945
950 955 960atg tca tgg tta
tct gtt gtt cca ggt gta aat tat cct atc ttt aca 2928Met Ser Trp Leu
Ser Val Val Pro Gly Val Asn Tyr Pro Ile Phe Thr 965
970 975gag ttg act gag aga gta cag caa gca ttt
caa tta tat gat gta cga 2976Glu Leu Thr Glu Arg Val Gln Gln Ala Phe
Gln Leu Tyr Asp Val Arg 980 985
990aat gtc gtg cgt aat ggc caa ttc ctt agt ggc tta tcc gat tgg att
3024Asn Val Val Arg Asn Gly Gln Phe Leu Ser Gly Leu Ser Asp Trp Ile
995 1000 1005gta aca cct gac gtc aag gta
caa gaa gac aat ggg aat aac gta ttg 3072Val Thr Pro Asp Val Lys Val
Gln Glu Asp Asn Gly Asn Asn Val Leu 1010 1015
1020gtt ctt tct aat tgg gat gcg caa gta tta caa tgt ctg aag ctc tat
3120Val Leu Ser Asn Trp Asp Ala Gln Val Leu Gln Cys Leu Lys Leu
Tyr1025 1030 1035 1040caa
gat cgc ggg tat atc tta cgt gta acg gca cgt aag gaa gga ttg 3168Gln
Asp Arg Gly Tyr Ile Leu Arg Val Thr Ala Arg Lys Glu Gly Leu
1045 1050 1055gga gaa gga tac gta aca att
acg gat gaa gaa ggg aat aca gat caa 3216Gly Glu Gly Tyr Val Thr Ile
Thr Asp Glu Glu Gly Asn Thr Asp Gln 1060 1065
1070ttg acg ttt ggt gca tgt gag gag ata gat gca tct aat gcg
ttc att 3264Leu Thr Phe Gly Ala Cys Glu Glu Ile Asp Ala Ser Asn Ala
Phe Ile 1075 1080 1085tcc aca ggt
tat att aca aaa gaa ctg gaa ttc ttc cca gat aca gag 3312Ser Thr Gly
Tyr Ile Thr Lys Glu Leu Glu Phe Phe Pro Asp Thr Glu 1090
1095 1100aaa gtg cgt ata gaa att gga gaa aca gaa gga aca
ttc cag gtg gaa 3360Lys Val Arg Ile Glu Ile Gly Glu Thr Glu Gly Thr
Phe Gln Val Glu1105 1110 1115
1120agt gta gaa tta ttc ttg atg gaa gat cta tgt
3393Ser Val Glu Leu Phe Leu Met Glu Asp Leu Cys 1125
1130231131PRTBacillus thuringiensis 23Met Asn Gln Lys Asn Tyr
Asp Ile Ile Gly Ala Ser Thr Asn Gly Thr1 5
10 15 Asn Lys Leu Leu Glu Gly Tyr Asn Ile Ile Ile
Ser Pro Tyr Glu Ala 20 25 30
Pro Thr Ser Val Thr Thr Thr Ile Glu Ile Thr Gly Thr Ile Leu Ser
35 40 45 Asp Leu Gly
Val Pro Gly Ala Ser Ser Val Ser Leu Leu Leu Asn Lys 50
55 60 Leu Ile Asn Leu Leu Trp Pro Asn
Asp Thr Asn Thr Val Trp Gly Thr65 70 75
80 Phe Gly Lys Glu Thr Ala Asp Leu Leu Asn Glu Val Leu
Ser Pro Asn 85 90 95
Asp Pro Val Val Thr Asp Ala Asn Thr Asn Leu Ala Gly Leu Asn Asp
100 105 110 Ser Leu Asn Leu Tyr
Leu Asn Glu Leu Glu Ile Trp Lys Lys Asp Pro 115
120 125 Asn Asn Ala Thr Thr Gln Arg Asn Val
Thr Gln Tyr Phe Val Ser Leu 130 135
140 Asn Leu Asp Phe Thr His Asp Met Pro Ser Phe Ala Val
Pro Gly Tyr145 150 155
160 Glu Thr Lys Leu Leu Thr Ile Tyr Ala Gln Ala Ala Asn Leu His Leu
165 170 175 Leu Leu Leu Arg
Asp Ala Ser Arg Phe Gly Glu Gly Trp Gly Leu Thr 180
185 190 Gln Glu Ile Ile Asp Ser Asn Tyr Asn
Asn Gln Leu Lys Leu Thr Glu 195 200
205 Lys Tyr Thr Asp His Cys Val Lys Trp Tyr Asn Ala Gly Leu
Glu Lys 210 215 220
Leu Lys Gly Asn Leu Thr Gly Glu Asn Trp Tyr Thr Tyr Asn Arg Phe225
230 235 240 Arg Arg Glu Met Thr
Leu Met Val Leu Asp Val Val Ala Leu Phe Pro 245
250 255 Asn Tyr Asp Thr Arg Met Tyr Pro Ile Ala
Thr Ser Ser Glu Leu Thr 260 265
270 Arg Met Ile Tyr Thr Asp Pro Ile Ala Tyr Thr Gln Ser Asp Pro
Trp 275 280 285 Tyr
Lys Ile Thr Ser Leu Ser Phe Ser Asn Ile Glu Asn Ser Ala Ile 290
295 300 Pro Ser Pro Ser Phe Phe
Lys Trp Leu Arg Ser Val Ser Ile Asn Ser305 310
315 320 Gln Trp Trp Gly Ser Gly Pro Asn Gln Thr Tyr
Tyr Trp Val Gly His 325 330
335 Glu Leu Val Tyr Ser Asn Ser Asn Ser Asn Gln Ser Leu Lys Val Lys
340 345 350 Tyr Gly Asp
Pro Asn Ser Tyr Ile Glu Pro Pro Asp Ser Phe Ser Phe 355
360 365 Ser Ser Thr Asp Val Tyr Arg Thr
Ile Ser Val Val Arg Asn Ser Val 370 375
380 Ser Asn Tyr Ile Val Ser Glu Val Gln Phe Asn Ser Ile
Ser Asn Thr385 390 395
400 Asn Gln Ile Ser Glu Glu Ile Tyr Lys His Gln Ser Asn Trp Ser Arg
405 410 415 Lys Glu Thr Lys
Asp Ser Ile Thr Glu Leu Ser Leu Ala Ala Asn Pro 420
425 430 Pro Thr Thr Phe Gly Asn Val Ala Glu
Tyr Ser His Arg Leu Ala Tyr 435 440
445 Ile Ser Glu Ala Tyr Gln Ser Gln Asn Pro Ser Lys Tyr Pro
Thr Tyr 450 455 460
Ile Pro Val Phe Gly Trp Thr His Thr Ser Val Arg Tyr Asp Asn Lys465
470 475 480 Ile Phe Pro Asp Lys
Ile Thr Gln Ile Pro Ala Val Lys Ser Ser Ser 485
490 495 Ala Gln Gly Gly Ser Trp Lys Asn Ile Val
Lys Gly Pro Gly Phe Thr 500 505
510 Gly Gly Asp Val Thr Thr Ala Val Ser Pro Ala Thr Leu Thr Asp
Ile 515 520 525 Ile
Lys Ile Gln Val Thr Leu Asp Pro Asn Ser Leu Ser Gln Lys Tyr 530
535 540 Arg Ala Arg Leu Arg Tyr
Ala Ser Asn Ala Phe Val Ala Ala Thr Leu545 550
555 560 Tyr Thr Asn Thr Ser Ser Asn Tyr Asn Phe Glu
Leu Thr Lys Gly Thr 565 570
575 Thr Glu Gln Phe Thr Thr Tyr Asn Ser Tyr Gln Tyr Val Asp Ile Pro
580 585 590 Gly Ser Ile
Gln Phe Asn Asn Thr Ser Asp Thr Val Ser Val Tyr Leu 595
600 605 His Met Asp Ser Thr Thr Asn Ala
Asn Val His Val Asp Arg Ile Glu 610 615
620 Phe Ile Pro Val Asp Glu Gln Tyr Asp Glu Arg Val Thr
Leu Glu Lys625 630 635
640 Ala Gln Lys Ala Val Asn Ala Leu Phe Thr Ala Gly Arg His Ala Leu
645 650 655 Gln Thr Asp Val
Thr Asp Tyr Lys Val Asp Gln Val Ser Ile Leu Val 660
665 670 Asp Cys Val Ser Gly Glu Leu Tyr Pro
Asn Glu Lys Arg Glu Leu Leu 675 680
685 Ser Leu Val Lys Tyr Ala Lys Arg Leu Ser Tyr Ser Arg Asn
Leu Leu 690 695 700
Leu Asp Pro Thr Phe Asp Ser Ile Asn Ser Ser Asp Lys Asn Gly Trp705
710 715 720 Tyr Gly Ser Asn Gly
Ile Ala Ile Ser Ser Gly Asn Phe Val Phe Lys 725
730 735 Gly Asn Tyr Leu Ile Phe Ser Gly Thr Asn
Asp Glu Gln Tyr Pro Thr 740 745
750 Tyr Leu Tyr Gln Lys Ile Asp Glu Ser Lys Leu Lys Glu Tyr Thr
Arg 755 760 765 Tyr
Lys Leu Arg Gly Phe Ile Glu Asn Ser Gln Asp Leu Glu Ala Tyr 770
775 780 Val Ile Arg Tyr Asp Ala
Lys His Glu Thr Leu Asp Val Ser Asn Asn785 790
795 800 Leu Leu Pro Asp Ile Ser Pro Val Asn Ala Cys
Gly Glu Pro Asn Arg 805 810
815 Cys Val Ala Leu Gln Tyr Leu Asp Glu Asn Pro Arg Leu Glu Cys Ser
820 825 830 Ser Val Gln
Asp Gly Ile Leu Ser Asp Ser His Ser Phe Ser Leu Asn 835
840 845 Ile Asp Thr Gly Ser Ile Asp Phe
Asn Glu Ser Val Gly Ile Trp Val 850 855
860 Leu Phe Lys Ile Ser Thr Pro Glu Gly Tyr Ala Lys Phe
Gly Asn Leu865 870 875
880 Glu Val Ile Glu Asn Gly Pro Val Ile Gly Glu Ala Leu Ala Arg Val
885 890 895 Lys Arg Gln Glu
Thr Lys Trp Arg Asn Gln Leu Thr Gln Leu Arg Thr 900
905 910 Glu Thr Gln Ala Ile Tyr Thr Arg Ala
Lys Gln Ala Leu Asp Asn Leu 915 920
925 Phe Ala Asn Ala Gln Asp Ser His Leu Lys Ile Gly Thr Thr
Phe Ala 930 935 940
Ala Ile Val Ala Ala Arg Lys Ile Val Gln Ser Ile Arg Glu Ala Tyr945
950 955 960 Met Ser Trp Leu Ser
Val Val Pro Gly Val Asn Tyr Pro Ile Phe Thr 965
970 975 Glu Leu Thr Glu Arg Val Gln Gln Ala Phe
Gln Leu Tyr Asp Val Arg 980 985
990 Asn Val Val Arg Asn Gly Gln Phe Leu Ser Gly Leu Ser Asp Trp
Ile 995 1000 1005 Val
Thr Pro Asp Val Lys Val Gln Glu Asp Asn Gly Asn Asn Val Leu 1010
1015 1020 Val Leu Ser Asn Trp Asp
Ala Gln Val Leu Gln Cys Leu Lys Leu Tyr1025 1030
1035 1040 Gln Asp Arg Gly Tyr Ile Leu Arg Val Thr Ala
Arg Lys Glu Gly Leu 1045 1050
1055 Gly Glu Gly Tyr Val Thr Ile Thr Asp Glu Glu Gly Asn Thr Asp Gln
1060 1065 1070 Leu Thr Phe
Gly Ala Cys Glu Glu Ile Asp Ala Ser Asn Ala Phe Ile 1075
1080 1085 Ser Thr Gly Tyr Ile Thr Lys Glu
Leu Glu Phe Phe Pro Asp Thr Glu 1090 1095
1100 Lys Val Arg Ile Glu Ile Gly Glu Thr Glu Gly Thr Phe
Gln Val Glu1105 1110 1115
1120 Ser Val Glu Leu Phe Leu Met Glu Asp Leu Cys 1125
1130 241002DNABacillus thuringiensisCDS(1)...(1002) 24atg
gtg aga gtc tat cca gac ttt gat gag atg ata agg gaa gct gct 48Met
Val Arg Val Tyr Pro Asp Phe Asp Glu Met Ile Arg Glu Ala Ala1
5 10 15cga aaa tgg tca gaa gca aat
gga ttg cta ttt caa aag gtg tcg tat 96Arg Lys Trp Ser Glu Ala Asn
Gly Leu Leu Phe Gln Lys Val Ser Tyr 20 25
30gcg gat cct tta acc aat gcg gat aca atc agt cta agt gtc
aaa ttc 144Ala Asp Pro Leu Thr Asn Ala Asp Thr Ile Ser Leu Ser Val
Lys Phe 35 40 45aaa gat atc gga
tgc cta gaa gaa tgc gtc gaa gta gaa aaa ata agc 192Lys Asp Ile Gly
Cys Leu Glu Glu Cys Val Glu Val Glu Lys Ile Ser 50 55
60att cca caa aat ttt acg aat aat acg gat caa aaa caa
aaa gaa aca 240Ile Pro Gln Asn Phe Thr Asn Asn Thr Asp Gln Lys Gln
Lys Glu Thr65 70 75
80ttg gaa act ata aca tat gta gaa aat ctg ttg act tgg gaa aat gac
288Leu Glu Thr Ile Thr Tyr Val Glu Asn Leu Leu Thr Trp Glu Asn Asp
85 90 95ttt cat ttt gtg ctt cca
gga caa aac ttt ctt acg ata cca cgt gtt 336Phe His Phe Val Leu Pro
Gly Gln Asn Phe Leu Thr Ile Pro Arg Val 100
105 110cct cgt tca gtt cat atg gat att aat cca ggt ttc
ctt gtg aat ttt 384Pro Arg Ser Val His Met Asp Ile Asn Pro Gly Phe
Leu Val Asn Phe 115 120 125ttt ggt
aga aat caa cta ttc cat acc aaa ata aga gaa cca cgt ccc 432Phe Gly
Arg Asn Gln Leu Phe His Thr Lys Ile Arg Glu Pro Arg Pro 130
135 140att cgg gcg gaa gtg ttt tta gaa cca tct agc
agt gca tcg att caa 480Ile Arg Ala Glu Val Phe Leu Glu Pro Ser Ser
Ser Ala Ser Ile Gln145 150 155
160ctg caa gtg gaa aaa caa cat gtt tct caa ccg tat caa atg gaa tta
528Leu Gln Val Glu Lys Gln His Val Ser Gln Pro Tyr Gln Met Glu Leu
165 170 175tca atg cta gga agt
att att gtg acg gca caa gat aga gga cag gaa 576Ser Met Leu Gly Ser
Ile Ile Val Thr Ala Gln Asp Arg Gly Gln Glu 180
185 190caa ggt acg gat cgc tat gtt gag tta aca gat ctc
ata cca ttc ctc 624Gln Gly Thr Asp Arg Tyr Val Glu Leu Thr Asp Leu
Ile Pro Phe Leu 195 200 205tgc ccg
cat aaa aac ttt tct tct aaa ggg cgg gca ttg ata ttc ctt 672Cys Pro
His Lys Asn Phe Ser Ser Lys Gly Arg Ala Leu Ile Phe Leu 210
215 220gaa cag gga acg ttc aag gga ata ttg agt cga
aag ata cgt gca tat 720Glu Gln Gly Thr Phe Lys Gly Ile Leu Ser Arg
Lys Ile Arg Ala Tyr225 230 235
240gcc aca caa atg ctt cat tgc gac gga aaa aca cta gaa tat gaa att
768Ala Thr Gln Met Leu His Cys Asp Gly Lys Thr Leu Glu Tyr Glu Ile
245 250 255cct tta aat aat cca
tta cct gaa tct gcc cta cga cct aaa cct atg 816Pro Leu Asn Asn Pro
Leu Pro Glu Ser Ala Leu Arg Pro Lys Pro Met 260
265 270acg att aac gct aca tca tgt gga tgt tct tct gat
aga cca tca gtc 864Thr Ile Asn Ala Thr Ser Cys Gly Cys Ser Ser Asp
Arg Pro Ser Val 275 280 285gta tct
act tcc tct cat cca tcg aat cct aca acc tat tcc cag caa 912Val Ser
Thr Ser Ser His Pro Ser Asn Pro Thr Thr Tyr Ser Gln Gln 290
295 300cct aaa cct atg acg act aac gct aca tca tgt
gga tgt tct gcc tgt 960Pro Lys Pro Met Thr Thr Asn Ala Thr Ser Cys
Gly Cys Ser Ala Cys305 310 315
320atg tct gca aca tca aat aaa aat cta tat aca gaa caa taa
1002Met Ser Ala Thr Ser Asn Lys Asn Leu Tyr Thr Glu Gln
325 33025333PRTBacillus thuringiensis 25Met Val Arg Val
Tyr Pro Asp Phe Asp Glu Met Ile Arg Glu Ala Ala1 5
10 15 Arg Lys Trp Ser Glu Ala Asn Gly Leu
Leu Phe Gln Lys Val Ser Tyr 20 25
30 Ala Asp Pro Leu Thr Asn Ala Asp Thr Ile Ser Leu Ser Val
Lys Phe 35 40 45
Lys Asp Ile Gly Cys Leu Glu Glu Cys Val Glu Val Glu Lys Ile Ser 50
55 60 Ile Pro Gln Asn Phe
Thr Asn Asn Thr Asp Gln Lys Gln Lys Glu Thr65 70
75 80 Leu Glu Thr Ile Thr Tyr Val Glu Asn Leu
Leu Thr Trp Glu Asn Asp 85 90
95 Phe His Phe Val Leu Pro Gly Gln Asn Phe Leu Thr Ile Pro Arg
Val 100 105 110 Pro
Arg Ser Val His Met Asp Ile Asn Pro Gly Phe Leu Val Asn Phe 115
120 125 Phe Gly Arg Asn Gln Leu
Phe His Thr Lys Ile Arg Glu Pro Arg Pro 130 135
140 Ile Arg Ala Glu Val Phe Leu Glu Pro Ser Ser
Ser Ala Ser Ile Gln145 150 155
160 Leu Gln Val Glu Lys Gln His Val Ser Gln Pro Tyr Gln Met Glu Leu
165 170 175 Ser Met Leu
Gly Ser Ile Ile Val Thr Ala Gln Asp Arg Gly Gln Glu 180
185 190 Gln Gly Thr Asp Arg Tyr Val Glu
Leu Thr Asp Leu Ile Pro Phe Leu 195 200
205 Cys Pro His Lys Asn Phe Ser Ser Lys Gly Arg Ala Leu
Ile Phe Leu 210 215 220
Glu Gln Gly Thr Phe Lys Gly Ile Leu Ser Arg Lys Ile Arg Ala Tyr225
230 235 240 Ala Thr Gln Met Leu
His Cys Asp Gly Lys Thr Leu Glu Tyr Glu Ile 245
250 255 Pro Leu Asn Asn Pro Leu Pro Glu Ser Ala
Leu Arg Pro Lys Pro Met 260 265
270 Thr Ile Asn Ala Thr Ser Cys Gly Cys Ser Ser Asp Arg Pro Ser
Val 275 280 285 Val
Ser Thr Ser Ser His Pro Ser Asn Pro Thr Thr Tyr Ser Gln Gln 290
295 300 Pro Lys Pro Met Thr Thr
Asn Ala Thr Ser Cys Gly Cys Ser Ala Cys305 310
315 320 Met Ser Ala Thr Ser Asn Lys Asn Leu Tyr Thr
Glu Gln 325 330
261026DNABacillus thuringiensisCDS(1)...(1026) 26atg aaa aag tct tgt aat
cca aat gaa gtg aat cca agt gca tat act 48Met Lys Lys Ser Cys Asn
Pro Asn Glu Val Asn Pro Ser Ala Tyr Thr1 5
10 15tta tac aac ttt gat gag tat gta att ggc aat ttt
ttc aat gca gtg 96Leu Tyr Asn Phe Asp Glu Tyr Val Ile Gly Asn Phe
Phe Asn Ala Val 20 25 30caa
cac cct ata aat ctt tcc ttc aga gaa gat gaa tgg aat ttt gat 144Gln
His Pro Ile Asn Leu Ser Phe Arg Glu Asp Glu Trp Asn Phe Asp 35
40 45tca gac tat att cct acc cca act tct
gac caa cta act agc cct tgt 192Ser Asp Tyr Ile Pro Thr Pro Thr Ser
Asp Gln Leu Thr Ser Pro Cys 50 55
60aaa tct gtt cca acc agc gca ccg tgt aaa tat ggt cca atc gat cct
240Lys Ser Val Pro Thr Ser Ala Pro Cys Lys Tyr Gly Pro Ile Asp Pro65
70 75 80tat gag tgg ata gaa
tgg cta gac ctc gta gca gaa ggg aca gat tta 288Tyr Glu Trp Ile Glu
Trp Leu Asp Leu Val Ala Glu Gly Thr Asp Leu 85
90 95tat gaa gaa atc caa aat gca aat aca aaa acc
ccg ttt att aat gat 336Tyr Glu Glu Ile Gln Asn Ala Asn Thr Lys Thr
Pro Phe Ile Asn Asp 100 105
110gaa tat tac ttt aat aat aca tca tct gaa aca caa cca tat cag acc
384Glu Tyr Tyr Phe Asn Asn Thr Ser Ser Glu Thr Gln Pro Tyr Gln Thr
115 120 125att tct cat tca gaa aca ttg
acc aca acc acg aca aac aca aca aca 432Ile Ser His Ser Glu Thr Leu
Thr Thr Thr Thr Thr Asn Thr Thr Thr 130 135
140caa gga tgt aaa ata aat cca aaa gtc agt tat tcg aga aaa acg aaa
480Gln Gly Cys Lys Ile Asn Pro Lys Val Ser Tyr Ser Arg Lys Thr Lys145
150 155 160gtt aaa gtc aaa
att gtt gat gta gaa aag gga ttt aac acg gaa ata 528Val Lys Val Lys
Ile Val Asp Val Glu Lys Gly Phe Asn Thr Glu Ile 165
170 175ggg gca gaa tat aac ttt agc gat aca aac
aca tac aca gaa aca gca 576Gly Ala Glu Tyr Asn Phe Ser Asp Thr Asn
Thr Tyr Thr Glu Thr Ala 180 185
190act cgg act gtg aca gtt cca tca atg acg aca tat gtt cca ccc tat
624Thr Arg Thr Val Thr Val Pro Ser Met Thr Thr Tyr Val Pro Pro Tyr
195 200 205acc tct gcg tat gtg acg gtc
gta tta gaa aga gga tat tat gaa gct 672Thr Ser Ala Tyr Val Thr Val
Val Leu Glu Arg Gly Tyr Tyr Glu Ala 210 215
220tat aat att cca atc gac acg aat tta tat gga aga ttt gaa ctc atc
720Tyr Asn Ile Pro Ile Asp Thr Asn Leu Tyr Gly Arg Phe Glu Leu Ile225
230 235 240tac acc aac ctc
gat ggc agc aac cct cga tcg gcg ggt gtt ctg gat 768Tyr Thr Asn Leu
Asp Gly Ser Asn Pro Arg Ser Ala Gly Val Leu Asp 245
250 255cta tat cct ttc gta gaa tta att aca aca
tgt tgt caa aat tgt agt 816Leu Tyr Pro Phe Val Glu Leu Ile Thr Thr
Cys Cys Gln Asn Cys Ser 260 265
270cag tgt gtg cca gac atg att cag cct gat cgt gac aat caa aca gtt
864Gln Cys Val Pro Asp Met Ile Gln Pro Asp Arg Asp Asn Gln Thr Val
275 280 285cgt ttt act gga aga ggg gat
ctt ata tcg gat ttt gca gcg aat act 912Arg Phe Thr Gly Arg Gly Asp
Leu Ile Ser Asp Phe Ala Ala Asn Thr 290 295
300tta acc gta acg act aca ttt gtt gac aat gca aca gga gca acc gtt
960Leu Thr Val Thr Thr Thr Phe Val Asp Asn Ala Thr Gly Ala Thr Val305
310 315 320tcg caa cac gta
gaa tcc gta cca gtt cag tat gga ccc gct acc aca 1008Ser Gln His Val
Glu Ser Val Pro Val Gln Tyr Gly Pro Ala Thr Thr 325
330 335gta gta aac aca tct aaa
1026Val Val Asn Thr Ser Lys
34027342PRTBacillus thuringiensis 27Met Lys Lys Ser Cys Asn Pro Asn Glu
Val Asn Pro Ser Ala Tyr Thr1 5 10
15 Leu Tyr Asn Phe Asp Glu Tyr Val Ile Gly Asn Phe Phe Asn
Ala Val 20 25 30
Gln His Pro Ile Asn Leu Ser Phe Arg Glu Asp Glu Trp Asn Phe Asp 35
40 45 Ser Asp Tyr Ile Pro
Thr Pro Thr Ser Asp Gln Leu Thr Ser Pro Cys 50 55
60 Lys Ser Val Pro Thr Ser Ala Pro Cys Lys
Tyr Gly Pro Ile Asp Pro65 70 75
80 Tyr Glu Trp Ile Glu Trp Leu Asp Leu Val Ala Glu Gly Thr Asp
Leu 85 90 95 Tyr
Glu Glu Ile Gln Asn Ala Asn Thr Lys Thr Pro Phe Ile Asn Asp
100 105 110 Glu Tyr Tyr Phe Asn
Asn Thr Ser Ser Glu Thr Gln Pro Tyr Gln Thr 115
120 125 Ile Ser His Ser Glu Thr Leu Thr Thr
Thr Thr Thr Asn Thr Thr Thr 130 135
140 Gln Gly Cys Lys Ile Asn Pro Lys Val Ser Tyr Ser Arg
Lys Thr Lys145 150 155
160 Val Lys Val Lys Ile Val Asp Val Glu Lys Gly Phe Asn Thr Glu Ile
165 170 175 Gly Ala Glu Tyr
Asn Phe Ser Asp Thr Asn Thr Tyr Thr Glu Thr Ala 180
185 190 Thr Arg Thr Val Thr Val Pro Ser Met
Thr Thr Tyr Val Pro Pro Tyr 195 200
205 Thr Ser Ala Tyr Val Thr Val Val Leu Glu Arg Gly Tyr Tyr
Glu Ala 210 215 220
Tyr Asn Ile Pro Ile Asp Thr Asn Leu Tyr Gly Arg Phe Glu Leu Ile225
230 235 240 Tyr Thr Asn Leu Asp
Gly Ser Asn Pro Arg Ser Ala Gly Val Leu Asp 245
250 255 Leu Tyr Pro Phe Val Glu Leu Ile Thr Thr
Cys Cys Gln Asn Cys Ser 260 265
270 Gln Cys Val Pro Asp Met Ile Gln Pro Asp Arg Asp Asn Gln Thr
Val 275 280 285 Arg
Phe Thr Gly Arg Gly Asp Leu Ile Ser Asp Phe Ala Ala Asn Thr 290
295 300 Leu Thr Val Thr Thr Thr
Phe Val Asp Asn Ala Thr Gly Ala Thr Val305 310
315 320 Ser Gln His Val Glu Ser Val Pro Val Gln Tyr
Gly Pro Ala Thr Thr 325 330
335 Val Val Asn Thr Ser Lys 340 283009DNABacillus
thuringiensisCDS(1)...(3009) 28atg aaa agg atg aag aaa aag cta gca agt
gtt gta acc tgt aca tta 48Met Lys Arg Met Lys Lys Lys Leu Ala Ser
Val Val Thr Cys Thr Leu1 5 10
15tta gct cct atg ttt ttg aat ggg aac gta gac act gta ttt gca gat
96Leu Ala Pro Met Phe Leu Asn Gly Asn Val Asp Thr Val Phe Ala Asp
20 25 30agt aaa aca aat caa att
tct tca aca cag gaa aac caa aag aat gag 144Ser Lys Thr Asn Gln Ile
Ser Ser Thr Gln Glu Asn Gln Lys Asn Glu 35 40
45atg gat cga aaa gga cta ctt ggt tat tat ttt aaa gga aaa
gat ttt 192Met Asp Arg Lys Gly Leu Leu Gly Tyr Tyr Phe Lys Gly Lys
Asp Phe 50 55 60aat aat ctt act ata
ttt gct cca aca cgt gag aat act ctt att tat 240Asn Asn Leu Thr Ile
Phe Ala Pro Thr Arg Glu Asn Thr Leu Ile Tyr65 70
75 80gat tta gaa aca gcg aat tct tta tta gat
aag caa caa caa acc tat 288Asp Leu Glu Thr Ala Asn Ser Leu Leu Asp
Lys Gln Gln Gln Thr Tyr 85 90
95caa tct att cgt tgg atc ggt tta ata aaa agc aaa aaa gct gga gat
336Gln Ser Ile Arg Trp Ile Gly Leu Ile Lys Ser Lys Lys Ala Gly Asp
100 105 110ttt acc ttt caa tta tcg
gat gat gag cat gct att ata gaa atc gat 384Phe Thr Phe Gln Leu Ser
Asp Asp Glu His Ala Ile Ile Glu Ile Asp 115 120
125ggg aaa gtt att tcg caa aaa ggc caa aag aaa caa gtt gtt
cat tta 432Gly Lys Val Ile Ser Gln Lys Gly Gln Lys Lys Gln Val Val
His Leu 130 135 140gaa aaa gat aaa tta
gtt ccc atc aaa att gaa tat caa tct gat aaa 480Glu Lys Asp Lys Leu
Val Pro Ile Lys Ile Glu Tyr Gln Ser Asp Lys145 150
155 160gcg tta aac cca gat agt caa atg ttt aaa
gaa ttg aaa tta ttt aaa 528Ala Leu Asn Pro Asp Ser Gln Met Phe Lys
Glu Leu Lys Leu Phe Lys 165 170
175ata aat agt caa aaa caa tct cag caa gtg caa caa gac gaa ttg aga
576Ile Asn Ser Gln Lys Gln Ser Gln Gln Val Gln Gln Asp Glu Leu Arg
180 185 190aat cct gaa ttt ggt aaa
gaa aaa act caa aca tat tta aag aaa gca 624Asn Pro Glu Phe Gly Lys
Glu Lys Thr Gln Thr Tyr Leu Lys Lys Ala 195 200
205tcg aaa agc agc ttg ttt agc aat aaa agt aaa cga gat ata
gat gaa 672Ser Lys Ser Ser Leu Phe Ser Asn Lys Ser Lys Arg Asp Ile
Asp Glu 210 215 220gat ata gat gag gat
aca gat aca gat gga gat gcc att cct gat gta 720Asp Ile Asp Glu Asp
Thr Asp Thr Asp Gly Asp Ala Ile Pro Asp Val225 230
235 240tgg gaa gaa aat ggg tat acc atc aaa gga
aga gta gct gtt aaa tgg 768Trp Glu Glu Asn Gly Tyr Thr Ile Lys Gly
Arg Val Ala Val Lys Trp 245 250
255gac gaa gga tta gct gat aag gga tat aaa aag ttt gtt tcc aat cct
816Asp Glu Gly Leu Ala Asp Lys Gly Tyr Lys Lys Phe Val Ser Asn Pro
260 265 270ttt aga cag cac act gct
ggt gac ccc tat agt gac tat gaa aag gca 864Phe Arg Gln His Thr Ala
Gly Asp Pro Tyr Ser Asp Tyr Glu Lys Ala 275 280
285tca aaa gat ttg gat tta tct aat gca aaa gaa aca ttt aat
cca ttg 912Ser Lys Asp Leu Asp Leu Ser Asn Ala Lys Glu Thr Phe Asn
Pro Leu 290 295 300gtg gct gct ttt cca
agt gtc aat gtt agc ttg gaa aat gtc acc ata 960Val Ala Ala Phe Pro
Ser Val Asn Val Ser Leu Glu Asn Val Thr Ile305 310
315 320tca aaa gat gaa aat aaa act gct gaa att
gcg tct act tca tcg aat 1008Ser Lys Asp Glu Asn Lys Thr Ala Glu Ile
Ala Ser Thr Ser Ser Asn 325 330
335aat tgg tcc tat aca aat aca gag ggg gca tct att gaa gct gga att
1056Asn Trp Ser Tyr Thr Asn Thr Glu Gly Ala Ser Ile Glu Ala Gly Ile
340 345 350gga cca gaa ggt ttg ttg
tct ttt gga gta agt gcc aat tat caa cat 1104Gly Pro Glu Gly Leu Leu
Ser Phe Gly Val Ser Ala Asn Tyr Gln His 355 360
365tct gaa aca gtg gcc aaa gag tgg ggt aca act aag gga gac
gca aca 1152Ser Glu Thr Val Ala Lys Glu Trp Gly Thr Thr Lys Gly Asp
Ala Thr 370 375 380caa tat aat aca gct
tca gca gga tat cta aat gcc aat gtt cga tat 1200Gln Tyr Asn Thr Ala
Ser Ala Gly Tyr Leu Asn Ala Asn Val Arg Tyr385 390
395 400aat aat gta ggg acg gca gcc att tat gat
gtg aaa cct aca acg aat 1248Asn Asn Val Gly Thr Ala Ala Ile Tyr Asp
Val Lys Pro Thr Thr Asn 405 410
415ttt gta tta gat aag act aca ctc gcg acg att aag gca aaa gaa aat
1296Phe Val Leu Asp Lys Thr Thr Leu Ala Thr Ile Lys Ala Lys Glu Asn
420 425 430gcc acg gct gat cat ata
ata cca ggg aat agt tac ccg gaa aaa ggg 1344Ala Thr Ala Asp His Ile
Ile Pro Gly Asn Ser Tyr Pro Glu Lys Gly 435 440
445aaa aat gga att gcg ata act act atg gat gat ttt aac tct
cat cct 1392Lys Asn Gly Ile Ala Ile Thr Thr Met Asp Asp Phe Asn Ser
His Pro 450 455 460att act cta aat aaa
caa caa cta gat aag ttg tta tat aat gtt aca 1440Ile Thr Leu Asn Lys
Gln Gln Leu Asp Lys Leu Leu Tyr Asn Val Thr465 470
475 480cca ctt atg ttg gaa act acc caa gtt gag
ggt acg tat aag aaa aaa 1488Pro Leu Met Leu Glu Thr Thr Gln Val Glu
Gly Thr Tyr Lys Lys Lys 485 490
495gat gta gat ggt aat atc att act gga ggg aca tgg agt gga gtg aca
1536Asp Val Asp Gly Asn Ile Ile Thr Gly Gly Thr Trp Ser Gly Val Thr
500 505 510caa caa att gag gcg caa
act gct tct att att gtt gat act gga gag 1584Gln Gln Ile Glu Ala Gln
Thr Ala Ser Ile Ile Val Asp Thr Gly Glu 515 520
525ggt gtt tcg gaa aaa cgt att gca gca aaa gat tat gat gac
ccc gaa 1632Gly Val Ser Glu Lys Arg Ile Ala Ala Lys Asp Tyr Asp Asp
Pro Glu 530 535 540gat aaa aca ccg tct
tta act cta aaa gat gcc ctg aaa att gga tat 1680Asp Lys Thr Pro Ser
Leu Thr Leu Lys Asp Ala Leu Lys Ile Gly Tyr545 550
555 560cca gaa gaa att gaa gaa aaa aat gat tta
tta tat tat aaa gga aaa 1728Pro Glu Glu Ile Glu Glu Lys Asn Asp Leu
Leu Tyr Tyr Lys Gly Lys 565 570
575ata ata tct gaa tca agt gtg atg act ttt ctt gat aat gga acc tcc
1776Ile Ile Ser Glu Ser Ser Val Met Thr Phe Leu Asp Asn Gly Thr Ser
580 585 590gaa aaa gtt aaa aaa caa
atc gag gat aaa act gga aaa ttt aaa gac 1824Glu Lys Val Lys Lys Gln
Ile Glu Asp Lys Thr Gly Lys Phe Lys Asp 595 600
605gta caa cat ttg tat gat gtg aaa cta aca cct gga atg aat
ttt act 1872Val Gln His Leu Tyr Asp Val Lys Leu Thr Pro Gly Met Asn
Phe Thr 610 615 620att aaa tta gct tca
ata tac gat agt gtc gat aat ttt agt ggc agt 1920Ile Lys Leu Ala Ser
Ile Tyr Asp Ser Val Asp Asn Phe Ser Gly Ser625 630
635 640caa tca tta ggg gca tta aat agt ata agt
aag gtt gct gga gga aat 1968Gln Ser Leu Gly Ala Leu Asn Ser Ile Ser
Lys Val Ala Gly Gly Asn 645 650
655aca ggg aaa aat caa tat caa tca tca tct tct aat gcg tat att tct
2016Thr Gly Lys Asn Gln Tyr Gln Ser Ser Ser Ser Asn Ala Tyr Ile Ser
660 665 670tta tct tcg agt aca aaa
gga gag ttg aat aaa aat act acg tac tat 2064Leu Ser Ser Ser Thr Lys
Gly Glu Leu Asn Lys Asn Thr Thr Tyr Tyr 675 680
685ctt agc atg tat atg aga gca gat gct gat aca gaa cct acc
ata gaa 2112Leu Ser Met Tyr Met Arg Ala Asp Ala Asp Thr Glu Pro Thr
Ile Glu 690 695 700cta aaa gga gaa aaa
tcc aca ata aaa agc caa aaa gtt aaa cta aac 2160Leu Lys Gly Glu Lys
Ser Thr Ile Lys Ser Gln Lys Val Lys Leu Asn705 710
715 720aat aag gga tat cag agg gta gat att tta
gta gaa aat act gag tcg 2208Asn Lys Gly Tyr Gln Arg Val Asp Ile Leu
Val Glu Asn Thr Glu Ser 725 730
735aat cca att cat caa att tat gta cac ggt aac aat aag aca aat gtc
2256Asn Pro Ile His Gln Ile Tyr Val His Gly Asn Asn Lys Thr Asn Val
740 745 750tat tgg gac gat gtg tct
ctt aca gag gta tct gct ata aaa caa gaa 2304Tyr Trp Asp Asp Val Ser
Leu Thr Glu Val Ser Ala Ile Lys Gln Glu 755 760
765tta ccc gat ata tca gat aaa gag ata caa agg gct cat aca
ttt aag 2352Leu Pro Asp Ile Ser Asp Lys Glu Ile Gln Arg Ala His Thr
Phe Lys 770 775 780aaa gaa cag tta agt
tta gat ggt aag tat atg aat gaa ttg aca tta 2400Lys Glu Gln Leu Ser
Leu Asp Gly Lys Tyr Met Asn Glu Leu Thr Leu785 790
795 800cac gtt gat tcg tta aaa gac aag aac aat
aag ccg gtt cag ttc agt 2448His Val Asp Ser Leu Lys Asp Lys Asn Asn
Lys Pro Val Gln Phe Ser 805 810
815tat aaa gtg aaa gat ggt gaa aaa gat tta gga act aaa tcc tat aca
2496Tyr Lys Val Lys Asp Gly Glu Lys Asp Leu Gly Thr Lys Ser Tyr Thr
820 825 830cct gat aag cag gga aat
ata aac att aac ttt cta gat tac aat cgt 2544Pro Asp Lys Gln Gly Asn
Ile Asn Ile Asn Phe Leu Asp Tyr Asn Arg 835 840
845gga ttt gga att tct aag gat cat aaa att caa att tat gcg
gta cgt 2592Gly Phe Gly Ile Ser Lys Asp His Lys Ile Gln Ile Tyr Ala
Val Arg 850 855 860aaa gac caa gag gtg
aaa gta gca gaa cta aaa aac tat aat atg agt 2640Lys Asp Gln Glu Val
Lys Val Ala Glu Leu Lys Asn Tyr Asn Met Ser865 870
875 880gga act atc aga ttt agc aat gat gga gaa
agt ggc ctt cca gag ata 2688Gly Thr Ile Arg Phe Ser Asn Asp Gly Glu
Ser Gly Leu Pro Glu Ile 885 890
895tat gga tat att ttc atg act cca gaa ggt caa tat cct gtt tct cct
2736Tyr Gly Tyr Ile Phe Met Thr Pro Glu Gly Gln Tyr Pro Val Ser Pro
900 905 910gta gga ggt ata cac caa
ata tgg tca aga tat tat aca agc act tac 2784Val Gly Gly Ile His Gln
Ile Trp Ser Arg Tyr Tyr Thr Ser Thr Tyr 915 920
925aag tgg agt act caa tat agt tat gat ttt gca tcc ttt aat
agt gat 2832Lys Trp Ser Thr Gln Tyr Ser Tyr Asp Phe Ala Ser Phe Asn
Ser Asp 930 935 940ata aaa aca gtt cat
ttt aat gga tat gta aag gaa ctg gat gat acg 2880Ile Lys Thr Val His
Phe Asn Gly Tyr Val Lys Glu Leu Asp Asp Thr945 950
955 960aat ggc gac gac ata ctg gct tat ctt gaa
aat aaa tat gaa tct cat 2928Asn Gly Asp Asp Ile Leu Ala Tyr Leu Glu
Asn Lys Tyr Glu Ser His 965 970
975ggt tta gag ggg agt gta gtg tta gaa gga gat gaa agg ggc agt aat
2976Gly Leu Glu Gly Ser Val Val Leu Glu Gly Asp Glu Arg Gly Ser Asn
980 985 990gtg act gtt gaa tat cat
ata aaa ttg aaa taa 3009Val Thr Val Glu Tyr His
Ile Lys Leu Lys 995 1000291002PRTBacillus
thuringiensis 29Met Lys Arg Met Lys Lys Lys Leu Ala Ser Val Val Thr Cys
Thr Leu1 5 10 15
Leu Ala Pro Met Phe Leu Asn Gly Asn Val Asp Thr Val Phe Ala Asp
20 25 30 Ser Lys Thr Asn Gln
Ile Ser Ser Thr Gln Glu Asn Gln Lys Asn Glu 35 40
45 Met Asp Arg Lys Gly Leu Leu Gly Tyr Tyr
Phe Lys Gly Lys Asp Phe 50 55 60
Asn Asn Leu Thr Ile Phe Ala Pro Thr Arg Glu Asn Thr Leu Ile
Tyr65 70 75 80 Asp
Leu Glu Thr Ala Asn Ser Leu Leu Asp Lys Gln Gln Gln Thr Tyr
85 90 95 Gln Ser Ile Arg Trp Ile
Gly Leu Ile Lys Ser Lys Lys Ala Gly Asp 100
105 110 Phe Thr Phe Gln Leu Ser Asp Asp Glu His
Ala Ile Ile Glu Ile Asp 115 120
125 Gly Lys Val Ile Ser Gln Lys Gly Gln Lys Lys Gln Val Val
His Leu 130 135 140
Glu Lys Asp Lys Leu Val Pro Ile Lys Ile Glu Tyr Gln Ser Asp Lys145
150 155 160 Ala Leu Asn Pro Asp
Ser Gln Met Phe Lys Glu Leu Lys Leu Phe Lys 165
170 175 Ile Asn Ser Gln Lys Gln Ser Gln Gln Val
Gln Gln Asp Glu Leu Arg 180 185
190 Asn Pro Glu Phe Gly Lys Glu Lys Thr Gln Thr Tyr Leu Lys Lys
Ala 195 200 205 Ser
Lys Ser Ser Leu Phe Ser Asn Lys Ser Lys Arg Asp Ile Asp Glu 210
215 220 Asp Ile Asp Glu Asp Thr
Asp Thr Asp Gly Asp Ala Ile Pro Asp Val225 230
235 240 Trp Glu Glu Asn Gly Tyr Thr Ile Lys Gly Arg
Val Ala Val Lys Trp 245 250
255 Asp Glu Gly Leu Ala Asp Lys Gly Tyr Lys Lys Phe Val Ser Asn Pro
260 265 270 Phe Arg Gln
His Thr Ala Gly Asp Pro Tyr Ser Asp Tyr Glu Lys Ala 275
280 285 Ser Lys Asp Leu Asp Leu Ser Asn
Ala Lys Glu Thr Phe Asn Pro Leu 290 295
300 Val Ala Ala Phe Pro Ser Val Asn Val Ser Leu Glu Asn
Val Thr Ile305 310 315
320 Ser Lys Asp Glu Asn Lys Thr Ala Glu Ile Ala Ser Thr Ser Ser Asn
325 330 335 Asn Trp Ser Tyr
Thr Asn Thr Glu Gly Ala Ser Ile Glu Ala Gly Ile 340
345 350 Gly Pro Glu Gly Leu Leu Ser Phe Gly
Val Ser Ala Asn Tyr Gln His 355 360
365 Ser Glu Thr Val Ala Lys Glu Trp Gly Thr Thr Lys Gly Asp
Ala Thr 370 375 380
Gln Tyr Asn Thr Ala Ser Ala Gly Tyr Leu Asn Ala Asn Val Arg Tyr385
390 395 400 Asn Asn Val Gly Thr
Ala Ala Ile Tyr Asp Val Lys Pro Thr Thr Asn 405
410 415 Phe Val Leu Asp Lys Thr Thr Leu Ala Thr
Ile Lys Ala Lys Glu Asn 420 425
430 Ala Thr Ala Asp His Ile Ile Pro Gly Asn Ser Tyr Pro Glu Lys
Gly 435 440 445 Lys
Asn Gly Ile Ala Ile Thr Thr Met Asp Asp Phe Asn Ser His Pro 450
455 460 Ile Thr Leu Asn Lys Gln
Gln Leu Asp Lys Leu Leu Tyr Asn Val Thr465 470
475 480 Pro Leu Met Leu Glu Thr Thr Gln Val Glu Gly
Thr Tyr Lys Lys Lys 485 490
495 Asp Val Asp Gly Asn Ile Ile Thr Gly Gly Thr Trp Ser Gly Val Thr
500 505 510 Gln Gln Ile
Glu Ala Gln Thr Ala Ser Ile Ile Val Asp Thr Gly Glu 515
520 525 Gly Val Ser Glu Lys Arg Ile Ala
Ala Lys Asp Tyr Asp Asp Pro Glu 530 535
540 Asp Lys Thr Pro Ser Leu Thr Leu Lys Asp Ala Leu Lys
Ile Gly Tyr545 550 555
560 Pro Glu Glu Ile Glu Glu Lys Asn Asp Leu Leu Tyr Tyr Lys Gly Lys
565 570 575 Ile Ile Ser Glu
Ser Ser Val Met Thr Phe Leu Asp Asn Gly Thr Ser 580
585 590 Glu Lys Val Lys Lys Gln Ile Glu Asp
Lys Thr Gly Lys Phe Lys Asp 595 600
605 Val Gln His Leu Tyr Asp Val Lys Leu Thr Pro Gly Met Asn
Phe Thr 610 615 620
Ile Lys Leu Ala Ser Ile Tyr Asp Ser Val Asp Asn Phe Ser Gly Ser625
630 635 640 Gln Ser Leu Gly Ala
Leu Asn Ser Ile Ser Lys Val Ala Gly Gly Asn 645
650 655 Thr Gly Lys Asn Gln Tyr Gln Ser Ser Ser
Ser Asn Ala Tyr Ile Ser 660 665
670 Leu Ser Ser Ser Thr Lys Gly Glu Leu Asn Lys Asn Thr Thr Tyr
Tyr 675 680 685 Leu
Ser Met Tyr Met Arg Ala Asp Ala Asp Thr Glu Pro Thr Ile Glu 690
695 700 Leu Lys Gly Glu Lys Ser
Thr Ile Lys Ser Gln Lys Val Lys Leu Asn705 710
715 720 Asn Lys Gly Tyr Gln Arg Val Asp Ile Leu Val
Glu Asn Thr Glu Ser 725 730
735 Asn Pro Ile His Gln Ile Tyr Val His Gly Asn Asn Lys Thr Asn Val
740 745 750 Tyr Trp Asp
Asp Val Ser Leu Thr Glu Val Ser Ala Ile Lys Gln Glu 755
760 765 Leu Pro Asp Ile Ser Asp Lys Glu
Ile Gln Arg Ala His Thr Phe Lys 770 775
780 Lys Glu Gln Leu Ser Leu Asp Gly Lys Tyr Met Asn Glu
Leu Thr Leu785 790 795
800 His Val Asp Ser Leu Lys Asp Lys Asn Asn Lys Pro Val Gln Phe Ser
805 810 815 Tyr Lys Val Lys
Asp Gly Glu Lys Asp Leu Gly Thr Lys Ser Tyr Thr 820
825 830 Pro Asp Lys Gln Gly Asn Ile Asn Ile
Asn Phe Leu Asp Tyr Asn Arg 835 840
845 Gly Phe Gly Ile Ser Lys Asp His Lys Ile Gln Ile Tyr Ala
Val Arg 850 855 860
Lys Asp Gln Glu Val Lys Val Ala Glu Leu Lys Asn Tyr Asn Met Ser865
870 875 880 Gly Thr Ile Arg Phe
Ser Asn Asp Gly Glu Ser Gly Leu Pro Glu Ile 885
890 895 Tyr Gly Tyr Ile Phe Met Thr Pro Glu Gly
Gln Tyr Pro Val Ser Pro 900 905
910 Val Gly Gly Ile His Gln Ile Trp Ser Arg Tyr Tyr Thr Ser Thr
Tyr 915 920 925 Lys
Trp Ser Thr Gln Tyr Ser Tyr Asp Phe Ala Ser Phe Asn Ser Asp 930
935 940 Ile Lys Thr Val His Phe
Asn Gly Tyr Val Lys Glu Leu Asp Asp Thr945 950
955 960 Asn Gly Asp Asp Ile Leu Ala Tyr Leu Glu Asn
Lys Tyr Glu Ser His 965 970
975 Gly Leu Glu Gly Ser Val Val Leu Glu Gly Asp Glu Arg Gly Ser Asn
980 985 990 Val Thr Val
Glu Tyr His Ile Lys Leu Lys 995 1000
301347DNABacillus thuringiensisCDS(1)...(1347) 30atg cat aaa caa aca ata
aaa aat tta tct att tgc ata gca act gta 48Met His Lys Gln Thr Ile
Lys Asn Leu Ser Ile Cys Ile Ala Thr Val1 5
10 15tca cta tta gga caa tac ttt ata tcc tct act acg
gta tac gct gct 96Ser Leu Leu Gly Gln Tyr Phe Ile Ser Ser Thr Thr
Val Tyr Ala Ala 20 25 30gaa
aat caa att aac tca tta aat cta aaa gta gag caa att cta gat 144Glu
Asn Gln Ile Asn Ser Leu Asn Leu Lys Val Glu Gln Ile Leu Asp 35
40 45ttt gga aga gat aaa gag aaa gca aaa
gaa tgg gcc gat act tac ttt 192Phe Gly Arg Asp Lys Glu Lys Ala Lys
Glu Trp Ala Asp Thr Tyr Phe 50 55
60aaa gat tgg aag aaa aca att aat aat gaa caa aaa aaa ctt tta aat
240Lys Asp Trp Lys Lys Thr Ile Asn Asn Glu Gln Lys Lys Leu Leu Asn65
70 75 80gat att aaa cgc tta
aca caa tta aat gaa aaa ata ggt aaa ttc gat 288Asp Ile Lys Arg Leu
Thr Gln Leu Asn Glu Lys Ile Gly Lys Phe Asp 85
90 95caa aat tca gag atg ttt tct aaa aaa gat aaa
gag gat ata gat aaa 336Gln Asn Ser Glu Met Phe Ser Lys Lys Asp Lys
Glu Asp Ile Asp Lys 100 105
110ata gac aaa gct ttg aat aat aaa aat gca aaa tta acc aaa tct ttg
384Ile Asp Lys Ala Leu Asn Asn Lys Asn Ala Lys Leu Thr Lys Ser Leu
115 120 125aat gtg tat aaa aat ttg aat
ggc aaa gat tta gga tat gta gaa ggt 432Asn Val Tyr Lys Asn Leu Asn
Gly Lys Asp Leu Gly Tyr Val Glu Gly 130 135
140tat ttt aat gta ccg aat tcc cca aat aaa ata gat aga aca aaa tat
480Tyr Phe Asn Val Pro Asn Ser Pro Asn Lys Ile Asp Arg Thr Lys Tyr145
150 155 160aat aaa tta gtt
aat gag ttt aag tat ggg gct atc aat aca ttc atg 528Asn Lys Leu Val
Asn Glu Phe Lys Tyr Gly Ala Ile Asn Thr Phe Met 165
170 175aat aca gac tta aca caa gat act aca aat
aaa tca aca cct att ttg 576Asn Thr Asp Leu Thr Gln Asp Thr Thr Asn
Lys Ser Thr Pro Ile Leu 180 185
190ctt tca ttg aag cta cca aaa gga aca aaa ata gga caa tta aat gaa
624Leu Ser Leu Lys Leu Pro Lys Gly Thr Lys Ile Gly Gln Leu Asn Glu
195 200 205gaa cat ata ata aca gac aga
aac tta gga att gaa ata aaa aaa aca 672Glu His Ile Ile Thr Asp Arg
Asn Leu Gly Ile Glu Ile Lys Lys Thr 210 215
220agt att att gtt gaa aaa gga aga gaa gtt att aaa cta gaa gga gac
720Ser Ile Ile Val Glu Lys Gly Arg Glu Val Ile Lys Leu Glu Gly Asp225
230 235 240gta gta cca aaa
act aaa att caa gaa aaa gta aaa aaa gca gaa agt 768Val Val Pro Lys
Thr Lys Ile Gln Glu Lys Val Lys Lys Ala Glu Ser 245
250 255gat ttg aat caa aaa ttt aaa gaa ata acg
ggt tta aaa caa aac tta 816Asp Leu Asn Gln Lys Phe Lys Glu Ile Thr
Gly Leu Lys Gln Asn Leu 260 265
270cta agt ctg aaa ata gat aat cta tat aca tca gct agc att gac aga
864Leu Ser Leu Lys Ile Asp Asn Leu Tyr Thr Ser Ala Ser Ile Asp Arg
275 280 285acg gaa aca gtt ata aaa caa
tta gtc agc aat gta cca aac aat tta 912Thr Glu Thr Val Ile Lys Gln
Leu Val Ser Asn Val Pro Asn Asn Leu 290 295
300ttg tta aat ata atg aaa aat atg aat aat aaa aca tta ttt act att
960Leu Leu Asn Ile Met Lys Asn Met Asn Asn Lys Thr Leu Phe Thr Ile305
310 315 320aca gat aaa att
cta ata ccc ggc aag gaa ggt gta tta ggt tat tat 1008Thr Asp Lys Ile
Leu Ile Pro Gly Lys Glu Gly Val Leu Gly Tyr Tyr 325
330 335gat acc att tct aaa aca tta ttt ata caa
att gat cat ttg ggg cat 1056Asp Thr Ile Ser Lys Thr Leu Phe Ile Gln
Ile Asp His Leu Gly His 340 345
350aaa aac aat gaa gga aat gac act aat act ctt ctt cat gaa ttt ggt
1104Lys Asn Asn Glu Gly Asn Asp Thr Asn Thr Leu Leu His Glu Phe Gly
355 360 365cat gct gta gat cat ttg gca
aaa ggg gag ata caa tca aaa tct agt 1152His Ala Val Asp His Leu Ala
Lys Gly Glu Ile Gln Ser Lys Ser Ser 370 375
380aag ttc att gaa ata ttt aat cga gag aga ggt aat att aca ata gaa
1200Lys Phe Ile Glu Ile Phe Asn Arg Glu Arg Gly Asn Ile Thr Ile Glu385
390 395 400cca tat att aaa
caa gat gca gcg gaa ttt ttt gca ggt gtt ttt aat 1248Pro Tyr Ile Lys
Gln Asp Ala Ala Glu Phe Phe Ala Gly Val Phe Asn 405
410 415tac tta tat tca cct aaa ata tca gat aga
gaa caa att caa aaa gaa 1296Tyr Leu Tyr Ser Pro Lys Ile Ser Asp Arg
Glu Gln Ile Gln Lys Glu 420 425
430gca cct gat gct tgt aaa ttt atc cga aat tta ata cat ggt cta cat
1344Ala Pro Asp Ala Cys Lys Phe Ile Arg Asn Leu Ile His Gly Leu His
435 440 445tga
134731448PRTBacillus thuringiensis
31Met His Lys Gln Thr Ile Lys Asn Leu Ser Ile Cys Ile Ala Thr Val1
5 10 15 Ser Leu Leu Gly
Gln Tyr Phe Ile Ser Ser Thr Thr Val Tyr Ala Ala 20
25 30 Glu Asn Gln Ile Asn Ser Leu Asn Leu
Lys Val Glu Gln Ile Leu Asp 35 40
45 Phe Gly Arg Asp Lys Glu Lys Ala Lys Glu Trp Ala Asp Thr
Tyr Phe 50 55 60
Lys Asp Trp Lys Lys Thr Ile Asn Asn Glu Gln Lys Lys Leu Leu Asn65
70 75 80 Asp Ile Lys Arg Leu
Thr Gln Leu Asn Glu Lys Ile Gly Lys Phe Asp 85
90 95 Gln Asn Ser Glu Met Phe Ser Lys Lys Asp
Lys Glu Asp Ile Asp Lys 100 105
110 Ile Asp Lys Ala Leu Asn Asn Lys Asn Ala Lys Leu Thr Lys Ser
Leu 115 120 125 Asn
Val Tyr Lys Asn Leu Asn Gly Lys Asp Leu Gly Tyr Val Glu Gly 130
135 140 Tyr Phe Asn Val Pro Asn
Ser Pro Asn Lys Ile Asp Arg Thr Lys Tyr145 150
155 160 Asn Lys Leu Val Asn Glu Phe Lys Tyr Gly Ala
Ile Asn Thr Phe Met 165 170
175 Asn Thr Asp Leu Thr Gln Asp Thr Thr Asn Lys Ser Thr Pro Ile Leu
180 185 190 Leu Ser Leu
Lys Leu Pro Lys Gly Thr Lys Ile Gly Gln Leu Asn Glu 195
200 205 Glu His Ile Ile Thr Asp Arg Asn
Leu Gly Ile Glu Ile Lys Lys Thr 210 215
220 Ser Ile Ile Val Glu Lys Gly Arg Glu Val Ile Lys Leu
Glu Gly Asp225 230 235
240 Val Val Pro Lys Thr Lys Ile Gln Glu Lys Val Lys Lys Ala Glu Ser
245 250 255 Asp Leu Asn Gln
Lys Phe Lys Glu Ile Thr Gly Leu Lys Gln Asn Leu 260
265 270 Leu Ser Leu Lys Ile Asp Asn Leu Tyr
Thr Ser Ala Ser Ile Asp Arg 275 280
285 Thr Glu Thr Val Ile Lys Gln Leu Val Ser Asn Val Pro Asn
Asn Leu 290 295 300
Leu Leu Asn Ile Met Lys Asn Met Asn Asn Lys Thr Leu Phe Thr Ile305
310 315 320 Thr Asp Lys Ile Leu
Ile Pro Gly Lys Glu Gly Val Leu Gly Tyr Tyr 325
330 335 Asp Thr Ile Ser Lys Thr Leu Phe Ile Gln
Ile Asp His Leu Gly His 340 345
350 Lys Asn Asn Glu Gly Asn Asp Thr Asn Thr Leu Leu His Glu Phe
Gly 355 360 365 His
Ala Val Asp His Leu Ala Lys Gly Glu Ile Gln Ser Lys Ser Ser 370
375 380 Lys Phe Ile Glu Ile Phe
Asn Arg Glu Arg Gly Asn Ile Thr Ile Glu385 390
395 400 Pro Tyr Ile Lys Gln Asp Ala Ala Glu Phe Phe
Ala Gly Val Phe Asn 405 410
415 Tyr Leu Tyr Ser Pro Lys Ile Ser Asp Arg Glu Gln Ile Gln Lys Glu
420 425 430 Ala Pro Asp
Ala Cys Lys Phe Ile Arg Asn Leu Ile His Gly Leu His 435
440 445 321113DNAPaenibacillus
popilliaeCDS(1)...(1113) 32atg aat aaa tta att aaa gta gaa gaa aat aaa
aca cct caa acc caa 48Met Asn Lys Leu Ile Lys Val Glu Glu Asn Lys
Thr Pro Gln Thr Gln1 5 10
15act ata tac act agt ttt aac gca acc gat att ggt ttt gca tcg aat
96Thr Ile Tyr Thr Ser Phe Asn Ala Thr Asp Ile Gly Phe Ala Ser Asn
20 25 30tca gat ata aaa gat gga ttt
cta aat ttt gat gag caa aaa ata aat 144Ser Asp Ile Lys Asp Gly Phe
Leu Asn Phe Asp Glu Gln Lys Ile Asn 35 40
45aca att ata aaa tat tta aaa atg gga aat ttc cca gat ttt cga
gtg 192Thr Ile Ile Lys Tyr Leu Lys Met Gly Asn Phe Pro Asp Phe Arg
Val 50 55 60ggg aat ttg tta cca tcc
gaa cca cat tcc aca gta aat gct ttc ttt 240Gly Asn Leu Leu Pro Ser
Glu Pro His Ser Thr Val Asn Ala Phe Phe65 70
75 80aca caa agg cga att tta ata gag tta gag gtt
ccg gct ggt act tat 288Thr Gln Arg Arg Ile Leu Ile Glu Leu Glu Val
Pro Ala Gly Thr Tyr 85 90
95ctt gcg cat tta gga aac ggt caa acc att ttc cct tta gat tat gga
336Leu Ala His Leu Gly Asn Gly Gln Thr Ile Phe Pro Leu Asp Tyr Gly
100 105 110atg aag tta act gat cag
gcg gga acg att att gga aag caa gta cta 384Met Lys Leu Thr Asp Gln
Ala Gly Thr Ile Ile Gly Lys Gln Val Leu 115 120
125aaa ttg aaa gca ctt gtt gtc ccg aag gat gat att ctg aaa
gaa act 432Lys Leu Lys Ala Leu Val Val Pro Lys Asp Asp Ile Leu Lys
Glu Thr 130 135 140aat gtg cag atg ttc
att tta tac aaa tca ata tcc aat ata ttg cgt 480Asn Val Gln Met Phe
Ile Leu Tyr Lys Ser Ile Ser Asn Ile Leu Arg145 150
155 160tct aag gga ttt gat gaa aag gat ata gaa
agc ctg aaa gct cag tgc 528Ser Lys Gly Phe Asp Glu Lys Asp Ile Glu
Ser Leu Lys Ala Gln Cys 165 170
175atg ttt ata ttt tca ggc cct aat gta ttg ttg gca ata gaa aat tct
576Met Phe Ile Phe Ser Gly Pro Asn Val Leu Leu Ala Ile Glu Asn Ser
180 185 190caa agt gca atg ctt gat
tta cta act aat gag tat ata cca aat aat 624Gln Ser Ala Met Leu Asp
Leu Leu Thr Asn Glu Tyr Ile Pro Asn Asn 195 200
205tta tta aga gat aca ttg tta aaa tta aaa caa cac gcg ggg
att gct 672Leu Leu Arg Asp Thr Leu Leu Lys Leu Lys Gln His Ala Gly
Ile Ala 210 215 220ttt cta agt gtg cct
att tgt atg gat aaa gcg att gcc ggg agt aca 720Phe Leu Ser Val Pro
Ile Cys Met Asp Lys Ala Ile Ala Gly Ser Thr225 230
235 240tca ttt cct aaa aat ggc gat aaa cct aat
atg agt ata atc cca acc 768Ser Phe Pro Lys Asn Gly Asp Lys Pro Asn
Met Ser Ile Ile Pro Thr 245 250
255cat caa tct tta ctg agc cag ttg gat gaa cat ata agt aca tca cgt
816His Gln Ser Leu Leu Ser Gln Leu Asp Glu His Ile Ser Thr Ser Arg
260 265 270aca tta cat cat gaa ttt
ggt cat gta ata gat cga gaa att cta aat 864Thr Leu His His Glu Phe
Gly His Val Ile Asp Arg Glu Ile Leu Asn 275 280
285ggg att tct tcc act cca gag ttt aaa gcg ctg ttt gaa aaa
gaa aaa 912Gly Ile Ser Ser Thr Pro Glu Phe Lys Ala Leu Phe Glu Lys
Glu Lys 290 295 300aat aat att aca gaa
ata aat acg tat gcc aac tat gca aaa acg aat 960Asn Asn Ile Thr Glu
Ile Asn Thr Tyr Ala Asn Tyr Ala Lys Thr Asn305 310
315 320tca caa gaa ttt ttt gca gag gtt ttt aaa
tct atg gtt tcc atg ggg 1008Ser Gln Glu Phe Phe Ala Glu Val Phe Lys
Ser Met Val Ser Met Gly 325 330
335aat gag aaa tat cca tca agt tat tat cgt gat tct att gag aaa gaa
1056Asn Glu Lys Tyr Pro Ser Ser Tyr Tyr Arg Asp Ser Ile Glu Lys Glu
340 345 350gct cct gag act gtg aga
ttt ata aaa gat aaa ttg aaa gag aaa gga 1104Ala Pro Glu Thr Val Arg
Phe Ile Lys Asp Lys Leu Lys Glu Lys Gly 355 360
365tat gta ctt
1113Tyr Val Leu 37033371PRTPaenibacillus popilliae 33Met Asn
Lys Leu Ile Lys Val Glu Glu Asn Lys Thr Pro Gln Thr Gln1 5
10 15 Thr Ile Tyr Thr Ser Phe Asn
Ala Thr Asp Ile Gly Phe Ala Ser Asn 20 25
30 Ser Asp Ile Lys Asp Gly Phe Leu Asn Phe Asp Glu
Gln Lys Ile Asn 35 40 45
Thr Ile Ile Lys Tyr Leu Lys Met Gly Asn Phe Pro Asp Phe Arg Val
50 55 60 Gly Asn Leu
Leu Pro Ser Glu Pro His Ser Thr Val Asn Ala Phe Phe65 70
75 80 Thr Gln Arg Arg Ile Leu Ile Glu
Leu Glu Val Pro Ala Gly Thr Tyr 85 90
95 Leu Ala His Leu Gly Asn Gly Gln Thr Ile Phe Pro Leu
Asp Tyr Gly 100 105 110
Met Lys Leu Thr Asp Gln Ala Gly Thr Ile Ile Gly Lys Gln Val Leu
115 120 125 Lys Leu Lys Ala
Leu Val Val Pro Lys Asp Asp Ile Leu Lys Glu Thr 130
135 140 Asn Val Gln Met Phe Ile Leu Tyr
Lys Ser Ile Ser Asn Ile Leu Arg145 150
155 160 Ser Lys Gly Phe Asp Glu Lys Asp Ile Glu Ser Leu
Lys Ala Gln Cys 165 170
175 Met Phe Ile Phe Ser Gly Pro Asn Val Leu Leu Ala Ile Glu Asn Ser
180 185 190 Gln Ser Ala
Met Leu Asp Leu Leu Thr Asn Glu Tyr Ile Pro Asn Asn 195
200 205 Leu Leu Arg Asp Thr Leu Leu Lys
Leu Lys Gln His Ala Gly Ile Ala 210 215
220 Phe Leu Ser Val Pro Ile Cys Met Asp Lys Ala Ile Ala
Gly Ser Thr225 230 235
240 Ser Phe Pro Lys Asn Gly Asp Lys Pro Asn Met Ser Ile Ile Pro Thr
245 250 255 His Gln Ser Leu
Leu Ser Gln Leu Asp Glu His Ile Ser Thr Ser Arg 260
265 270 Thr Leu His His Glu Phe Gly His Val
Ile Asp Arg Glu Ile Leu Asn 275 280
285 Gly Ile Ser Ser Thr Pro Glu Phe Lys Ala Leu Phe Glu Lys
Glu Lys 290 295 300
Asn Asn Ile Thr Glu Ile Asn Thr Tyr Ala Asn Tyr Ala Lys Thr Asn305
310 315 320 Ser Gln Glu Phe Phe
Ala Glu Val Phe Lys Ser Met Val Ser Met Gly 325
330 335 Asn Glu Lys Tyr Pro Ser Ser Tyr Tyr Arg
Asp Ser Ile Glu Lys Glu 340 345
350 Ala Pro Glu Thr Val Arg Phe Ile Lys Asp Lys Leu Lys Glu Lys
Gly 355 360 365 Tyr
Val Leu 370 34987DNABacillus thuringiensisCDS(1)...(987) 34atg gtg
aga gac tat cct gat ttt gat ggg atg ata aga aaa gcc gct 48Met Val
Arg Asp Tyr Pro Asp Phe Asp Gly Met Ile Arg Lys Ala Ala1 5
10 15cga aag tgg gca gaa gca aac aga
ttg aca ttc caa gat att tcg tat 96Arg Lys Trp Ala Glu Ala Asn Arg
Leu Thr Phe Gln Asp Ile Ser Tyr 20 25
30gca gat cct tta acc aat tca gat aca atc agt cta aat gcc aaa
ttc 144Ala Asp Pro Leu Thr Asn Ser Asp Thr Ile Ser Leu Asn Ala Lys
Phe 35 40 45aaa gat atc gga tgc
ccc gaa gaa tgt gtc gaa tta gaa aaa ata agc 192Lys Asp Ile Gly Cys
Pro Glu Glu Cys Val Glu Leu Glu Lys Ile Ser 50 55
60gtc gca caa gct ttt acg aat aat acg ggg caa caa caa aaa
gaa aca 240Val Ala Gln Ala Phe Thr Asn Asn Thr Gly Gln Gln Gln Lys
Glu Thr65 70 75 80ttt
aca acc gca aca ttt gta gaa gat gag tat act tgg gag aat gac 288Phe
Thr Thr Ala Thr Phe Val Glu Asp Glu Tyr Thr Trp Glu Asn Asp
85 90 95tat cat ttt gtg ctt cca gga
caa aac ttc ctt ata atg ccc cgt ctt 336Tyr His Phe Val Leu Pro Gly
Gln Asn Phe Leu Ile Met Pro Arg Leu 100 105
110cct cgg tca gct cat agg gat att aat cca ggt ttc ctt gtg
aat ttc 384Pro Arg Ser Ala His Arg Asp Ile Asn Pro Gly Phe Leu Val
Asn Phe 115 120 125ttt ggt caa aat
caa caa ttt cat act aaa atg aga gat cta cgt cca 432Phe Gly Gln Asn
Gln Gln Phe His Thr Lys Met Arg Asp Leu Arg Pro 130
135 140att aac ggg gaa gta ttt tta gaa cca tct agc agt
gca aca att caa 480Ile Asn Gly Glu Val Phe Leu Glu Pro Ser Ser Ser
Ala Thr Ile Gln145 150 155
160ctg caa gtg gaa aaa caa tat att tct caa ccg tat gaa att gaa tta
528Leu Gln Val Glu Lys Gln Tyr Ile Ser Gln Pro Tyr Glu Ile Glu Leu
165 170 175tcg ata cta gga agt
att att gtg ata gca cga gat ggg cag aac cga 576Ser Ile Leu Gly Ser
Ile Ile Val Ile Ala Arg Asp Gly Gln Asn Arg 180
185 190agc agt gag aac tat gtt caa tta aca gat ctc atg
cca ctt ctc tgc 624Ser Ser Glu Asn Tyr Val Gln Leu Thr Asp Leu Met
Pro Leu Leu Cys 195 200 205cct tgt
aaa aac ttt ttt tgt aga ggg cga gcg ttg gta ttc gtt gaa 672Pro Cys
Lys Asn Phe Phe Cys Arg Gly Arg Ala Leu Val Phe Val Glu 210
215 220cag gga acg ttc aag gga gta ttg agc cga gcg
ata cgt gca tac gtc 720Gln Gly Thr Phe Lys Gly Val Leu Ser Arg Ala
Ile Arg Ala Tyr Val225 230 235
240aca caa acg ctt cac aaa gac gga aaa acg cta gaa tat gaa att cct
768Thr Gln Thr Leu His Lys Asp Gly Lys Thr Leu Glu Tyr Glu Ile Pro
245 250 255tta aac gca tca cct
gac agg gaa agc gaa ttc tcc cca caa cct ttg 816Leu Asn Ala Ser Pro
Asp Arg Glu Ser Glu Phe Ser Pro Gln Pro Leu 260
265 270gca gca aga tgt atc tca gac gaa gaa tcg aat ggg
aac cct tcg att 864Ala Ala Arg Cys Ile Ser Asp Glu Glu Ser Asn Gly
Asn Pro Ser Ile 275 280 285ttg tca
tcg aga cca tcg aat cct aca gcc tat tcc cag caa cct atg 912Leu Ser
Ser Arg Pro Ser Asn Pro Thr Ala Tyr Ser Gln Gln Pro Met 290
295 300aca act gac tct aca tcc tgt gga tgt tct tct
tgt atg tct gaa aaa 960Thr Thr Asp Ser Thr Ser Cys Gly Cys Ser Ser
Cys Met Ser Glu Lys305 310 315
320tca aat aac aat cta tat ata aat cag
987Ser Asn Asn Asn Leu Tyr Ile Asn Gln
32535329PRTBacillus thuringiensis 35Met Val Arg Asp Tyr Pro Asp Phe Asp
Gly Met Ile Arg Lys Ala Ala1 5 10
15 Arg Lys Trp Ala Glu Ala Asn Arg Leu Thr Phe Gln Asp Ile
Ser Tyr 20 25 30
Ala Asp Pro Leu Thr Asn Ser Asp Thr Ile Ser Leu Asn Ala Lys Phe 35
40 45 Lys Asp Ile Gly Cys
Pro Glu Glu Cys Val Glu Leu Glu Lys Ile Ser 50 55
60 Val Ala Gln Ala Phe Thr Asn Asn Thr Gly
Gln Gln Gln Lys Glu Thr65 70 75
80 Phe Thr Thr Ala Thr Phe Val Glu Asp Glu Tyr Thr Trp Glu Asn
Asp 85 90 95 Tyr
His Phe Val Leu Pro Gly Gln Asn Phe Leu Ile Met Pro Arg Leu
100 105 110 Pro Arg Ser Ala His
Arg Asp Ile Asn Pro Gly Phe Leu Val Asn Phe 115
120 125 Phe Gly Gln Asn Gln Gln Phe His Thr
Lys Met Arg Asp Leu Arg Pro 130 135
140 Ile Asn Gly Glu Val Phe Leu Glu Pro Ser Ser Ser Ala
Thr Ile Gln145 150 155
160 Leu Gln Val Glu Lys Gln Tyr Ile Ser Gln Pro Tyr Glu Ile Glu Leu
165 170 175 Ser Ile Leu Gly
Ser Ile Ile Val Ile Ala Arg Asp Gly Gln Asn Arg 180
185 190 Ser Ser Glu Asn Tyr Val Gln Leu Thr
Asp Leu Met Pro Leu Leu Cys 195 200
205 Pro Cys Lys Asn Phe Phe Cys Arg Gly Arg Ala Leu Val Phe
Val Glu 210 215 220
Gln Gly Thr Phe Lys Gly Val Leu Ser Arg Ala Ile Arg Ala Tyr Val225
230 235 240 Thr Gln Thr Leu His
Lys Asp Gly Lys Thr Leu Glu Tyr Glu Ile Pro 245
250 255 Leu Asn Ala Ser Pro Asp Arg Glu Ser Glu
Phe Ser Pro Gln Pro Leu 260 265
270 Ala Ala Arg Cys Ile Ser Asp Glu Glu Ser Asn Gly Asn Pro Ser
Ile 275 280 285 Leu
Ser Ser Arg Pro Ser Asn Pro Thr Ala Tyr Ser Gln Gln Pro Met 290
295 300 Thr Thr Asp Ser Thr Ser
Cys Gly Cys Ser Ser Cys Met Ser Glu Lys305 310
315 320 Ser Asn Asn Asn Leu Tyr Ile Asn Gln
325 361071DNABacillus thuringiensisCDS(1)...(1071)
36atg aaa aag tct tgt aat ccc aag aaa gta aat cct aac gaa gtg aat
48Met Lys Lys Ser Cys Asn Pro Lys Lys Val Asn Pro Asn Glu Val Asn1
5 10 15cca aca gaa ttt att tta
tac gac ttg gat aac tat tta aat cat att 96Pro Thr Glu Phe Ile Leu
Tyr Asp Leu Asp Asn Tyr Leu Asn His Ile 20 25
30ttg cat aat gta cag tac gac cct gtg aat ctg tcc agt
atg tac gac 144Leu His Asn Val Gln Tyr Asp Pro Val Asn Leu Ser Ser
Met Tyr Asp 35 40 45ggt tat aag
cca tca ggt ggt tgg aat ttt gat gga aaa ttc tat cct 192Gly Tyr Lys
Pro Ser Gly Gly Trp Asn Phe Asp Gly Lys Phe Tyr Pro 50
55 60acc cca act tct agc caa ata cct aac gct tgc gta
cca tac ggt aca 240Thr Pro Thr Ser Ser Gln Ile Pro Asn Ala Cys Val
Pro Tyr Gly Thr65 70 75
80tac aaa cct tat tgt aaa tat gaa cca tat caa agt ggt act tgg gtt
288Tyr Lys Pro Tyr Cys Lys Tyr Glu Pro Tyr Gln Ser Gly Thr Trp Val
85 90 95gta gat aca tcc gct tta
atc gca gaa ggg aca gat ata tat gaa gaa 336Val Asp Thr Ser Ala Leu
Ile Ala Glu Gly Thr Asp Ile Tyr Glu Glu 100
105 110atc gaa tcc atg gct cta gaa acc cct att att gcg
gat aga cat gaa 384Ile Glu Ser Met Ala Leu Glu Thr Pro Ile Ile Ala
Asp Arg His Glu 115 120 125tat aat
aat aac tca tct tta gaa caa tcc tat gtg acc cct gcc tat 432Tyr Asn
Asn Asn Ser Ser Leu Glu Gln Ser Tyr Val Thr Pro Ala Tyr 130
135 140tca gaa aca acg act aca act acg aca aat aca
aca aca cac gga tgt 480Ser Glu Thr Thr Thr Thr Thr Thr Thr Asn Thr
Thr Thr His Gly Cys145 150 155
160aaa gta aat cca aaa atc agt tat tcg cgt aaa tcg aaa tat aaa gtt
528Lys Val Asn Pro Lys Ile Ser Tyr Ser Arg Lys Ser Lys Tyr Lys Val
165 170 175ggt att aag gat aca
gaa aat gga ttt aac ctg gaa tta ggg gca gaa 576Gly Ile Lys Asp Thr
Glu Asn Gly Phe Asn Leu Glu Leu Gly Ala Glu 180
185 190tat aac ttc agc aat aca aac tca aac acg gct acg
aca act cgg act 624Tyr Asn Phe Ser Asn Thr Asn Ser Asn Thr Ala Thr
Thr Thr Arg Thr 195 200 205gtg aca
ttt cca tca ttt acg aca aaa gtg cca ccc tat acc act acg 672Val Thr
Phe Pro Ser Phe Thr Thr Lys Val Pro Pro Tyr Thr Thr Thr 210
215 220att gtg acg gtc ata tta aac aaa gga aca tat
gcc aat tat aat gtt 720Ile Val Thr Val Ile Leu Asn Lys Gly Thr Tyr
Ala Asn Tyr Asn Val225 230 235
240cca gtc caa acg aat tta ttt ggg aga ttt ctc act cat cac ggc caa
768Pro Val Gln Thr Asn Leu Phe Gly Arg Phe Leu Thr His His Gly Gln
245 250 255ggc atg gat cca aac
gac tcg cgt acg tac aat tat ttt gac cta tat 816Gly Met Asp Pro Asn
Asp Ser Arg Thr Tyr Asn Tyr Phe Asp Leu Tyr 260
265 270cct tgc gta gaa tta aat caa aca tgc tgt gca gca
tca tct tgt agc 864Pro Cys Val Glu Leu Asn Gln Thr Cys Cys Ala Ala
Ser Ser Cys Ser 275 280 285gaa tgt
gta aca gac atg gtg caa gct ctt cct gac aat ctt acg gtt 912Glu Cys
Val Thr Asp Met Val Gln Ala Leu Pro Asp Asn Leu Thr Val 290
295 300cgt ttt aat gga aca ggg tca ttt ata gcg gat
gtt gca tcg aat aac 960Arg Phe Asn Gly Thr Gly Ser Phe Ile Ala Asp
Val Ala Ser Asn Asn305 310 315
320ttt gtc att act act gaa gag gtt gac aat gca aca gga gta acc att
1008Phe Val Ile Thr Thr Glu Glu Val Asp Asn Ala Thr Gly Val Thr Ile
325 330 335tcg aaa aaa aca gaa
tac gta ccc gct att tac gga ccc ccg acc aca 1056Ser Lys Lys Thr Glu
Tyr Val Pro Ala Ile Tyr Gly Pro Pro Thr Thr 340
345 350aca gta acc aca tct
1071Thr Val Thr Thr Ser 35537357PRTBacillus
thuringiensis 37Met Lys Lys Ser Cys Asn Pro Lys Lys Val Asn Pro Asn Glu
Val Asn1 5 10 15
Pro Thr Glu Phe Ile Leu Tyr Asp Leu Asp Asn Tyr Leu Asn His Ile
20 25 30 Leu His Asn Val Gln
Tyr Asp Pro Val Asn Leu Ser Ser Met Tyr Asp 35 40
45 Gly Tyr Lys Pro Ser Gly Gly Trp Asn Phe
Asp Gly Lys Phe Tyr Pro 50 55 60
Thr Pro Thr Ser Ser Gln Ile Pro Asn Ala Cys Val Pro Tyr Gly
Thr65 70 75 80 Tyr
Lys Pro Tyr Cys Lys Tyr Glu Pro Tyr Gln Ser Gly Thr Trp Val
85 90 95 Val Asp Thr Ser Ala Leu
Ile Ala Glu Gly Thr Asp Ile Tyr Glu Glu 100
105 110 Ile Glu Ser Met Ala Leu Glu Thr Pro Ile
Ile Ala Asp Arg His Glu 115 120
125 Tyr Asn Asn Asn Ser Ser Leu Glu Gln Ser Tyr Val Thr Pro
Ala Tyr 130 135 140
Ser Glu Thr Thr Thr Thr Thr Thr Thr Asn Thr Thr Thr His Gly Cys145
150 155 160 Lys Val Asn Pro Lys
Ile Ser Tyr Ser Arg Lys Ser Lys Tyr Lys Val 165
170 175 Gly Ile Lys Asp Thr Glu Asn Gly Phe Asn
Leu Glu Leu Gly Ala Glu 180 185
190 Tyr Asn Phe Ser Asn Thr Asn Ser Asn Thr Ala Thr Thr Thr Arg
Thr 195 200 205 Val
Thr Phe Pro Ser Phe Thr Thr Lys Val Pro Pro Tyr Thr Thr Thr 210
215 220 Ile Val Thr Val Ile Leu
Asn Lys Gly Thr Tyr Ala Asn Tyr Asn Val225 230
235 240 Pro Val Gln Thr Asn Leu Phe Gly Arg Phe Leu
Thr His His Gly Gln 245 250
255 Gly Met Asp Pro Asn Asp Ser Arg Thr Tyr Asn Tyr Phe Asp Leu Tyr
260 265 270 Pro Cys Val
Glu Leu Asn Gln Thr Cys Cys Ala Ala Ser Ser Cys Ser 275
280 285 Glu Cys Val Thr Asp Met Val Gln
Ala Leu Pro Asp Asn Leu Thr Val 290 295
300 Arg Phe Asn Gly Thr Gly Ser Phe Ile Ala Asp Val Ala
Ser Asn Asn305 310 315
320 Phe Val Ile Thr Thr Glu Glu Val Asp Asn Ala Thr Gly Val Thr Ile
325 330 335 Ser Lys Lys Thr
Glu Tyr Val Pro Ala Ile Tyr Gly Pro Pro Thr Thr 340
345 350 Thr Val Thr Thr Ser 355
38292PRTBacillus sphaericus 38Met Lys Arg Thr Lys Leu Leu Phe Tyr Ile
Met Ile Ala Ser Phe Leu1 5 10
15 Phe Val Asn Gly Ser Ile Tyr Thr Ala Lys Ala Thr Thr Ile Asp
Glu 20 25 30 Asn
Asn Leu Asp Ile Ile Lys Gln Gln Gly Val Ser Ile Glu Asp Ile 35
40 45 Asp Arg Lys Ile Asp Asn
Met Ile Ala Ser Ile Pro Pro Leu Phe Gly 50 55
60 Phe Leu Pro Tyr Ser Arg Phe Pro Tyr Ile Phe
Gly Glu Ser Val Asp65 70 75
80 Val Ser Gly Ile Asn Ile Glu Asn Thr Asn Val Thr Ser Val Val Pro
85 90 95 Leu Phe Ile
Gly Ser Asn Thr Phe Glu Asn Thr Thr Asp Arg Thr Met 100
105 110 Thr Phe Asn Thr Val Ser Phe Ser
Lys Ser Ile Thr Asp Ser Thr Thr 115 120
125 Thr Gln Thr Leu Asn Gly Phe Lys Thr Ala Phe Glu Ala
Ser Gly Lys 130 135 140
Val Gly Ile Pro Leu Val Ala Glu Gly Gln Ile Lys Thr Thr Leu Glu145
150 155 160 Tyr Asn Phe Ser His
Thr Asn Ser Asn Thr Thr Ser Val Thr Thr Thr 165
170 175 Tyr Thr Val Pro Pro Gln Pro Ile Pro Val
Pro Pro His Thr Lys Thr 180 185
190 Arg Thr Asp Val Tyr Leu Asn Gln Val Ser Ile Ser Gly Asn Val
Glu 195 200 205 Ile
Tyr Ala Asp Ala Ile Thr Gly Ile Lys Ala Glu Ser Ser Gly Thr 210
215 220 Val Ile Ser Ile Gly Asp
Gly Leu Asn Leu Ala Ser Asn Thr Tyr Gly225 230
235 240 Leu Ile Arg Ser Pro Gln Asp Pro Asp Arg Val
Arg Ala Ile Gly Ser 245 250
255 Gly Lys Phe Asn Leu Ile His Gly Ala Asp Phe Thr Ala Ile Thr Tyr
260 265 270 Asp Ile Thr
Ser Gly Glu Ala Ser Ala Arg Ile Ile Asp Val Lys Glu 275
280 285 Ile Ser Phe Lys 290
39362PRTBacillus sphaericus 39Ser Phe Ile Pro Thr Glu Gly Lys Tyr Ile Arg
Val Met Asp Phe Tyr1 5 10
15 Asn Ser Glu Tyr Pro Phe Cys Ile His Ala Pro Ser Ala Pro Asn Gly
20 25 30 Asp Ile Met
Thr Glu Ile Cys Ser Arg Glu Asn Asn Gln Tyr Phe Ile 35
40 45 Phe Phe Pro Thr Asp Asp Gly Arg
Val Ile Ile Ala Asn Arg His Asn 50 55
60 Gly Ser Val Phe Thr Gly Glu Ala Thr Ser Val Val Ser
Asp Ile Tyr65 70 75 80
Thr Gly Ser Pro Ser Gln Phe Phe Arg Glu Val Lys Arg Thr Met Ser
85 90 95 Thr Tyr Tyr Leu Ala
Ile Gln Asn Pro Glu Ser Ala Thr Asp Val Arg 100
105 110 Ala Leu Glu Pro Asn Ser His Glu Leu Pro
Ser Arg Leu Tyr Phe Thr 115 120
125 Asn Asn Ile Glu Asn Asn Ser Asn Ile Leu Ile Ser Asn Lys
Glu Gln 130 135 140
Ile Tyr Leu Thr Leu Pro Ser Leu Pro Glu Asn Glu Gln Tyr Pro Lys145
150 155 160 Thr Pro Val Leu Ser
Gly Ile Asp Asp Ile Gly Pro Asn Gln Ser Glu 165
170 175 Lys Ser Ile Ile Gly Ser Thr Leu Ile Pro
Cys Ile Met Val Ser Asp 180 185
190 Phe Ile Ser Leu Gly Glu Arg Met Lys Thr Thr Pro Tyr Tyr Tyr
Val 195 200 205 Lys
His Thr Gln Tyr Trp Gln Ser Met Trp Ser Ala Leu Phe Pro Pro 210
215 220 Gly Ser Lys Glu Thr Lys
Thr Glu Lys Ser Gly Ile Thr Asp Thr Ser225 230
235 240 Gln Ile Ser Met Thr Asp Gly Ile Asn Val Ser
Ile Gly Ala Asp Phe 245 250
255 Gly Leu Lys Phe Gly Asn Lys Thr Phe Gly Ile Lys Gly Gly Phe Thr
260 265 270 Tyr Asp Thr
Lys Thr Gln Ile Thr Asn Thr Ser Gln Leu Leu Ile Glu 275
280 285 Thr Thr Tyr Thr Arg Glu Tyr Thr
Asn Thr Glu Asn Phe Pro Val Arg 290 295
300 Tyr Thr Gly Tyr Val Leu Ala Ser Glu Phe Thr Leu His
Arg Ser Asp305 310 315
320 Gly Thr Gln Val Asn Thr Ile Pro Trp Val Ala Leu Asn Asp Asn Tyr
325 330 335 Thr Thr Ile Ala
Arg Tyr Pro His Phe Ala Ser Glu Pro Leu Leu Gly 340
345 350 Asn Thr Lys Ile Ile Thr Asp Asp Gln
Asn 355 360 40327PRTBacillus sphaericus
40Met Met Lys Asn Lys Ala Lys Val Ile Leu Met Gly Ala Thr Ile Gly1
5 10 15 Leu Ser Leu Leu
Ser Ser Pro Ile Ala Met Ala Ala Asn Gly Asp Ser 20
25 30 Asn Val Lys Glu Asn Gln Ser Ile Ala
Asn Phe Ser Pro Val Lys Asn 35 40
45 Ser Phe Pro Asp Ala Ala Asn Gly Ser Arg Phe Leu Val Asn
Tyr Tyr 50 55 60
Gly Arg Tyr Leu Thr Ser Asn Gly Leu Gly Ser Ile Gly Lys His Pro65
70 75 80 Glu Asn Ile Asp Phe
Glu Val Lys Asn Thr Tyr Gly Lys Leu Ser Met 85
90 95 Glu Pro Gln Val Ile Ser Gln Asn Pro Leu
Trp Ala Gly Gln Ser Asp 100 105
110 Leu Arg Asn Asp Thr Asp Arg Asp Gln Thr Leu Ser Ser Gln Glu
Phe 115 120 125 Arg
Lys Ser Phe Ser Asn Thr Thr Thr Ala Thr Thr Glu His Gly Phe 130
135 140 Met Phe Gly Thr Glu Thr
Ser Leu Ala Thr Gly Ile Pro Phe Leu Ala145 150
155 160 Glu Gly Lys Ile Thr Leu Lys Ala Glu Tyr Asn
Phe Ser Ser Ser Gln 165 170
175 Ala Asn Glu Thr Ser Glu Thr Val Glu Tyr Val Ala Pro Ser Gln Ser
180 185 190 Ile Val Val
Pro Pro His Thr Ile Ala Arg Val Val Ala Val Leu Glu 195
200 205 Ile Lys Lys Ile Lys Gly Glu Met
Asp Ile Tyr Ala Glu Val Gly Leu 210 215
220 Asn Lys Glu Lys Phe Gly Tyr Glu Glu Leu Pro Ile Ser
Ser Met Gly225 230 235
240 Gly Leu Lys Trp Val Ser Leu Gly Ser Ile Tyr Glu Glu Ala Tyr Asn
245 250 255 Gln Ala Lys Leu
Ser Gly Thr His Glu Phe Pro Asp Ile Lys Ile Ile 260
265 270 Ser Arg Ser Val Asn Asn Pro Asp Tyr
Phe Leu Ala Ser Gly Lys Gly 275 280
285 Arg Phe Glu Ser Glu Tyr Gly Ser Leu Phe Asn Val Gln Val
Glu Tyr 290 295 300
Ile Ser Thr Lys Ser Asn Glu Val Ile Lys Thr Glu Asn Leu Met Val305
310 315 320 Ser Pro Thr Ile Ile
Ser Glu 325 4179PRTBacillus thuringiensis 41Met
Ser Ala Asp Gly Thr Thr Ser Phe Ser Gln Asn Phe Glu Leu Pro1
5 10 15 Lys Asp Thr Val Pro Ala
Thr Pro Lys Gly Asp Ala Lys Thr Val Glu 20 25
30 Arg Thr Glu Gly Thr Gly Thr Asp Ser Gly Glu
Gly Lys Met Val Glu 35 40 45
Thr Lys Val Gly Ser Pro Leu Asn Asn Glu Lys Val Gln Lys Leu Leu
50 55 60 Lys Glu Gln
Lys Val Gln Val Gln Ile Leu Gly Lys Glu Lys Trp65 70
75 42316PRTBacillus thuringiensis 42Met Tyr
Met Ala Glu Ile Lys Arg Leu Asp Tyr Tyr Leu Gly Ala Leu1 5
10 15 Pro Phe Gly Asn Phe Tyr Val
Asp Asp Cys Asp Thr Leu Lys Asn Phe 20 25
30 Ile Asp Ser Leu Leu Asp Gly Lys Pro Ser Thr Met
Asn Asn Thr Pro 35 40 45
Leu Thr Gly Asn Val Asn Val Thr Asn Gln Ser Val Thr Ile Leu Asp
50 55 60 Asp Leu Asp
Ser Ile Ala Thr Leu Thr Pro Glu Tyr Val Tyr Asp Asn65 70
75 80 Tyr Phe Ser Asn Asp Thr Ser Thr
Glu Lys Thr Tyr Gln Thr Leu Ser 85 90
95 Phe Glu Lys Asp Val Gln Thr Thr Val Ser Thr Thr Val
Thr His Gly 100 105 110
Phe Gln Ile Gly Gly Lys Leu Gly Ala Glu Val Lys Gly Ser Val Ser
115 120 125 Ile Pro Phe Val
Ala Asp Gly Gly Val Thr Val Ser Ala Glu Ile Ser 130
135 140 Gly Gln Tyr Asn Phe Ser Ser Ala
Asp Thr Glu Thr Thr Thr Thr Ser145 150
155 160 Gln Lys Leu Ile Ile Pro Ser Gln Ser Gly Asn Ile
Arg Pro Gly Tyr 165 170
175 Thr Thr Arg Val Gln Ile Met Leu Ala Lys Ile Asn Ile Pro Gln Thr
180 185 190 Ala Val His
Phe Ser Gly Ser Met Ser Gly Thr Val His Arg Asp Pro 195
200 205 Ile Pro Ser Ser Val Ile Gly Leu
Val Asp Tyr Asp Leu Tyr Asp Glu 210 215
220 Val Arg Ser Leu Glu Asn Asn Cys Ser Asn Ser Thr Val
Gly Arg Asp225 230 235
240 Thr Gly Leu Val Leu Asn Asn Ala Asn Gln Ser Val Asp Phe Ser Gly
245 250 255 Ser Gly Phe Phe
Thr Gly Ser Ile Thr Ala Phe Asn Phe Tyr Val Lys 260
265 270 Ile Thr Glu Tyr Pro Ile Asn Asn Ser
Ser Gln Glu Asn Ile Arg Trp 275 280
285 Tyr Ser Ile Glu Pro Lys Val Leu Asn Gln Ser Ile Ile Arg
His Arg 290 295 300
Phe Pro Ser Asn Ser Ser Val Asn Thr Cys Asn Cys305 310
315 43525PRTBacillus thuringiensis 43Met Ala Gln Ser Glu
Phe Asn Gln Asn Leu Arg Glu Gln Gly Gln Ser1 5
10 15 Arg Ala Arg Val Ile Ile Leu Arg Val Asn
Asn Pro Gly Tyr Asn Thr 20 25
30 Asn Thr Leu Asp Ile Ala Asp Ile Glu Asp Ile Ile His Leu Pro
Gln 35 40 45 Ala
Ile Glu Leu Ala Asn Ala Phe Gln Ser Ala Leu Val Pro Thr Thr 50
55 60 Ser Asn Phe Gly Glu Asp
Thr Leu Arg Phe Asp Val Glu Arg Gly Leu65 70
75 80 Gly Ile Ala Thr His Val Tyr Pro Arg Ala Ile
Asn Val Asn Tyr Val 85 90
95 Thr Arg Thr Leu Ser Gln Thr Asn Asn Gln Val Gln Ser Met Ile Asn
100 105 110 Lys Val Ile
Glu Glu Leu Lys Ser Leu Leu Gly Ile Asn Leu Ala Asn 115
120 125 Ser Val Leu Gln Gln Leu Thr Thr
Val Ile Thr Glu Thr Phe Thr Asn 130 135
140 Leu Tyr Val Gln Gln Gln Ser Ala Trp Leu Phe Trp Gly
Arg Gln Thr145 150 155
160 Ser Ser Gln Thr Asn Tyr Thr Tyr Asn Ile Val Phe Ala Ile Gln Asn
165 170 175 Ala Gln Thr Gly
Ser Phe Met Lys Ala Ile Pro Ile Gly Phe Glu Ile 180
185 190 Ser Ala Tyr Ile Ala Arg Glu Arg Leu
Leu Phe Phe Asn Ile Gln Asp 195 200
205 Tyr Ala Ser Tyr Ser Val Lys Ile His Ala Ile Gln Val Met
Gln Pro 210 215 220
Leu Ile His Glu Ser Phe Gln Pro Leu Arg Gly Ile Phe Asn Ile Ile225
230 235 240 Thr Ser Val Asn Asn
Arg Ser Ala Ile Gln Ile Thr Glu Tyr Tyr Asp 245
250 255 Glu Asn Thr His Asp Tyr Pro Val Lys Leu
Trp Asp Tyr Asn Asn Ile 260 265
270 Ile Asn Gln Lys Trp Ile Leu Val Phe Asn Gln Thr Thr Arg Ala
Tyr 275 280 285 Ser
Ile Gln Asn Leu Ile Ala Arg Tyr Leu Val Leu Thr Trp Asp Ser 290
295 300 Thr Pro Gly Ser Asn Lys
Val Phe Ala Ser Thr Asn Arg Trp Asn Asp305 310
315 320 Ser Gln Phe Trp Ile Leu Glu Ser Thr Ala Asp
Gly Ser Ile Phe Leu 325 330
335 Thr Asn Met Lys Asp Thr Gln Phe Val Leu Glu Ile Glu Asn Ser Ser
340 345 350 Thr Thr Asn
Gly Thr Asn Val Ile Val Asn Arg Lys Asn Asn Asn Ala 355
360 365 Gln Gln Lys Phe Tyr Leu Asn Lys
Val Asn Gln Glu Phe Gln Asp Gly 370 375
380 Val Tyr Lys Ile Lys Thr Ala Leu Asn Asn Ser Ser Val
Leu Gln Met385 390 395
400 Ser Glu Asp Tyr Phe Gly Tyr Thr Ser Asp Tyr Phe Val Lys Leu Trp
405 410 415 Thr Asn Asn Asn
Asn Asp Ile Asn Gln Lys Trp Ile Phe Glu Phe Asp 420
425 430 Ser Thr Lys Ser Ala Tyr Gln Ile Lys
Ser Gln Arg Asp Pro Ser Leu 435 440
445 Val Leu Ala Trp Thr Trp Ser Val Pro Thr Val Lys Leu Pro
Ile Pro 450 455 460
Asn Asn Asp Asp His Leu Trp Phe Leu Gln Asn Ala Gly Ser Gly Thr465
470 475 480 Tyr Tyr Phe Val Asn
Met Thr Asp Thr Arg Tyr Val Leu Glu Val Ala 485
490 495 Ser Ser Ser Thr Thr Asn Gly Thr Ile Leu
Thr Ile Asn Lys Arg Asn 500 505
510 Gly Asn Leu Asn Gln Lys Phe Leu Leu Asp Met Ile Asn
515 520 525 44340PRTBacillus cereus 44Met
Ala Ile Met Asn Asp Ile Ala Gln Asp Ala Ala Arg Ala Trp Asp1
5 10 15 Ile Ile Ala Gly Pro Phe
Ile Arg Pro Gly Thr Thr Pro Thr Asn Arg 20 25
30 Gln Leu Phe Asn Tyr Gln Ile Gly Asn Ile Glu
Val Glu Pro Gly Asn 35 40 45
Leu Asn Phe Ser Val Val Pro Glu Leu Asp Phe Ser Val Ser Gln Asp
50 55 60 Leu Phe Asn
Asn Thr Ser Val Gln Gln Ser Gln Thr Ala Ser Phe Asn65 70
75 80 Glu Ser Arg Thr Glu Thr Thr Ser
Thr Ala Val Thr His Gly Val Lys 85 90
95 Ser Gly Val Thr Val Ser Ala Ser Ala Lys Phe Asn Ala
Lys Ile Leu 100 105 110
Val Lys Ser Ile Glu Gln Thr Ile Thr Thr Thr Val Ser Thr Glu Tyr
115 120 125 Asn Phe Ser Ser
Thr Thr Thr Arg Thr Asn Thr Val Thr Arg Gly Trp 130
135 140 Ser Ile Ala Gln Pro Val Leu Val
Pro Pro His Ser Arg Val Thr Ala145 150
155 160 Thr Leu Gln Ile Tyr Lys Gly Asp Phe Thr Val Pro
Val Leu Leu Ser 165 170
175 Leu Arg Val Tyr Gly Gln Thr Gly Thr Leu Ala Gly Asn Pro Ser Phe
180 185 190 Pro Ser Leu
Tyr Ala Ala Thr Tyr Glu Asn Thr Leu Leu Gly Arg Ile 195
200 205 Arg Glu His Ile Ala Pro Pro Ala
Leu Phe Arg Ala Ser Asn Ala Tyr 210 215
220 Ile Ser Asn Gly Val Gln Ala Ile Trp Arg Gly Thr Ala
Thr Thr Arg225 230 235
240 Val Ser Gln Gly Leu Tyr Ser Val Val Arg Ile Asp Glu Arg Pro Leu
245 250 255 Ala Gly Tyr Ser
Gly Glu Thr Arg Thr Tyr Tyr Leu Pro Val Thr Leu 260
265 270 Ser Asn Ser Ser Gln Ile Leu Thr Pro
Gly Ser Leu Gly Ser Glu Ile 275 280
285 Pro Ile Ile Asn Pro Val Pro Asn Ala Ser Cys Lys Lys Glu
Asn Ser 290 295 300
Pro Ile Ile Ile His His Asp Arg Glu Lys His Arg Glu Arg Asp Tyr305
310 315 320 Asp Lys Glu His Ile
Cys His Asp Gln Ala Glu Lys Tyr Glu Arg Asp 325
330 335 Tyr Asp Lys Glu 340
451131PRTBacillus thuringiensis 45Met Asn Gln Lys Asn Tyr Glu Ile Ile Gly
Ala Ser Thr Asn Gly Thr1 5 10
15 Ile Glu Leu Pro Glu Asp Tyr Asn Thr Ile Val Ser Pro Tyr Asp
Ala 20 25 30 Pro
Ala Ser Val Thr Thr Thr Ile Glu Ile Thr Gly Thr Ile Leu Ser 35
40 45 Asp Leu Gly Val Pro Gly
Ala Ser Ser Val Ser Leu Leu Leu Asn Lys 50 55
60 Leu Ile Asn Leu Leu Trp Pro Asn Asp Thr Asn
Thr Val Trp Gly Thr65 70 75
80 Phe Gly Lys Glu Thr Ala Asp Leu Leu Asn Glu Val Leu Ser Pro Asp
85 90 95 Asp Pro Val
Val Lys Asp Ala Asn Thr Ile Leu Lys Gly Ile Asn Gly 100
105 110 Ser Leu Asn Leu Tyr Leu Asn Ala
Leu Glu Ile Trp Lys Lys Asp Pro 115 120
125 Asn Asn Leu Thr Thr Ile Glu Asn Val Thr Asp Tyr Phe
Arg Ser Leu 130 135 140
Asn Val Val Phe Thr His Asp Met Pro Ser Phe Ala Val Pro Gly Tyr145
150 155 160 Glu Thr Lys Leu Leu
Thr Ile Tyr Ala Gln Ala Ala Asn Leu His Leu 165
170 175 Leu Leu Leu Arg Asp Ala Ser Arg Phe Gly
Glu Gly Trp Gly Leu Thr 180 185
190 Gln Glu Ile Ile Asn Thr Asn Tyr Asn Asp Gln Leu Arg Leu Thr
Ala 195 200 205 Glu
Tyr Thr Asp His Cys Val Lys Trp Tyr Asn Ala Gly Leu Glu Lys 210
215 220 Leu Lys Gly Asn Leu Thr
Gly Glu Asn Trp Tyr Thr Tyr Asn Arg Phe225 230
235 240 Arg Arg Glu Met Thr Leu Met Val Leu Asp Val
Val Ala Leu Phe Pro 245 250
255 Asn Tyr Asp Thr Arg Met Tyr Pro Ile Gly Thr Ser Ser Glu Leu Thr
260 265 270 Arg Met Ile
Tyr Thr Asp Pro Ile Ala Tyr Thr Gln Ser Asp Pro Trp 275
280 285 Tyr Lys Ile Thr Ser Leu Ser Phe
Ser Asn Ile Glu Asn Ser Ala Ile 290 295
300 Pro Ser Pro Ser Phe Phe Arg Trp Leu Lys Ser Val Ser
Ile Asn Ser305 310 315
320 Gln Trp Trp Gly Ser Gly Pro Ser Gln Thr Tyr Tyr Trp Val Gly His
325 330 335 Glu Leu Val Tyr
Ser Asn Ser Asn Ser Asn Gln Ser Leu Lys Val Lys 340
345 350 Tyr Gly Asp Pro Asn Ser Phe Ile Glu
Pro Pro Asp Ser Phe Ser Phe 355 360
365 Ser Ser Thr Asp Val Tyr Arg Thr Ile Ser Val Val Arg Asn
Ser Val 370 375 380
Ser Asn Tyr Ile Val Ser Glu Val Arg Phe Asn Ser Ile Ser Ser Thr385
390 395 400 Asn Gln Ile Ser Glu
Glu Ile Tyr Lys His Gln Ser Asn Trp Ser Arg 405
410 415 Gln Glu Thr Lys Asp Ser Ile Thr Glu Leu
Ser Leu Ala Ala Asn Pro 420 425
430 Pro Thr Thr Phe Gly Asn Val Ala Glu Tyr Ser His Arg Leu Ala
Tyr 435 440 445 Ile
Ser Glu Ala Tyr Gln Ser His Asn Pro Ser Lys Tyr Pro Thr Tyr 450
455 460 Ile Pro Val Phe Gly Trp
Thr His Thr Ser Val Arg Tyr Asp Asn Lys465 470
475 480 Ile Phe Pro Asp Lys Ile Thr Gln Ile Pro Ala
Val Lys Ser Ser Ser 485 490
495 Ala Gln Gly Gly Ser Trp Lys Asn Ile Val Lys Gly Pro Gly Phe Thr
500 505 510 Gly Gly Asp
Val Thr Thr Ala Val Ser Pro Ala Thr Val Thr Asp Ile 515
520 525 Ile Lys Ile Gln Val Thr Leu Asp
Pro Asn Ser Leu Ser Gln Lys Tyr 530 535
540 Arg Ala Arg Leu Arg Tyr Ala Ser Asn Ala Phe Val Pro
Ala Thr Leu545 550 555
560 Tyr Thr Asn Thr Ser Ser Asn Tyr Asn Phe Glu Leu Lys Lys Gly Thr
565 570 575 Thr Glu Gln Phe
Thr Thr Tyr Asn Ser Tyr Gln Tyr Val Asp Ile Pro 580
585 590 Gly Ser Ile Gln Phe Asn Asn Thr Ser
Asp Thr Val Ser Val Tyr Leu 595 600
605 His Met Asp Ser Thr Ser Asn Val Asn Val His Val Asp Arg
Ile Glu 610 615 620
Phe Ile Pro Ile Asp Glu Asn Tyr Asp Glu Arg Phe Gln Leu Glu Lys625
630 635 640 Ala Gln Lys Ala Val
Asn Ala Leu Phe Thr Ala Gly Arg Asn Ala Leu 645
650 655 Gln Thr Asp Val Thr Asp Tyr Lys Val Asp
Gln Val Ser Ile Leu Val 660 665
670 Asp Cys Val Ser Gly Glu Leu Tyr Pro Asn Glu Lys Arg Glu Leu
Leu 675 680 685 Ser
Leu Val Lys Tyr Ala Lys Arg Leu Ser Tyr Ser Arg Asn Leu Leu 690
695 700 Leu Asp Pro Thr Phe Asp
Ser Ile Asn Ser Pro Glu Glu Asn Gly Trp705 710
715 720 Tyr Gly Ser Asn Gly Ile Ala Ile Gly Ser Gly
Asn Ile Val Phe Lys 725 730
735 Gly Asn Tyr Leu Ile Phe Ser Gly Thr Asn Asp Glu Gln Tyr Pro Thr
740 745 750 Tyr Leu Tyr
Gln Lys Ile Asp Glu Thr Lys Leu Lys Glu Tyr Thr Arg 755
760 765 Tyr Lys Leu Arg Gly Phe Ile Glu
Ser Ser Gln Asp Leu Glu Ala Tyr 770 775
780 Val Ile Arg Tyr Asp Ala Lys His Gln Thr Met Asp Val
Ser Asn Asn785 790 795
800 Leu Phe Ser Asp Ile Thr Pro Val Asn Ala Cys Gly Glu Pro Asn Arg
805 810 815 Cys Ala Ala Leu
Pro Tyr Leu Asp Glu Asn Pro Arg Leu Glu Cys Ser 820
825 830 Ser Ile Gln Asp Gly Ile Leu Ser Asp
Ser His Ser Phe Ser Leu His 835 840
845 Ile Asp Thr Gly Ser Ile Asp Phe Asn Glu Asn Val Gly Ile
Trp Val 850 855 860
Leu Phe Lys Ile Ser Thr Pro Glu Gly Tyr Ala Arg Phe Gly Asn Leu865
870 875 880 Glu Val Ile Glu Asp
Gly Pro Val Ile Gly Glu Ala Leu Ala Arg Val 885
890 895 Lys Arg Gln Glu Thr Lys Trp Arg Asn Lys
Leu Thr Gln Leu Arg Thr 900 905
910 Glu Thr Gln Ala Ile Tyr Thr Arg Ala Lys Gln Ala Ile Asp Asn
Leu 915 920 925 Phe
Thr Asn Ala Gln Asp Ser His Leu Lys Ile Gly Ala Thr Phe Ala 930
935 940 Ser Ile Val Ala Ala Arg
Lys Ile Val Gln Ser Ile Arg Glu Ala Tyr945 950
955 960 Met Ser Trp Leu Ser Ile Val Pro Gly Val Asn
Tyr Pro Ile Val Thr 965 970
975 Glu Leu Asn Glu Arg Ile Gln Gln Ala Phe Gln Leu Tyr Asp Val Arg
980 985 990 Asn Val Val
Arg Asn Gly Arg Phe Gln Ser Gly Thr Ser Asp Trp Ile 995
1000 1005 Val Thr Ser Asp Val Arg Val Gln
Glu Glu Asn Gly Asn Asn Val Leu 1010 1015
1020 Val Leu Ser Asn Trp Asp Ala Gln Val Leu Gln Cys Met
Thr Leu Tyr1025 1030 1035
1040 Gln Asp Arg Gly Tyr Ile Leu Arg Val Thr Ala Arg Lys Glu Gly Leu
1045 1050 1055 Gly Glu Gly Tyr
Val Thr Ile Thr Asp Glu Glu Gly Asn Thr Asp Gln 1060
1065 1070 Leu Arg Phe Gly Gly Cys Glu Glu Ile
Asp Ala Ser Asn Ser Phe Val 1075 1080
1085 Ser Thr Gly Tyr Met Thr Lys Glu Leu Glu Phe Phe Pro Asp
Thr Glu 1090 1095 1100
Lys Val Arg Ile Glu Ile Gly Glu Thr Glu Gly Thr Phe Gln Val Glu1105
1110 1115 1120 Ser Val Glu Leu Phe
Leu Met Glu Asp Leu Cys 1125 1130
461138PRTBacillus thuringiensis 46Met Asn Leu Asn Asn Leu Asp Gly Tyr Glu
Asp Ser Asn Arg Thr Leu1 5 10
15 Asn Asn Ser Leu Asn Tyr Pro Thr Gln Lys Ala Leu Ser Pro Ser
Leu 20 25 30 Lys
Asn Met Asn Tyr Gln Asp Phe Leu Ser Ile Thr Glu Arg Glu Gln 35
40 45 Pro Glu Ala Leu Ala Ser
Gly Asn Thr Ala Ile Asn Thr Val Val Ser 50 55
60 Val Thr Gly Ala Thr Leu Ser Ala Leu Gly Val
Pro Gly Ala Ser Phe65 70 75
80 Ile Thr Asn Phe Tyr Leu Lys Ile Ala Gly Leu Leu Trp Pro Glu Asn
85 90 95 Gly Lys Ile
Trp Asp Glu Phe Met Thr Glu Val Glu Ala Leu Ile Asp 100
105 110 Gln Lys Ile Glu Glu Tyr Val Arg
Asn Lys Ala Ile Ala Glu Leu Asp 115 120
125 Gly Leu Gly Ser Ala Leu Asp Lys Tyr Gln Lys Ala Leu
Ala Asp Trp 130 135 140
Leu Gly Lys Gln Asp Asp Pro Glu Ala Ile Leu Ser Val Ala Thr Glu145
150 155 160 Phe Arg Ile Ile Asp
Ser Leu Phe Glu Phe Ser Met Pro Ser Phe Lys 165
170 175 Val Thr Gly Tyr Glu Ile Pro Leu Leu Thr
Val Tyr Ala Gln Ala Ala 180 185
190 Asn Leu His Leu Ala Leu Leu Arg Asp Ser Thr Leu Tyr Gly Asp
Lys 195 200 205 Trp
Gly Phe Thr Gln Asn Asn Ile Glu Glu Asn Tyr Asn Arg Gln Lys 210
215 220 Lys Arg Ile Ser Glu Tyr
Ser Asp His Cys Thr Lys Trp Tyr Asn Ser225 230
235 240 Gly Leu Ser Arg Leu Asn Gly Ser Thr Tyr Glu
Gln Trp Ile Asn Tyr 245 250
255 Asn Arg Phe Arg Arg Glu Met Ile Leu Met Ala Leu Asp Leu Val Ala
260 265 270 Val Phe Pro
Phe His Asp Pro Arg Arg Tyr Ser Met Glu Thr Ser Thr 275
280 285 Gln Leu Thr Arg Glu Val Tyr Thr
Asp Pro Val Ser Leu Ser Ile Ser 290 295
300 Asn Pro Asp Ile Gly Pro Ser Phe Ser Gln Met Glu Asn
Thr Ala Ile305 310 315
320 Arg Thr Pro His Leu Val Asp Tyr Leu Asp Glu Leu Tyr Ile Tyr Thr
325 330 335 Ser Lys Tyr Lys
Ala Phe Ser His Glu Ile Gln Pro Asp Leu Phe Tyr 340
345 350 Trp Ser Ala His Lys Val Ser Phe Lys
Lys Ser Glu Gln Ser Asn Leu 355 360
365 Tyr Thr Thr Gly Ile Tyr Gly Lys Thr Ser Gly Tyr Ile Ser
Ser Gly 370 375 380
Ala Tyr Ser Phe His Gly Asn Asp Ile Tyr Arg Thr Leu Ala Ala Pro385
390 395 400 Ser Val Val Val Tyr
Pro Tyr Thr Gln Asn Tyr Gly Val Glu Gln Val 405
410 415 Glu Phe Tyr Gly Val Lys Gly His Val His
Tyr Arg Gly Asp Asn Lys 420 425
430 Tyr Asp Leu Thr Tyr Asp Ser Ile Asp Gln Leu Pro Pro Asp Gly
Glu 435 440 445 Pro
Ile His Glu Lys Tyr Thr His Arg Leu Cys His Ala Thr Ala Ile 450
455 460 Phe Lys Ser Thr Pro Asp
Tyr Asp Asn Ala Thr Ile Pro Ile Phe Ser465 470
475 480 Trp Thr His Arg Ser Ala Glu Tyr Tyr Asn Arg
Ile Tyr Pro Asn Lys 485 490
495 Ile Thr Lys Ile Pro Ala Val Lys Met Tyr Lys Leu Asp Asp Pro Ser
500 505 510 Thr Val Val
Lys Gly Pro Gly Phe Thr Gly Gly Asp Leu Val Lys Arg 515
520 525 Gly Ser Thr Gly Tyr Ile Gly Asp
Ile Lys Ala Thr Val Asn Ser Pro 530 535
540 Leu Ser Gln Lys Tyr Arg Val Arg Val Arg Tyr Ala Thr
Asn Val Ser545 550 555
560 Gly Gln Phe Asn Val Tyr Ile Asn Asp Lys Ile Thr Leu Gln Thr Lys
565 570 575 Phe Gln Asn Thr
Val Glu Thr Ile Gly Glu Gly Lys Asp Leu Thr Tyr 580
585 590 Gly Ser Phe Gly Tyr Ile Glu Tyr Ser
Thr Thr Ile Gln Phe Pro Asp 595 600
605 Glu His Pro Lys Ile Thr Leu His Leu Ser Asp Leu Ser Asn
Asn Ser 610 615 620
Ser Phe Tyr Val Asp Ser Ile Glu Phe Ile Pro Val Asp Val Asn Tyr625
630 635 640 Ala Glu Lys Glu Lys
Leu Glu Lys Ala Gln Lys Ala Val Asn Thr Leu 645
650 655 Phe Thr Glu Gly Arg Asn Ala Leu Gln Lys
Asp Val Thr Asp Tyr Lys 660 665
670 Val Asp Gln Val Ser Ile Leu Val Asp Cys Ile Ser Gly Asp Leu
Tyr 675 680 685 Pro
Asn Glu Lys Arg Glu Leu Gln Asn Leu Val Lys Tyr Ala Lys Arg 690
695 700 Leu Ser Tyr Ser Arg Asn
Leu Leu Leu Asp Pro Thr Phe Asp Ser Ile705 710
715 720 Asn Ser Ser Glu Glu Asn Gly Trp Tyr Gly Ser
Asn Gly Ile Val Ile 725 730
735 Gly Asn Gly Asp Phe Val Phe Lys Gly Asn Tyr Leu Ile Phe Ser Gly
740 745 750 Thr Asn Asp
Thr Gln Tyr Pro Thr Tyr Leu Tyr Gln Lys Ile Asp Glu 755
760 765 Ser Lys Leu Lys Glu Tyr Thr Arg
Tyr Lys Leu Lys Gly Phe Ile Glu 770 775
780 Ser Ser Gln Asp Leu Glu Ala Tyr Val Ile Arg Tyr Asp
Ala Lys His785 790 795
800 Arg Thr Leu Asp Val Ser Asp Asn Leu Leu Pro Asp Ile Leu Pro Glu
805 810 815 Asn Thr Cys Gly
Glu Pro Asn Arg Cys Ala Ala Gln Gln Tyr Leu Asp 820
825 830 Glu Asn Pro Ser Pro Glu Cys Ser Ser
Met Gln Asp Gly Ile Leu Ser 835 840
845 Asp Ser His Ser Phe Ser Leu Asn Ile Asp Thr Gly Ser Ile
Asn His 850 855 860
Asn Glu Asn Leu Gly Ile Trp Val Leu Phe Lys Ile Ser Thr Leu Glu865
870 875 880 Gly Tyr Ala Lys Phe
Gly Asn Leu Glu Val Ile Glu Asp Gly Pro Val 885
890 895 Ile Gly Glu Ala Leu Ala Arg Val Lys Arg
Gln Glu Thr Lys Trp Arg 900 905
910 Asn Lys Leu Ala Gln Leu Thr Thr Glu Thr Gln Ala Ile Tyr Thr
Arg 915 920 925 Ala
Lys Gln Ala Leu Asp Asn Leu Phe Ala Asn Ala Gln Asp Ser His 930
935 940 Leu Lys Arg Asp Val Thr
Phe Ala Glu Ile Ala Ala Ala Arg Lys Ile945 950
955 960 Val Gln Ser Ile Arg Glu Ala Tyr Met Ser Trp
Leu Ser Val Val Pro 965 970
975 Gly Val Asn His Pro Ile Phe Thr Glu Leu Ser Gly Arg Val Gln Arg
980 985 990 Ala Phe Gln
Leu Tyr Asp Val Arg Asn Val Val Arg Asn Gly Arg Phe 995
1000 1005 Leu Asn Gly Leu Ser Asp Trp Ile
Val Thr Ser Asp Val Lys Val Gln 1010 1015
1020 Glu Glu Asn Gly Asn Asn Val Leu Val Leu Asn Asn Trp
Asp Ala Gln1025 1030 1035
1040 Val Leu Gln Asn Val Lys Leu Tyr Gln Asp Arg Gly Tyr Ile Leu His
1045 1050 1055 Val Thr Ala Arg
Lys Ile Gly Ile Gly Glu Gly Tyr Ile Thr Ile Thr 1060
1065 1070 Asp Glu Glu Gly His Thr Asp Gln Leu
Arg Phe Thr Ala Cys Glu Glu 1075 1080
1085 Ile Asp Ala Ser Asn Ala Phe Ile Ser Gly Tyr Ile Thr Lys
Glu Leu 1090 1095 1100
Glu Phe Phe Pro Asp Thr Glu Lys Val His Ile Glu Ile Gly Glu Thr1105
1110 1115 1120 Glu Gly Ile Phe Leu
Val Glu Ser Ile Glu Leu Phe Leu Met Glu Glu 1125
1130 1135 Leu Cys 47326PRTBacillus thuringiensis
47Met Val Arg Glu Tyr Pro Asp Leu Asp Ser Met Ile Arg Glu Ala Ala1
5 10 15 Gln Lys Trp Ser
Glu Asp Asn Gly Leu Gln Phe Gln Lys Val Ser Phe 20
25 30 Lys Asp Pro Phe Thr Asn Arg Asp Thr
Ile Arg Met Ser Val Lys Phe 35 40
45 Lys Asp Ile Gly Cys Pro Glu Glu Cys Leu Glu Thr Glu Thr
Val Lys 50 55 60
Leu Ser Gln Ala Phe Thr Asn Asn Thr Gly Gln Pro Lys Lys Glu Thr65
70 75 80 Val Asn Thr Val Thr
Tyr Val Glu Asn Gln Leu Thr Trp Glu Asn Asp 85
90 95 Phe His Phe Lys Leu Pro Gly Gln Asn Phe
Leu Ile Leu Pro Arg Ile 100 105
110 Pro Gln Ser Val Arg Met Asp Ile Asn Pro Gly Phe Leu Val Asn
Phe 115 120 125 Phe
Gly Asp Asn Glu Leu Phe Ser Thr Asn Met Arg Asp Arg Arg Pro 130
135 140 Ile Gln Ala Asp Val Phe
Val Glu Pro Gly Ser Ser Ala Ala Ile Gln145 150
155 160 Leu Lys Val Glu Lys Leu His Val Thr Gln Pro
Tyr Glu Ile Glu Leu 165 170
175 Ser Ile Leu Gly Ser Ile Ile Val Thr Ala Gln Gly Ala Glu Arg Tyr
180 185 190 Val Asp Val
Thr Asp Leu Leu Pro Phe Leu Cys Leu Ser Lys Asn Leu 195
200 205 Ser Ser Arg Gly Arg Ala Leu Ile
Phe Leu Glu Gln Gly Thr Phe Lys 210 215
220 Gly Ile Leu Asn Arg Lys Ile Arg Gly Tyr Ala Leu Gln
Thr Arg His225 230 235
240 Cys Asp Gly Lys Thr Ile Glu Tyr Glu Ile Pro Leu Asn Asn Arg Pro
245 250 255 Pro Val Ser Ala
Arg Pro Leu Asn Pro Ala Thr Thr Ala Gln Gln Ser 260
265 270 Arg Lys Thr Asn Asp Ser Ser Cys Gly
Cys Ser Ser Asp Arg Pro Pro 275 280
285 Val Gly Ile Tyr Leu Leu Ser Thr Ile Glu Ser Asn Asn Leu
Phe Ser 290 295 300
Ala Thr Glu Ala Asp Glu Gly Cys Arg Phe Ile Met Trp Met Cys Cys305
310 315 320 Leu Phe Val Cys Ile
Lys 325 48335PRTBacillus thuringiensis 48Met Lys Lys
Ser Cys Asp Pro Asn Pro Val Asn Gln Ser Thr Thr Thr1 5
10 15 Thr Phe Asp Leu Asp Asp Phe Leu
Leu Asn Asn Met Phe Asn Val Thr 20 25
30 Leu Gln Pro Ala Asn Ile Trp Tyr Tyr Gln Asp Glu Trp
Pro Tyr Glu 35 40 45
Ser Pro Tyr Val Pro Thr Pro Thr Ser Asp Asp Leu Ser Lys Gly Cys 50
55 60 Trp Phe Asp Ala Tyr
Val Pro Thr Cys Arg Tyr Asp His Ala Pro Gly65 70
75 80 Tyr Thr Ala Asn Thr Ser Gly Leu Met Ala
Glu Gly Thr Asp Leu Thr 85 90
95 Glu Glu Ile Asp Ser Val Ala Tyr Ala Thr Pro Tyr Ile Ala Asp
Ser 100 105 110 Tyr
Thr Phe Thr Asn Asp Gly Pro Ile Thr Gln Glu Tyr Gln Thr Leu 115
120 125 Ala Tyr Glu Gln Ala Val
Glu Thr Ser Thr Ser Asn Thr Thr Thr His 130 135
140 Gly Cys Arg Val Gly Ser Thr Phe Gly Tyr Ser
Arg Asn Ser Thr Phe145 150 155
160 Thr Ala Lys Ile Arg Asp Thr Glu Lys Gly Phe His Leu Asp Val Gly
165 170 175 Ala Glu Tyr
Asp Phe Thr Asn Thr Asn Thr Phe Thr Thr Ser Thr Thr 180
185 190 Thr Asn Val Leu Val Pro Ser Gln
Val Ile Thr Val Pro Ser Tyr Cys 195 200
205 Thr Ala Tyr Val Thr Met Val Leu Asn Lys Ala Thr Tyr
Ala Lys Ala 210 215 220
Asp Val Pro Leu Ile Thr Thr Leu Ser Gly Arg Phe Phe Ile Asp Glu225
230 235 240 Thr Asp Asn Ser Asp
Glu Tyr Phe Asp Ile Tyr Pro Tyr Val Glu Leu 245
250 255 Val Thr Thr Cys Cys Thr Gly Asn Cys Ser
Gln Cys Val Thr Asp Gln 260 265
270 Leu Gln Leu Asp Ala Val Asn Arg Thr Val Ile Phe Asp Gly Leu
Gly 275 280 285 Ser
Phe Glu Ala Asn Ile Ala Ser Asn Glu Leu Ile Val Arg Thr Lys 290
295 300 Leu Val Asp Asn Val Thr
Gly Ala Thr Ile Ser Glu Gln Ala Gly Arg305 310
315 320 Val Pro Val Val Tyr Gly Pro Ser Thr Thr Lys
Val Thr Thr Ser 325 330
335 49875PRTClostridium perfringens 49Met Asn Ile Gln Ile Lys Asn Val Phe
Ser Phe Leu Thr Leu Thr Ala1 5 10
15 Met Ile Ser Gln Thr Leu Ser Tyr Asn Val Tyr Ala Gln Thr
Thr Thr 20 25 30
Gln Asn Asp Thr Asn Gln Lys Glu Glu Ile Thr Asn Glu Asn Thr Leu 35
40 45 Ser Ser Asn Gly Leu
Met Gly Tyr Tyr Phe Ala Asp Glu His Phe Lys 50 55
60 Asp Leu Glu Leu Met Ala Pro Ile Lys Asn
Gly Asp Leu Lys Phe Glu65 70 75
80 Glu Lys Lys Val Asp Lys Leu Leu Thr Glu Asp Asn Ser Ser Ile
Lys 85 90 95 Ser
Ile Arg Trp Thr Gly Arg Ile Ile Pro Ser Glu Asp Gly Glu Tyr
100 105 110 Ile Leu Ser Thr Asp
Arg Asn Asp Val Leu Met Gln Ile Asn Ala Lys 115
120 125 Gly Asp Ile Ala Lys Thr Leu Lys Val
Asn Met Lys Lys Gly Gln Ala 130 135
140 Tyr Asn Ile Arg Ile Glu Ile Gln Asp Lys Asn Leu Gly
Ser Ile Asp145 150 155
160 Asn Leu Ser Val Pro Lys Leu Tyr Trp Glu Leu Asn Gly Asn Lys Thr
165 170 175 Val Ile Pro Glu
Glu Asn Leu Phe Phe Arg Asp Tyr Ser Lys Ile Asp 180
185 190 Glu Asn Asp Pro Phe Ile Pro Asn Asn
Asn Phe Phe Asp Val Arg Phe 195 200
205 Phe Ser Ala Ala Trp Glu Asp Glu Asp Leu Asp Thr Asp Asn
Asp Asn 210 215 220
Ile Pro Asp Ala Tyr Glu Lys Asn Gly Tyr Thr Ile Lys Asp Ser Ile225
230 235 240 Ala Val Lys Trp Asn
Asp Ser Phe Ala Glu Gln Gly Tyr Lys Lys Tyr 245
250 255 Val Ser Ser Tyr Leu Glu Ser Asn Thr Ala
Gly Asp Pro Tyr Thr Asp 260 265
270 Tyr Gln Lys Ala Ser Gly Ser Ile Asp Lys Ala Ile Lys Leu Glu
Ala 275 280 285 Arg
Asp Pro Leu Val Ala Ala Tyr Pro Val Val Gly Val Gly Met Glu 290
295 300 Asn Leu Ile Ile Ser Thr
Asn Glu His Ala Ser Ser Asp Gln Gly Lys305 310
315 320 Thr Val Ser Arg Ala Thr Thr Asn Ser Lys Thr
Asp Ala Asn Thr Val 325 330
335 Gly Val Ser Ile Ser Ala Gly Tyr Gln Asn Gly Phe Thr Gly Asn Ile
340 345 350 Thr Thr Ser
Tyr Ser His Thr Thr Asp Asn Ser Thr Ala Val Gln Asp 355
360 365 Ser Asn Gly Glu Ser Trp Asn Thr
Gly Leu Ser Ile Asn Lys Gly Glu 370 375
380 Ser Ala Tyr Ile Asn Ala Asn Val Arg Tyr Tyr Asn Thr
Gly Thr Ala385 390 395
400 Pro Met Tyr Lys Val Thr Pro Thr Thr Asn Leu Val Leu Asp Gly Glu
405 410 415 Thr Leu Ala Thr
Ile Lys Ala Gln Asp Asn Gln Ile Gly Asn Asn Leu 420
425 430 Ser Pro Asn Glu Thr Tyr Pro Lys Lys
Gly Leu Ser Pro Leu Ala Leu 435 440
445 Asn Thr Met Asp Gln Phe Asn Ala Arg Leu Ile Pro Ile Asn
Tyr Asp 450 455 460
Gln Leu Lys Lys Leu Asp Ser Gly Lys Gln Ile Lys Leu Glu Thr Thr465
470 475 480 Gln Val Ser Gly Asn
Tyr Gly Thr Lys Asn Ser Gln Gly Gln Ile Ile 485
490 495 Thr Glu Gly Asn Ser Trp Ser Asn Tyr Ile
Ser Gln Ile Asp Ser Val 500 505
510 Ser Ala Ser Ile Ile Leu Asp Thr Gly Ser Gln Thr Phe Glu Arg
Arg 515 520 525 Val
Ala Ala Lys Glu Gln Gly Asn Pro Glu Asp Lys Thr Pro Glu Ile 530
535 540 Thr Ile Gly Glu Ala Ile
Lys Lys Ala Phe Ser Ala Thr Lys Asn Gly545 550
555 560 Glu Leu Leu Tyr Phe Asn Gly Ile Pro Ile Asp
Glu Ser Cys Val Glu 565 570
575 Leu Ile Phe Asp Asp Asn Thr Ser Glu Ile Ile Lys Glu Gln Leu Lys
580 585 590 Tyr Leu Asp
Asp Lys Lys Ile Tyr Asn Val Lys Leu Glu Arg Gly Met 595
600 605 Asn Ile Leu Ile Lys Val Pro Ser
Tyr Phe Thr Asn Phe Asp Glu Tyr 610 615
620 Asn Asn Phe Pro Ala Ser Trp Ser Asn Ile Asp Thr Lys
Asn Gln Asp625 630 635
640 Gly Leu Gln Ser Val Ala Asn Lys Leu Ser Gly Glu Thr Lys Ile Ile
645 650 655 Ile Pro Met Ser
Lys Leu Lys Pro Tyr Lys Arg Tyr Val Phe Ser Gly 660
665 670 Tyr Ser Lys Asp Pro Ser Thr Ser Asn
Ser Ile Thr Val Asn Ile Lys 675 680
685 Ser Lys Glu Gln Lys Thr Asp Tyr Leu Val Pro Glu Lys Asp
Tyr Thr 690 695 700
Lys Phe Ser Tyr Glu Phe Glu Thr Thr Gly Lys Asp Ser Ser Asp Ile705
710 715 720 Glu Ile Thr Leu Thr
Ser Ser Gly Val Ile Phe Leu Asp Asn Leu Ser 725
730 735 Ile Thr Glu Leu Asn Ser Thr Pro Glu Ile
Leu Lys Glu Pro Glu Ile 740 745
750 Lys Val Pro Ser Asp Gln Glu Ile Leu Asp Ala His Asn Lys Tyr
Tyr 755 760 765 Ala
Asp Ile Lys Leu Asp Thr Asn Thr Gly Asn Thr Tyr Ile Asp Gly 770
775 780 Ile Tyr Phe Glu Pro Thr
Gln Thr Asn Lys Glu Ala Leu Asp Tyr Ile785 790
795 800 Gln Lys Tyr Arg Val Glu Ala Thr Leu Gln Tyr
Ser Gly Phe Lys Asp 805 810
815 Ile Gly Thr Lys Asp Lys Glu Ile Arg Asn Tyr Leu Gly Asp Gln Asn
820 825 830 Gln Pro Lys
Thr Asn Tyr Ile Asn Phe Arg Ser Tyr Phe Thr Ser Gly 835
840 845 Glu Asn Val Met Thr Tyr Lys Lys
Leu Arg Ile Tyr Ala Val Thr Pro 850 855
860 Asp Asn Arg Glu Leu Leu Val Leu Ser Val Asn865
870 875 50871PRTBrevibacillus laterosporus
50Met Lys Tyr Met Lys Lys Gly Leu Ser Ser Val Val Ile Gly Thr Leu1
5 10 15 Phe Ala Ser Met
Phe Leu Asn Gly Asn Val Asn Ala Val Tyr Ala Asn 20
25 30 Ser Lys Thr Asn Gln Ile Ala Thr Thr
Thr Gln Ala Ser Lys Asp Asn 35 40
45 Gln Ile Asp Arg Glu Gly Leu Leu Gly Tyr Tyr Phe Lys Gly
Lys Asp 50 55 60
Phe Asn Asp Leu Thr Leu Phe Ala Pro Thr Arg Asp Asn Thr Leu Ile65
70 75 80 Tyr Asp Gln Gln Thr
Ala Asn Thr Leu Val Asp Gln Lys His Gln Glu 85
90 95 Tyr His Ser Ile Arg Trp Ile Gly Leu Ile
Gln Ser Ser Ala Thr Gly 100 105
110 Asp Phe Thr Phe Lys Leu Ser Asp Asp Glu Asn Ala Ile Ile Glu
Leu 115 120 125 Asp
Gly Lys Val Ile Ser Glu Lys Gly Asn Asn Lys Gln Ser Val His 130
135 140 Leu Glu Lys Gly Gln Leu
Val Gln Ile Lys Ile Glu Tyr Gln Ser Asp145 150
155 160 Asp Ala Leu His Ile Asp Asn Lys Ile Phe Lys
Glu Leu Lys Leu Phe 165 170
175 Lys Ile Asp Ser Gln Asn His Ser Gln Gln Val Gln Gln Asp Glu Leu
180 185 190 Arg Asn Pro
Glu Phe Asn Lys Lys Glu Thr Gln Val Phe Leu Lys Lys 195
200 205 Ala Ser Lys Thr Asn Leu Phe Thr
Gln Lys Thr Lys Arg Asp Ile Asp 210 215
220 Glu Asp Thr Asp Thr Asp Gly Asp Ser Ile Pro Asp Val
Trp Glu Glu225 230 235
240 Asn Gly Tyr Thr Ile Gln Asn Lys Val Ala Val Lys Trp Asp Asp Ser
245 250 255 Leu Ala Ser Lys
Gly Tyr Gln Lys Phe Thr Ser Asn Pro Leu Glu Ala 260
265 270 His Thr Val Gly Asp Pro Tyr Ser Asp
Tyr Glu Lys Ala Ala Arg Asp 275 280
285 Met Pro Leu Ser Asn Ala Lys Glu Thr Phe Asn Pro Leu Val
Ala Ala 290 295 300
Phe Pro Ser Val Asn Val Ser Leu Glu Lys Val Ile Leu Ser Lys Asn305
310 315 320 Glu Asp Leu Ser His
Ser Val Glu Ser Ser Gln Ser Thr Asn Trp Ser 325
330 335 Tyr Thr Asn Thr Glu Gly Val Asn Val Asn
Ala Gly Trp Ser Gly Leu 340 345
350 Gly Pro Ser Phe Gly Val Ser Val Asn Tyr Gln His Ser Glu Thr
Val 355 360 365 Ala
Asn Glu Trp Gly Ser Ala Thr Asn Asp Gly Thr His Ile Asn Gly 370
375 380 Ala Glu Ser Ala Tyr Leu
Asn Ala Asn Val Arg Tyr Asn Asn Val Gly385 390
395 400 Thr Gly Ala Ile Tyr Glu Thr Lys Pro Thr Thr
Ser Phe Ile Leu Asp 405 410
415 Gly Thr Thr Ile Gly Thr Ile Lys Ala Lys Glu Asn Thr Thr Ala Leu
420 425 430 Thr Ile Leu
Pro Asp Gln Ser Tyr Pro Glu Lys Gly Lys Asn Gly Ile 435
440 445 Ala Ile Asn Thr Met Asp Asp Phe
Asn Ser Arg Pro Ile Pro Leu Asn 450 455
460 Lys Glu Gln Leu Asn Thr Tyr Leu Ser Asn Lys Lys Pro
Ile Leu Leu465 470 475
480 Glu Thr Asp Gln Val Glu Gly Lys Tyr Ala Ile Lys Asp Thr Asn Gly
485 490 495 Asn Ile Thr Ile
Ala Gly Asp Trp Asn Gly Ile Thr Asp Glu Ile Ser 500
505 510 Ala Lys Thr Ala Ser Ile Ile Val Asp
Asn Gly Asn Gln Met Ser Glu 515 520
525 Lys Arg Val Ala Ala Lys Asp Tyr Thr Asn Pro Glu Asp Lys
Thr Pro 530 535 540
Asn Leu Ser Val Lys Glu Ala Leu Lys Leu Ala Tyr Pro Asp Glu Ile545
550 555 560 Glu Glu Lys Asp Gly
Leu Leu Phe Tyr Asn Asp Gln Pro Ile Phe Glu 565
570 575 Ala Ser Val Gln Ser Tyr Val Asp Glu Tyr
Thr Ala Lys Gln Ile Arg 580 585
590 Lys Gln Leu Asn Asp Ser Thr Gly Ser Phe Lys Asp Val Lys Asn
Leu 595 600 605 Tyr
Asp Val Lys Leu Glu Pro Lys Met Asn Phe Thr Ile Lys Thr Ser 610
615 620 Thr Leu Tyr Asp Gly Gly
Glu Ser Asp Asn Thr Lys Ile Gly Asn Trp625 630
635 640 Tyr Tyr Thr Tyr Val Val Asn Gly Gly Asn Thr
Gly Lys Lys Gln Tyr 645 650
655 Arg Ser Ala Asn Lys Gly Ala Phe Thr Glu Leu Ser Thr Glu Ser Lys
660 665 670 Asn Lys Leu
Lys Lys Asn Ile Asp Tyr Tyr Val Ser Leu Tyr Met Lys 675
680 685 Ala Asp Ser Lys Val Ser Val Asp
Ile Glu Ile Asp Gly Lys Gln Glu 690 695
700 Ser Ile Val Thr Asp Asn Ile Thr Leu Asp His Val Gly
Tyr Gln Arg705 710 715
720 Ile Asn Ile Leu Val Pro Asn Leu Glu Gly Asn Glu Ile Asn Thr Ile
725 730 735 Ser Ile Lys Gly
Asp Gly Gln Thr Asn Val Tyr Trp Asp Asp Val Ser 740
745 750 Phe Val Glu Val Gly Ala Glu Glu Ile
Glu Tyr Lys Asp Pro Val Pro 755 760
765 Gln Phe Asp Ile Ile Glu Gly Asp Phe Asp Phe Phe Gly Asp
Pro Leu 770 775 780
Ala Val Lys Tyr His Asp Ala Thr Tyr Phe Ile Asp Ser Pro Leu Ile785
790 795 800 Thr Gln Thr Pro Gly
Thr Phe Ser Phe Thr Tyr Lys Val Ile Gly Glu 805
810 815 Gln Thr Lys Thr Val Leu Asp Ser Gly Ser
Gly Lys Asn Ala Asn Arg 820 825
830 Ile Asn Leu Asp Phe Lys Asn Val Lys Ser Asp Arg Ser Phe Leu
Tyr 835 840 845 Thr
Leu Ser Cys Lys Asp Asp Leu Trp Gly Ser Thr Arg Thr Ala Val 850
855 860 Val Arg Ile Phe Ala Val
Asp865 870 51785PRTBrevibacillus laterosporus 51Met
Thr Tyr Met Lys Lys Lys Leu Val Ser Val Val Thr Cys Ala Leu1
5 10 15 Leu Ala Pro Met Phe Leu
Asn Gly Asn Val Asn Pro Val Tyr Ala Asp 20 25
30 Asn Gln Thr Asn Gln Leu Ser Thr Ala Gln Glu
Asn Gln Glu Lys Glu 35 40 45
Val Asp Arg Lys Gly Leu Leu Gly Tyr Tyr Phe Lys Gly Lys Glu Phe
50 55 60 Asn His Leu
Thr Leu Phe Ala Pro Thr Arg Asp Asn Thr Leu Ile Tyr65 70
75 80 Asp Gln Gln Thr Val Asp Phe Leu
Leu Asp Lys Lys Gln Gln Glu Tyr 85 90
95 Gln Ser Ile Arg Trp Ile Gly Leu Ile Gln Ser Lys Glu
Thr Gly Asp 100 105 110
Phe Thr Phe Asn Leu Ser Asp Asp Lys Asn Ala Ile Ile Glu Ile Asp
115 120 125 Gly Lys Thr Ile
Ser His Lys Gly Gln Asn Lys Gln Val Val His Leu 130
135 140 Glu Lys Gly Lys Leu Val Pro Ile
Lys Ile Glu Tyr Gln Ser Asp Gln145 150
155 160 Ile Val Asn Arg Asp Ser Lys Ile Phe Lys Glu Phe
Lys Leu Phe Lys 165 170
175 Val Asp Ser Gln Gln Gln Ser His Gln Val Gln Leu Asp Glu Leu Arg
180 185 190 Asn Pro Glu
Phe Asn Lys Lys Glu Thr Gln Gln Phe Leu Glu Lys Ala 195
200 205 Ser Lys Thr Asn Ile Phe Thr Gln
Asn Met Lys Arg Asp Glu Asp Ala 210 215
220 Thr Asp Thr Asp Gly Asp Ser Ile Pro Asp Leu Trp Glu
Glu Asn Gly225 230 235
240 Tyr Thr Ile Gln Asn Lys Val Ala Val Lys Trp Asp Asp Ser Phe Ala
245 250 255 Ala Lys Gly Tyr
Thr Lys Phe Val Ser Asn Pro Phe Asp Ser His Thr 260
265 270 Val Gly Asp Pro Tyr Thr Asp Tyr Glu
Lys Ala Ala Arg Asp Leu Asp 275 280
285 Leu Ala Asn Ala Lys Glu Thr Phe Asn Pro Leu Val Ala Ala
Phe Pro 290 295 300
Ser Val Asn Val Asn Leu Glu Lys Val Ile Leu Ser Pro Asn Glu Asp305
310 315 320 Leu Ser Asn Ser Val
Glu Ser His Ser Ser Thr Asn Trp Ser Tyr Thr 325
330 335 Asn Thr Glu Gly Ala Ser Ile Glu Ala Gly
Ser Gly Pro Leu Gly Ile 340 345
350 Ser Tyr Gly Val Ser Ala Asn Tyr Gln His Ser Glu Thr Val Ala
Lys 355 360 365 Glu
Trp Gly Thr Ser Thr Gly Asn Thr Ser Gln Phe Asn Thr Ala Ser 370
375 380 Ala Gly Tyr Leu Asn Ala
Asn Val Arg Tyr Asn Asn Val Gly Thr Gly385 390
395 400 Ala Ile Tyr Glu Val Lys Pro Thr Thr Gly Phe
Val Leu Asp Asn Asp 405 410
415 Thr Val Ala Thr Ile Thr Ala Lys Ser Asn Ser Thr Ala Leu Ser Ile
420 425 430 Ser Pro Gly
Glu Ser Tyr Pro Lys Lys Gly Gln Asn Gly Ile Ala Ile 435
440 445 Asn Thr Met Asp Asp Phe Asn Ser
His Pro Ile Thr Leu Asn Lys Gln 450 455
460 Gln Leu Asp Gln Ile Phe Asn Asn Lys Pro Leu Met Leu
Glu Thr Asn465 470 475
480 Gln Ala Asp Gly Val Tyr Lys Ile Lys Asp Thr Ser Gly Asn Ile Val
485 490 495 Thr Gly Gly Glu
Trp Asn Gly Val Ile Gln Gln Ile Gln Ala Lys Thr 500
505 510 Ala Ser Ile Ile Val Asp Thr Gly Glu
Gly Val Ser Glu Lys Arg Val 515 520
525 Ala Ala Lys Asp Tyr Asp Asn Pro Glu Asp Lys Thr Pro Ser
Leu Ser 530 535 540
Leu Lys Glu Ala Leu Lys Leu Gly Tyr Pro Glu Glu Ile Lys Glu Lys545
550 555 560 Asp Gly Leu Leu Tyr
Tyr Asn Asp Lys Pro Ile Tyr Glu Ser Ser Val 565
570 575 Met Thr Tyr Leu Asp Glu Asn Thr Ala Lys
Glu Val Lys Glu Gln Leu 580 585
590 Asn Asp Ile Thr Gly Lys Phe Lys Asp Val Lys Gln Leu Phe Asp
Val 595 600 605 Lys
Leu Thr Pro Lys Met Asn Phe Thr Ile Lys Leu Ala Thr Leu Tyr 610
615 620 Asp Gly Ala Glu Asp Gly
Ser Ser Pro Thr Asp Val Gly Ile Ser Ser625 630
635 640 Pro Leu Gly Glu Trp Ala Phe Lys Pro Asp Ile
Asn Asn Val Glu Gly 645 650
655 Gly Asn Thr Gly Lys Arg Gln Tyr Gln Leu Ser Lys Asn Lys Asp Gly
660 665 670 Tyr Tyr Tyr
Gly Met Leu Ala Leu Ser Pro Glu Val Ser Asn Lys Leu 675
680 685 Lys Lys Asn Tyr Gln Tyr Tyr Ile
Ser Met Ser Ile Lys Ala Asp Ala 690 695
700 Gly Val Glu Pro Thr Val Thr Val Met Asp Asn Leu Leu
Asn Gly Ile705 710 715
720 Val Asp Lys Lys Leu Lys Leu Ser Ser Asn Gly Tyr Gln Arg Phe Asp
725 730 735 Ile Leu Val Asp
Asn Ser Glu Ser His Pro Ile Asn Val Met Val Ile 740
745 750 Asp Leu Gly Val Ser Ser Gln Asp Tyr
Asn Asn Tyr Ser Lys Asn Ile 755 760
765 Tyr Ile Asp Asp Ile Thr Ile Thr Glu Val Ser Ala Met Lys
Val Lys 770 775 780
Asn785 52462PRTBacillus cereus 52Met Lys Arg Met Glu Gly Lys Leu Phe Met
Val Ser Lys Lys Leu Gln1 5 10
15 Val Val Thr Lys Thr Val Leu Leu Ser Thr Val Phe Ser Ile Ser
Leu 20 25 30 Leu
Asn Asn Glu Val Ile Lys Ala Glu Gln Leu Asn Ile Asn Ser Gln 35
40 45 Ser Lys Tyr Thr Asn Leu
Gln Asn Leu Lys Ile Thr Asp Lys Val Glu 50 55
60 Asp Phe Lys Glu Asp Lys Glu Lys Ala Lys Glu
Trp Gly Lys Glu Lys65 70 75
80 Glu Lys Glu Trp Lys Leu Thr Ala Thr Glu Lys Gly Lys Met Asn Asn
85 90 95 Phe Leu Asp
Asn Lys Asn Asp Ile Lys Thr Asn Tyr Lys Glu Ile Thr 100
105 110 Phe Ser Met Ala Gly Ser Phe Glu
Asp Glu Ile Lys Asp Leu Lys Glu 115 120
125 Ile Asp Lys Met Phe Asp Lys Thr Asn Leu Ser Asn Ser
Ile Ile Thr 130 135 140
Tyr Lys Asn Val Glu Pro Thr Thr Ile Gly Phe Asn Lys Ser Leu Thr145
150 155 160 Glu Gly Asn Thr Ile
Asn Ser Asp Ala Met Ala Gln Phe Lys Glu Gln 165
170 175 Phe Leu Asp Arg Asp Ile Lys Phe Asp Ser
Tyr Leu Asp Thr His Leu 180 185
190 Thr Ala Gln Gln Val Ser Ser Lys Glu Arg Val Ile Leu Lys Val
Thr 195 200 205 Val
Pro Ser Gly Lys Gly Ser Thr Thr Pro Thr Lys Ala Gly Val Ile 210
215 220 Leu Asn Asn Ser Glu Tyr
Lys Met Leu Ile Asp Asn Gly Tyr Met Val225 230
235 240 His Val Asp Lys Val Ser Lys Val Val Lys Lys
Gly Val Glu Cys Leu 245 250
255 Gln Ile Glu Gly Thr Leu Lys Lys Ser Leu Asp Phe Lys Asn Asp Ile
260 265 270 Asn Ala Glu
Ala His Ser Trp Gly Met Lys Asn Tyr Glu Glu Trp Ala 275
280 285 Lys Asp Leu Thr Asp Ser Gln Arg
Glu Ala Leu Asp Gly Tyr Ala Arg 290 295
300 Gln Asp Tyr Lys Glu Ile Asn Asn Tyr Leu Arg Asn Gln
Gly Gly Ser305 310 315
320 Gly Asn Glu Lys Leu Asp Ala Gln Ile Lys Asn Ile Ser Asp Ala Leu
325 330 335 Gly Lys Lys Pro
Ile Pro Glu Asn Ile Thr Val Tyr Arg Trp Cys Gly 340
345 350 Met Pro Glu Phe Gly Tyr Gln Ile Ser
Asp Pro Leu Pro Ser Leu Lys 355 360
365 Asp Phe Glu Glu Gln Phe Leu Asn Thr Ile Lys Glu Asp Lys
Gly Tyr 370 375 380
Met Ser Thr Ser Leu Ser Ser Glu Arg Leu Ala Ala Phe Gly Ser Arg385
390 395 400 Lys Ile Ile Leu Arg
Leu Gln Val Pro Lys Gly Ser Thr Gly Ala Tyr 405
410 415 Leu Ser Ala Ile Gly Gly Phe Ala Ser Glu
Lys Glu Ile Leu Leu Asp 420 425
430 Lys Asp Ser Lys Tyr His Ile Asp Lys Val Thr Glu Val Ile Ile
Lys 435 440 445 Gly
Val Lys Arg Tyr Val Val Asp Ala Thr Leu Leu Thr Asn 450
455 460 53775PRTBacillus thuringiensis 53Met Lys
Asn Met Lys Lys Lys Leu Ala Ser Val Val Thr Cys Thr Leu1 5
10 15 Leu Ala Pro Met Phe Leu Asn
Gly Asn Val Asn Ala Val Tyr Ala Asp 20 25
30 Ser Lys Thr Asn Gln Ile Ser Thr Thr Gln Lys Asn
Gln Gln Lys Glu 35 40 45
Met Asp Arg Lys Gly Leu Leu Gly Tyr Tyr Phe Lys Gly Lys Asp Phe
50 55 60 Ser Asn Leu
Thr Met Phe Ala Pro Thr Arg Asp Asn Thr Leu Ile Tyr65 70
75 80 Asp Gln Gln Thr Ala Asn Lys Leu
Leu Asp Lys Lys Gln Gln Gln Tyr 85 90
95 Gln Ser Ile Arg Trp Ile Gly Leu Ile Gln Ser Lys Glu
Lys Gly Asp 100 105 110
Phe Thr Phe Asn Leu Ser Glu Asp Glu Gln Ala Ile Ile Glu Ile Asp
115 120 125 Gly Lys Ile Ile
Ser Asn Lys Gly Lys Glu Lys Gln Val Val His Leu 130
135 140 Glu Lys Glu Lys Leu Val Pro Ile
Lys Ile Glu Tyr Gln Ser Asp Thr145 150
155 160 Lys Phe Asn Ile Asp Ser Lys Thr Phe Lys Glu Phe
Lys Leu Phe Lys 165 170
175 Ile Asp Ser Gln Asn Gln Ser Gln Gln Val Lys Arg Asp Glu Leu Arg
180 185 190 Asn Pro Glu
Phe Asn Lys Lys Glu Ser Arg Glu Phe Leu Ala Lys Ala 195
200 205 Ser Lys Thr Asn Phe Phe Met Gln
Lys Met Lys Arg Asp Ile Asp Glu 210 215
220 Asp Thr Asp Thr Asp Gly Asp Ser Ile Pro Asp Leu Trp
Glu Glu Asn225 230 235
240 Gly Tyr Thr Ile Gln Asn Lys Val Ala Val Lys Trp Asp Asp Lys Phe
245 250 255 Ala Gln Gln Gly
Tyr Val Lys Tyr Leu Ser Ser Pro Tyr Gln Ala His 260
265 270 Thr Val Gly Asp Pro Tyr Thr Asp Trp
Glu Lys Ala Ala Gly Asp Ile 275 280
285 Pro Lys Ser Asn Ala Ala Ala Thr Arg Asn Pro Leu Val Ala
Ala Phe 290 295 300
Pro Ser Ile Asn Val Asp Met Arg Lys Met Ile Leu Ser Lys Asp Ser305
310 315 320 Asn Leu Ser Asn Ser
Ala Glu Ala His Ser Asn Asn Ser Tyr Thr Tyr 325
330 335 Ala Asn Ser Glu Gly Ala Ser Ile Glu Ala
Gly Phe Gly Pro Lys Gly 340 345
350 Phe Ser Phe Gly Val Ser Ala Asn Tyr Gln His Thr Glu Thr Val
Gly 355 360 365 Ser
Asp Trp Gly Asn Ser Lys Ser Asn Thr Glu Gln Phe Asn Ser Ala 370
375 380 Ser Ala Gly Tyr Leu Asn
Ala Asn Val His Tyr Asn Asn Val Gly Thr385 390
395 400 Gly Gly Ile Tyr Asp Ala Gln Pro Thr Thr Ser
Phe Ile Leu Gln Asp 405 410
415 Ser Thr Ile Ala Thr Ile Thr Ala Lys Ser Asn Ala Thr Ala Leu Ser
420 425 430 Ile Pro Ser
Gly Asp Arg Tyr Pro Ala Ser Lys Glu Gly Ile Ser Leu 435
440 445 Lys Thr Met Asp Asp Phe Asn Ser
His Pro Ile Thr Leu Asn Lys Pro 450 455
460 Gln Leu Asp Ala Val Leu Asn Asn Glu Val Ile Lys Ile
Asn Thr Asp465 470 475
480 Gln Thr Asp Gly Arg Tyr Gly Ile Ile Gly Val Asp Gly Lys Ala Glu
485 490 495 Ile Gly Asp Arg
Trp Ser Pro Ile Ile Asp Glu Ile Lys Gly Arg Thr 500
505 510 Ala Ser Ile Ile Ile Asp Pro Ala Asp
Gly Lys Ala Leu Glu Thr Arg 515 520
525 Ile Ala Ala Lys Asp Tyr Lys Asn Pro Glu Asp Lys Thr Pro
Ser Leu 530 535 540
Thr Ile Lys Glu Gly Leu Lys Ile Ala Tyr Pro Glu Ser Ile Ser Glu545
550 555 560 Asp Lys Asp Gly Ile
Leu Phe Tyr Glu Tyr Lys Asn Asp Glu Gly Lys 565
570 575 Val Thr Lys Lys Gln Leu Ser Glu Glu Asn
Ile Met Pro Tyr Leu Asp 580 585
590 Glu Asp Thr Ser Lys Glu Phe Glu Arg Gln Leu Ser Asp Gly Ser
Ala 595 600 605 Lys
Gly Leu Tyr Asp Ile Lys Leu Thr Pro Lys Met Asn Ile Thr Ile 610
615 620 Arg Leu Ala Thr Val Thr
Leu Gly Phe Asp Asp Gln Phe Ser Ala Tyr625 630
635 640 Pro Trp Glu Asn Ala Thr Trp Ser Asp Lys Phe
Gly Asn Leu Arg Leu 645 650
655 Gly Ser Leu Ala Ile Pro Gln Glu Ser Lys Tyr Thr Ile Pro Lys Asp
660 665 670 Lys Val Lys
Pro Asn Tyr Asp Tyr Leu Ile Thr Gly Tyr Ile Lys His 675
680 685 Asp Phe Thr Thr Asp Asn Glu Ser
Leu Gly Ile Val Ala Phe Thr Lys 690 695
700 Lys Asp Asn Phe Glu Met Trp Asn Met Gly Thr Ser Ile
Phe Ser Gln705 710 715
720 Asn Ser Gly Gly Glu Phe Lys Lys Phe Thr Ile Lys Thr Gln Asn Ile
725 730 735 Ser Gly Asp Tyr
Ile Leu Asp Ser Ile Gln Leu Met Lys Arg Asn Asn 740
745 750 Asp Val Asn Lys Ile Asp Ser Tyr Leu
Asp Asp Ile Ser Ile Ile Pro 755 760
765 Ile Gly Pro Asn Lys Ser Arg 770 775
54126PRTBacillus thuringiensis 54Met Thr Val Tyr Asn Ala Thr Phe Thr Ile
Asn Phe Tyr Asn Glu Gly1 5 10
15 Glu Trp Gly Gly Pro Glu Pro Tyr Gly Tyr Ile Lys Ala Tyr Leu
Thr 20 25 30 Asn
Pro Asp His Asp Phe Glu Ile Trp Lys Gln Asp Asp Trp Gly Lys 35
40 45 Ser Thr Pro Glu Arg Ser
Thr Tyr Thr Gln Thr Ile Lys Ile Ser Ser 50 55
60 Asp Thr Gly Ser Pro Ile Asn Gln Met Cys Phe
Tyr Gly Asp Val Lys65 70 75
80 Glu Tyr Asp Val Gly Asn Ala Asp Asp Ile Leu Ala Tyr Pro Ser Gln
85 90 95 Lys Val Cys
Ser Thr Pro Gly Val Thr Val Arg Leu Asp Gly Asp Glu 100
105 110 Lys Gly Ser Tyr Val Thr Ile Lys
Tyr Ser Leu Thr Pro Ala 115 120
125 55462PRTBacillus cereus 55Met Lys Arg Met Glu Gly Lys Leu Phe Met
Val Ser Lys Lys Leu Gln1 5 10
15 Val Val Thr Lys Thr Val Leu Leu Ser Thr Val Phe Ser Ile Ser
Leu 20 25 30 Leu
Asn Asn Glu Val Ile Lys Ala Glu Gln Leu Asn Ile Asn Ser Gln 35
40 45 Ser Lys Tyr Thr Asn Leu
Gln Asn Leu Lys Ile Thr Asp Lys Val Glu 50 55
60 Asp Phe Lys Glu Asp Lys Glu Lys Ala Lys Glu
Trp Gly Lys Glu Lys65 70 75
80 Glu Lys Glu Trp Lys Leu Thr Ala Thr Glu Lys Gly Lys Met Asn Asn
85 90 95 Phe Leu Asp
Asn Lys Asn Asp Ile Lys Thr Asn Tyr Lys Glu Ile Thr 100
105 110 Phe Ser Met Ala Gly Ser Phe Glu
Asp Glu Ile Lys Asp Leu Lys Glu 115 120
125 Ile Asp Lys Met Phe Asp Lys Thr Asn Leu Ser Asn Ser
Ile Ile Thr 130 135 140
Tyr Lys Asn Val Glu Pro Thr Thr Ile Gly Phe Asn Lys Ser Leu Thr145
150 155 160 Glu Gly Asn Thr Ile
Asn Ser Asp Ala Met Ala Gln Phe Lys Glu Gln 165
170 175 Phe Leu Asp Arg Asp Ile Lys Phe Asp Ser
Tyr Leu Asp Thr His Leu 180 185
190 Thr Ala Gln Gln Val Ser Ser Lys Glu Arg Val Ile Leu Lys Val
Thr 195 200 205 Val
Pro Ser Gly Lys Gly Ser Thr Thr Pro Thr Lys Ala Gly Val Ile 210
215 220 Leu Asn Asn Ser Glu Tyr
Lys Met Leu Ile Asp Asn Gly Tyr Met Val225 230
235 240 His Val Asp Lys Val Ser Lys Val Val Lys Lys
Gly Val Glu Cys Leu 245 250
255 Gln Ile Glu Gly Thr Leu Lys Lys Ser Leu Asp Phe Lys Asn Asp Ile
260 265 270 Asn Ala Glu
Ala His Ser Trp Gly Met Lys Asn Tyr Glu Glu Trp Ala 275
280 285 Lys Asp Leu Thr Asp Ser Gln Arg
Glu Ala Leu Asp Gly Tyr Ala Arg 290 295
300 Gln Asp Tyr Lys Glu Ile Asn Asn Tyr Leu Arg Asn Gln
Gly Gly Ser305 310 315
320 Gly Asn Glu Lys Leu Asp Ala Gln Ile Lys Asn Ile Ser Asp Ala Leu
325 330 335 Gly Lys Lys Pro
Ile Pro Glu Asn Ile Thr Val Tyr Arg Trp Cys Gly 340
345 350 Met Pro Glu Phe Gly Tyr Gln Ile Ser
Asp Pro Leu Pro Ser Leu Lys 355 360
365 Asp Phe Glu Glu Gln Phe Leu Asn Thr Ile Lys Glu Asp Lys
Gly Tyr 370 375 380
Met Ser Thr Ser Leu Ser Ser Glu Arg Leu Ala Ala Phe Gly Ser Arg385
390 395 400 Lys Ile Ile Leu Arg
Leu Gln Val Pro Lys Gly Ser Thr Gly Ala Tyr 405
410 415 Leu Ser Ala Ile Gly Gly Phe Ala Ser Glu
Lys Glu Ile Leu Leu Asp 420 425
430 Lys Asp Ser Lys Tyr His Ile Asp Lys Val Thr Glu Val Ile Ile
Lys 435 440 445 Gly
Val Lys Arg Tyr Val Val Asp Ala Thr Leu Leu Thr Asn 450
455 460 56457PRTBrevibacillus laterosporus 56Met
Ile Val Ile Ile Phe Thr Asn Val Lys Gly Gly Asn Glu Leu Lys1
5 10 15 Lys Asn Phe Tyr Lys Asn
Leu Ile Cys Met Ser Ala Leu Leu Leu Ala 20 25
30 Met Pro Ile Ser Ser Asn Val Thr Tyr Ala Tyr
Gly Ser Glu Lys Val 35 40 45
Asp Tyr Leu Val Lys Thr Thr Asn Asn Thr Glu Asp Phe Lys Glu Asp
50 55 60 Lys Glu Lys
Ala Lys Glu Trp Gly Lys Glu Lys Glu Lys Glu Trp Lys65 70
75 80 Leu Thr Val Thr Glu Lys Thr Arg
Met Asn Asn Phe Leu Asp Asn Lys 85 90
95 Asn Asp Ile Lys Lys Asn Tyr Lys Glu Ile Thr Phe Ser
Met Ala Gly 100 105 110
Ser Phe Glu Asp Glu Ile Lys Asp Leu Lys Glu Ile Asp Lys Met Phe
115 120 125 Asp Lys Ala Asn
Leu Ser Ser Ser Ile Val Thr Tyr Lys Asn Val Glu 130
135 140 Pro Ser Thr Ile Gly Phe Asn Lys
Pro Leu Thr Glu Gly Asn Thr Ile145 150
155 160 Asn Thr Asp Val Gln Ala Gln Phe Lys Glu Gln Phe
Leu Gly Lys Asp 165 170
175 Ile Lys Phe Asp Ser Tyr Leu Asp Thr His Leu Thr Ala Gln Asn Val
180 185 190 Ser Ser Lys
Glu Arg Ile Ile Leu Gln Val Thr Val Pro Ser Gly Lys 195
200 205 Gly Ser Thr Ile Pro Thr Lys Ala
Gly Val Ile Leu Asn Asn Asn Glu 210 215
220 Tyr Lys Met Leu Ile Asp Asn Gly Tyr Val Leu His Val
Asp Asn Ile225 230 235
240 Ser Lys Val Val Lys Lys Gly Tyr Glu Cys Leu Gln Ile Gln Gly Thr
245 250 255 Leu Lys Lys Ser
Leu Asp Phe Lys Asn Asp Ile Asn Ala Glu Ala His 260
265 270 Arg Trp Gly Met Lys Asn Tyr Glu Gly
Trp Ala Lys Asn Leu Thr Asp 275 280
285 Pro Gln Arg Glu Ala Leu Asp Gly Tyr Ala Arg Gln Asp Tyr
Lys Gln 290 295 300
Ile Asn Asp Tyr Leu Arg Asn Gln Gly Gly Ser Gly Asn Glu Lys Leu305
310 315 320 Asp Thr Gln Ile Lys
Asn Ile Ser Glu Ala Leu Glu Lys Gln Pro Ile 325
330 335 Pro Glu Asn Ile Thr Val Tyr Arg Trp Cys
Gly Met Ala Glu Phe Gly 340 345
350 Tyr Gln Ile Ser Asp Pro Leu Pro Ser Leu Lys Glu Met Glu Glu
Lys 355 360 365 Phe
Leu Asn Thr Met Lys Glu Asp Lys Gly Tyr Met Ser Thr Ser Leu 370
375 380 Ser Ser Glu Arg Leu Ser
Ala Phe Gly Ser Arg Lys Phe Ile Leu Arg385 390
395 400 Leu Gln Val Pro Lys Gly Ser Thr Gly Ala Tyr
Leu Ser Ala Ile Gly 405 410
415 Gly Phe Ala Ser Glu Lys Glu Ile Leu Ile Asp Lys Asp Ser Asn Tyr
420 425 430 His Ile Asp
Lys Ile Thr Glu Val Val Ile Lys Gly Val Lys Arg Tyr 435
440 445 Val Val Asp Ala Thr Leu Leu Thr
Lys 450 455 57454PRTClostridium perfringens
57Met Lys Lys Val Asn Lys Ser Ile Ser Val Phe Leu Ile Leu Tyr Leu1
5 10 15 Ile Leu Thr Ser
Ser Phe Pro Ser Tyr Thr Tyr Ala Gln Asp Leu Gln 20
25 30 Ile Ala Ser Asn Tyr Ile Thr Asp Arg
Ala Phe Ile Glu Arg Pro Glu 35 40
45 Asp Phe Leu Lys Asp Lys Glu Asn Ala Ile Gln Trp Glu Lys
Lys Glu 50 55 60
Ala Glu Arg Val Glu Lys Asn Leu Asp Thr Leu Glu Lys Glu Ala Leu65
70 75 80 Glu Leu Tyr Lys Lys
Asp Ser Glu Gln Ile Ser Asn Tyr Ser Gln Thr 85
90 95 Arg Gln Tyr Phe Tyr Asp Tyr Gln Ile Glu
Ser Asn Pro Arg Glu Lys 100 105
110 Glu Tyr Lys Asn Leu Arg Asn Ala Ile Ser Lys Asn Lys Ile Asp
Lys 115 120 125 Pro
Ile Asn Val Tyr Tyr Phe Glu Ser Pro Glu Lys Phe Ala Phe Asn 130
135 140 Lys Glu Ile Arg Thr Glu
Asn Gln Asn Glu Ile Ser Leu Glu Lys Phe145 150
155 160 Asn Glu Leu Lys Glu Thr Ile Gln Asp Lys Leu
Phe Lys Gln Asp Gly 165 170
175 Phe Lys Asp Val Ser Leu Tyr Glu Pro Gly Asn Gly Asp Glu Lys Pro
180 185 190 Thr Pro Leu
Leu Ile His Leu Lys Leu Pro Lys Asn Thr Gly Met Leu 195
200 205 Pro Tyr Ile Asn Ser Asn Asp Val
Lys Thr Leu Ile Glu Gln Asp Tyr 210 215
220 Ser Ile Lys Ile Asp Lys Ile Val Arg Ile Val Ile Glu
Gly Lys Gln225 230 235
240 Tyr Ile Lys Ala Glu Ala Ser Ile Val Asn Ser Leu Asp Phe Lys Asp
245 250 255 Asp Val Ser Lys
Gly Asp Leu Trp Gly Lys Glu Asn Tyr Ser Asp Trp 260
265 270 Ser Asn Lys Leu Thr Pro Asn Glu Leu
Ala Asp Val Asn Asp Tyr Met 275 280
285 Arg Gly Gly Tyr Thr Ala Ile Asn Asn Tyr Leu Ile Ser Asn
Gly Pro 290 295 300
Leu Asn Asn Pro Asn Pro Glu Leu Asp Ser Lys Val Asn Asn Ile Glu305
310 315 320 Asn Ala Leu Lys Leu
Thr Pro Ile Pro Ser Asn Leu Ile Val Tyr Arg 325
330 335 Arg Ser Gly Pro Gln Glu Phe Gly Leu Thr
Leu Thr Ser Pro Glu Tyr 340 345
350 Asp Phe Asn Lys Ile Glu Asn Ile Asp Ala Phe Lys Glu Lys Trp
Glu 355 360 365 Gly
Lys Val Ile Thr Tyr Pro Asn Phe Ile Ser Thr Ser Ile Gly Ser 370
375 380 Val Asn Met Ser Ala Phe
Ala Lys Arg Lys Ile Ile Leu Arg Ile Asn385 390
395 400 Ile Pro Lys Asp Ser Pro Gly Ala Tyr Leu Ser
Ala Ile Pro Gly Tyr 405 410
415 Ala Gly Glu Tyr Glu Val Leu Leu Asn His Gly Ser Lys Phe Lys Ile
420 425 430 Asn Lys Val
Asp Ser Tyr Lys Asp Gly Thr Val Thr Lys Leu Ile Leu 435
440 445 Asp Ala Thr Leu Ile Asn 450
584PRTArtificial Sequenceendoplasmic reticulum retention
sequence 58Lys Asp Glu Leu1 5912DNAArtificial
Sequencenucleotide sequence upstream of the ATG start site of
axmi-011 59gtgatgaaaa aa
1260969DNABacillus thuringiensisCDS(1)...(969) 60gtg atg aaa aaa
atg aat aaa aaa cct atg gta gcg ttg ata tta gcc 48Met Met Lys Lys
Met Asn Lys Lys Pro Met Val Ala Leu Ile Leu Ala1 5
10 15act tcg att ggt ata cct tgt aca ttt aca
cct gga agt gca tta gca 96Thr Ser Ile Gly Ile Pro Cys Thr Phe Thr
Pro Gly Ser Ala Leu Ala 20 25
30gca gaa aat att cag act agt gtt aat gaa aat gta aaa gtt ggt att
144Ala Glu Asn Ile Gln Thr Ser Val Asn Glu Asn Val Lys Val Gly Ile
35 40 45aca gat gtt caa tct gaa ttg aat
aag ata gga gac tat tat tat agt 192Thr Asp Val Gln Ser Glu Leu Asn
Lys Ile Gly Asp Tyr Tyr Tyr Ser 50 55
60aat aac tta gca aat acg act ata aaa cct cct cat cat tgg gat tat
240Asn Asn Leu Ala Asn Thr Thr Ile Lys Pro Pro His His Trp Asp Tyr65
70 75 80aca ctt aaa aaa aat
cct gat aaa gtt gga aca aat ttg gat ttt agt 288Thr Leu Lys Lys Asn
Pro Asp Lys Val Gly Thr Asn Leu Asp Phe Ser 85
90 95att act ggt act gct agt aaa cta aat tat gat
agt gta act cct ata 336Ile Thr Gly Thr Ala Ser Lys Leu Asn Tyr Asp
Ser Val Thr Pro Ile 100 105
110tac att ggg cat aat gaa ttt aat aat gat tca gat cag cct caa aaa
384Tyr Ile Gly His Asn Glu Phe Asn Asn Asp Ser Asp Gln Pro Gln Lys
115 120 125ttt aca act tct aaa ttt act
aaa gct gta aca gag gga aca aca agt 432Phe Thr Thr Ser Lys Phe Thr
Lys Ala Val Thr Glu Gly Thr Thr Ser 130 135
140acc gta aca aat gga ttt aga tta gga aat cca ggt tta aac tta ttt
480Thr Val Thr Asn Gly Phe Arg Leu Gly Asn Pro Gly Leu Asn Leu Phe145
150 155 160act att cca tta
att tta agt gat ggt atg aaa att aat gcg gaa ttt 528Thr Ile Pro Leu
Ile Leu Ser Asp Gly Met Lys Ile Asn Ala Glu Phe 165
170 175aac tct tct act tca gaa tct caa caa aaa
tcg gaa aca aaa aca ata 576Asn Ser Ser Thr Ser Glu Ser Gln Gln Lys
Ser Glu Thr Lys Thr Ile 180 185
190gaa gca tca cct caa aac ata gaa gtt cca gca cat aaa aaa tat aaa
624Glu Ala Ser Pro Gln Asn Ile Glu Val Pro Ala His Lys Lys Tyr Lys
195 200 205gta gat gtt gta ttg gaa caa
aca agc tat tgg gca gat gtt aca ttt 672Val Asp Val Val Leu Glu Gln
Thr Ser Tyr Trp Ala Asp Val Thr Phe 210 215
220aca ggt gaa gga att aat ctt aat act act ata aat gca act gga ata
720Thr Gly Glu Gly Ile Asn Leu Asn Thr Thr Ile Asn Ala Thr Gly Ile225
230 235 240cat act ggg cat
atg gga atg cag gag tca aga aaa ttt tct tgg aac 768His Thr Gly His
Met Gly Met Gln Glu Ser Arg Lys Phe Ser Trp Asn 245
250 255aaa aat acc att gaa tta ttt aat gga cta
aaa caa gag caa aaa aat 816Lys Asn Thr Ile Glu Leu Phe Asn Gly Leu
Lys Gln Glu Gln Lys Asn 260 265
270aat ata cat ggg att aaa ttt agt aat ggg aaa atg aat gca aac gga
864Asn Ile His Gly Ile Lys Phe Ser Asn Gly Lys Met Asn Ala Asn Gly
275 280 285aca ggt aaa gtt gaa ggt att
ttt ggt agt aat cta gtt gta aag gta 912Thr Gly Lys Val Glu Gly Ile
Phe Gly Ser Asn Leu Val Val Lys Val 290 295
300aat gat gtt aca gat cca tta aat cct atc cta gta atg act aaa agt
960Asn Asp Val Thr Asp Pro Leu Asn Pro Ile Leu Val Met Thr Lys Ser305
310 315 320tta aaa taa
969Leu Lys
61322PRTBacillus thuringiensis 61Met Met Lys Lys Met Asn Lys Lys Pro Met
Val Ala Leu Ile Leu Ala1 5 10
15 Thr Ser Ile Gly Ile Pro Cys Thr Phe Thr Pro Gly Ser Ala Leu
Ala 20 25 30 Ala
Glu Asn Ile Gln Thr Ser Val Asn Glu Asn Val Lys Val Gly Ile 35
40 45 Thr Asp Val Gln Ser Glu
Leu Asn Lys Ile Gly Asp Tyr Tyr Tyr Ser 50 55
60 Asn Asn Leu Ala Asn Thr Thr Ile Lys Pro Pro
His His Trp Asp Tyr65 70 75
80 Thr Leu Lys Lys Asn Pro Asp Lys Val Gly Thr Asn Leu Asp Phe Ser
85 90 95 Ile Thr Gly
Thr Ala Ser Lys Leu Asn Tyr Asp Ser Val Thr Pro Ile 100
105 110 Tyr Ile Gly His Asn Glu Phe Asn
Asn Asp Ser Asp Gln Pro Gln Lys 115 120
125 Phe Thr Thr Ser Lys Phe Thr Lys Ala Val Thr Glu Gly
Thr Thr Ser 130 135 140
Thr Val Thr Asn Gly Phe Arg Leu Gly Asn Pro Gly Leu Asn Leu Phe145
150 155 160 Thr Ile Pro Leu Ile
Leu Ser Asp Gly Met Lys Ile Asn Ala Glu Phe 165
170 175 Asn Ser Ser Thr Ser Glu Ser Gln Gln Lys
Ser Glu Thr Lys Thr Ile 180 185
190 Glu Ala Ser Pro Gln Asn Ile Glu Val Pro Ala His Lys Lys Tyr
Lys 195 200 205 Val
Asp Val Val Leu Glu Gln Thr Ser Tyr Trp Ala Asp Val Thr Phe 210
215 220 Thr Gly Glu Gly Ile Asn
Leu Asn Thr Thr Ile Asn Ala Thr Gly Ile225 230
235 240 His Thr Gly His Met Gly Met Gln Glu Ser Arg
Lys Phe Ser Trp Asn 245 250
255 Lys Asn Thr Ile Glu Leu Phe Asn Gly Leu Lys Gln Glu Gln Lys Asn
260 265 270 Asn Ile His
Gly Ile Lys Phe Ser Asn Gly Lys Met Asn Ala Asn Gly 275
280 285 Thr Gly Lys Val Glu Gly Ile Phe
Gly Ser Asn Leu Val Val Lys Val 290 295
300 Asn Asp Val Thr Asp Pro Leu Asn Pro Ile Leu Val Met
Thr Lys Ser305 310 315
320 Leu Lys626DNAArtificial Sequenceribosomal binding sequence 62gtgatg
6635DNAArtificial Sequenceribosomal binding sequence 63ggaga
5
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