Patent application title: POLYPEPTIDE HAVING LARVAE GROWTH INHIBITING OR INSECTICIDAL EFFECT ON SCARABAEIDAE INSECTS AND POLYNUCLEOTIDE ENCODING THE SAME
Inventors:
Masao Tanaka (Chiba, JP)
Tomoko Yokoyama (Chiba, JP)
Moriichi Aoyagi (Katori-Gun, JP)
Makoto Hasegawa (Chiba, JP)
Gaku Ehara (Sakura, JP)
Masaharu Kimura (Ichihara, JP)
Hideji Nishihashi (Sakura, JP)
Assignees:
DAINIPPON INK & CHEMICALS, INC.
IPC8 Class: AA01N3718FI
USPC Class:
514 2
Class name: Drug, bio-affecting and body treating compositions designated organic active ingredient containing (doai) peptide containing (e.g., protein, peptones, fibrinogen, etc.) doai
Publication date: 2010-01-14
Patent application number: 20100009897
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Patent application title: POLYPEPTIDE HAVING LARVAE GROWTH INHIBITING OR INSECTICIDAL EFFECT ON SCARABAEIDAE INSECTS AND POLYNUCLEOTIDE ENCODING THE SAME
Inventors:
Masao Tanaka
Tomoko Yokoyama
Moriichi Aoyagi
Makoto Hasegawa
Gaku Ehara
Masaharu Kimura
Hideji Nishihashi
Agents:
EDWARDS ANGELL PALMER & DODGE LLP
Assignees:
DAINIPPON INK & CHEMICALS, INC.
Origin: BOSTON, MA US
IPC8 Class: AA01N3718FI
USPC Class:
514 2
Patent application number: 20100009897
Abstract:
Through screening using an antibody to a polypeptide that constitutes a
parasporal body in sporangia of Bacillus popilliae and has a larvae
growth inhibiting or insecticidal effect on a Scarabaeidae insect and
through polymerase chain reaction, polynucleotides encoding the
polypeptide and polypeptides similar thereto are isolated.Claims:
1-27. (canceled)
28. A method for controlling a Scarabaeidae insect, which comprises a step of administering orally a Scarabaeidae insect controlling agent comprising as an active ingredient a polypeptide having the amino acid sequence of SEQ ID NO: 20 to the Scarabaeidae insect.
29. The method according to claim 28, wherein the Scarabaeidae insect controlling agent further comprises a component selected from the group consisting of excipients, carriers, nutritive agents and stabilizers.
30. The method according to claim 28, wherein the Scarabaeidae insect controlling agent further comprises sporangia of a bacterium belonging to Bacillus popilliae.
31. The method according to claim 29, wherein the Scarabaeidae insect controlling agent further comprises sporangia of a bacterium belonging to Bacillus popilliae.
32. The method according to claim 29, wherein the Scarabaeidae insect controlling agent further comprises a component selected from the group consisting of mineral fine powders selected from the group consisting of kaolin clay, bentonite, montmorillonite, diatomaceous earth, acid clay, talcs, pearlite, vermiculite, ammonium sulfate, urea, ammonium chloride, ammonium nitrate, wheat bran, chitin, polysaccharides, rice bran, and flour, as solid carriers; fixing agents and dispersants selected from the group consisting of casein, gelatin, gum Arabic, alginic acid, saccharides, synthetic polymers, polyvinyl alcohol, and polyacrylic acids; and cryoprotectants selected from the group consisting of propylene glycol, ethylene glycol, xanthan gum and polyacrylic acids.
33. The method according to claim 29, wherein a ratio of polypeptide to other components is 0.0001 to 100% by mass.
34. The method according to claim 28, wherein the Scarabaeidae insect is selected from the group consisting of Anomala cuprea, Blitopertha orientalis, Popilliae japonica, Phyllopertha diversa, Adoretus tenuimaculatus and Anomala rufocuprea.
35. The method according to claim 28, wherein an application amount of polypeptide is 0.01 to 100 g/m2 for ground dispersion.
Description:
[0001]This application is a divisional application of U.S. application
Ser. No. 11/091,643, filed Mar. 29, 2005 which is a divisional
application of 10/120,544 filed on Apr. 12, 2002 which claims priority on
JP 2001-115754 filed on Apr. 13, 2001 and JP2001-203463 filed on Jul. 4,
2001, which is hereby incorporated by reference herein in its entirety as
if fully set forth herein.
BACKGROUND OF THE INVENTION
[0002]1. Field of the Invention
[0003]The present invention relates to polypeptides having larvae growth inhibiting or insecticidal effect on Scarabaeidae insects, to polynucleotides encoding the same, to controlling agents for controlling Scarabaeidae insects containing the polypeptides as active ingredients, and to a method of controlling Scarabaeidae insects using the polypeptides.
[0004]2. Description of the Related Art
[0005]Larvae of Scarabaeidae insects eat roots of a wide variety of plants such as turf, agricultural crops, horticultural crops and trees and bushes and cause serious damage thereto. A large number of reports have been made on damages caused by, especially, Anomala cuprea, Blitopertha orientalis, and Popillia japonica to a lawn of golf courses, a sweet potato field, and a peanut field. In particular, Anomala cuprea, which is particularly large in size, does considerable harm thereto due to its eager appetite.
[0006]However, conventional ground spreading of chemical pesticides is difficult to give a controlling effect since these larvae inhabit in the ground and are difficult to locate them, and thus it was necessary to spread a large amount of chemical pesticides over a wide range of the surface of the ground and penetrate them into the ground. Therefore, there were concerns about adverse influences of the chemical pesticides on the natural environment and human bodies, and there has been a keen desire for a biological controlling method that can replace the chemical pesticides.
[0007]As one of the biological controlling methods for controlling Scarabaeidae insects, attempts to utilize bacteria belonging to Bacillus popilliae have been tried over a long period of time. The bacteria are parasitic to larvae of Scarabaeidae insects as hosts and infect to them per os usually in a form of sporangia, propagate in large amounts in hemolymph to cause milky disease and finally lead the larvae to death.
[0008]Recently, an amino acid sequence of a polypeptide constituting a parasporal body of a strain belonging to Bacillus popilliae, i.e., Bacillus popilliae subsp. melolonthae H1, and a nucleotide sequence of the gene encoding the polypeptide were partly clarified (J. Bacteriol. Vol. 179, p. 4336-4341 (1997)), and it has been reported that the polypeptide constituting the parasporal body of Bacillus popilliae subsp. melolonthae H1 has a controlling effect on Melolontha melolontha (WO97/14798). However, it was not clarified whether the bacterial strain and a polypeptide thereof have controlling effects on Anomala cuprea, Blitopertha orientalis and Popillia japonica, each of which belongs to different species.
SUMMARY OF THE INVENTION
[0009]An object to be solved according to the present invention is to provide polypeptides having a larvae growth inhibiting effect or an insecticidal effect on Scarabaeidae insects, in particular Anomala cuprea, Blitopertha orientalis and Popillia japonica, transformants having introduced therein a polynucleotide encoding the polypeptide, controlling agents for Scarabaeidae insects containing the polypeptides as active ingredients, controlling agents for Scarabaeidae insects using the polypeptide and sporangia of Bacillus popilliae, and a method of controlling Scarabaeidae insects using the controlling agents.
[0010]The inventors of the present invention have found that the sporangia of certain strains belonging to Bacillus popilliae have a larvae growth inhibiting effect or an insecticidal effect. Subsequently, they have taken out the parasporal body from sporangia of Bacillus popilliae (FIG. 1) containing a spore and a parasporal body, performed screening using an antibody to the polypeptide constituting the parasporal body to thereby isolate a polynucleotide encoding the polypeptide and determined its nucleotide sequence and amino acid sequence. A transformant having introduced therein the polynucleotide by using a vector enabled mass production of the above-mentioned polypeptide. Further, the inventors of the present invention have made it clear that the polypeptide produced by the transformant has a controlling effect on Anomala cuprea and further that use of the polypeptide in combination with the sporangia increases the controlling effect. Thus, the present invention was accomplished.
[0011]Therefore, according to the present invention, there is provided (1) a polypeptide having the amino acid sequence of SEQ ID NO: 2, or the amino acid sequence including substitution, deletion or insertion of one or several numbers of amino acid residues and having a larvae growth inhibiting or insecticidal effect on a Scarabaeidae insect.
[0012]According to the present invention, there is provided (2) a polypeptide having the amino acid sequence of SEQ ID NO: 4, 6, 18 or 20, or the amino acid sequence including substitution, deletion or insertion of one or several numbers of amino acid residues, the polypeptide having a larvae growth inhibiting or insecticidal effect on a Scarabaeidae insect.
[0013]According to the present invention, there is provided (3) a polypeptide having the amino acid sequence of SEQ ID NO: 8 or 10, or the amino acid sequence including substitution, deletion or insertion of one or several numbers of amino acid residues, the polypeptide having a larvae growth inhibiting or insecticidal effect on a Scarabaeidae insect.
[0014]Further, the present invention provides polynucleotides encoding the polypeptides.
[0015]Still further, the present invention provides vectors comprising the polynucleotides and transformants which are introduced the polynucleotides.
[0016]Further, the present invention provides controlling agents for controlling a Scarabaeidae insect containing the polypeptides as active ingredients.
[0017]Further, the present invention provides controlling agents for controlling a Scarabaeidae insect comprising the polypeptides and sporangia of bacteria belonging to Bacillus popilliae as active ingredients.
[0018]Still further, the present invention provides a method of controlling a Scarabaeidae insect, in which the Scarabaeidae insect is subjected to an action of the controlling agent for controlling a Scarabaeidae insect.
[0019]In the present invention, the term "polypeptide" includes peptides consisting of a plurality of amino acids linked through peptide bonds and polypeptides to which other molecules such as glycoproteins are bonded as well as proteins in a form of multimers, etc.
[0020]According to the present invention, polypeptides having a larvae growth inhibiting or an insecticidal effect on Scarabaeidae insects, in particular Anomala cuprea, Blitopertha orientalis and Popilliae japonica, transformants having introduced therein a polynucleotide encoding the polypeptide, a controlling agent for controlling a Scarabaeidae insect comprising the polypeptide as an active ingredient, and a method of controlling a Scarabaeidae insect using the polypeptide and sporangia of Bacillus popilliae as active ingredients.
BRIEF EXPLANATION OF THE DRAWINGS
[0021]FIG. 1 is a schematic diagram of a sporangium containing a spore and a parasporal body;
[0022]FIG. 2 is a diagram illustrating a larvae growth inhibiting effect of a polypeptide produced by the polynucleotide on a Scarabaeidae insect (larva of Anomala cuprea) performed in Example 4;
[0023]FIG. 3 is a diagram illustrating an insecticidal effect of biological tests performed in Example 5;
[0024]FIG. 4 is a diagram illustrating the larvae growth inhibiting effect of the biological tests performed in Example 5;
[0025]FIG. 5 shows alignment of amino acid sequences in Example 11;
[0026]FIG. 6 shows alignment of amino acid sequences in Example 11;
[0027]FIG. 7 shows alignment of amino acid sequences in Example 11;
[0028]FIG. 8 is a diagram illustrating a larvae growth inhibiting effect on a Scarabaeidae insect of a polypeptide produced by a polynucleotide in Example 13 of the present invention (larvae of Anomala cuprea).
DETAILED DESCRIPTION OF THE INVENTION
[0029]Hereinafter, the present invention will be described in detail.
[0030]A parasporal body contained in sporangia of a bacterium belonging to Bacillus popilliae is a proteinaceous aggregate (Hukuhara, Toshihiko, "Insect Pathology", p. 57, 1979), which comprises one kind or plural kinds of polypeptides. The polypeptide of the present invention is one or more polypeptides that constitute the parasporal body in the sporangium of Bacillus popilliae semadara or the like that will be described below. However, the polypeptide of the present invention is different in the amino acid sequence from a polypeptide that constitutes a parasporal body described in WO97/14798, and hence the polypeptide and polynucleotide of the present invention are considered to be novel.
[0031]According to a first aspect of the present invention, there is provided a polypeptide having the amino acid sequence of SEQ ID NO: 2 and having a larvae growth inhibiting effect or an insecticidal effect on a Scarabaeidae insect.
[0032]According to a second aspect of the present invention, there is provided a polypeptide having the amino acid sequence of SEQ ID NO: 4, 6, 18 or 20 and having a larvae growth inhibiting effect or an insecticidal effect on a Scarabaeidae insect.
[0033]According to a third aspect of the present invention, there is provided a polypeptide having the amino acid sequence of SEQ ID NO: 8 or 10 and having a larvae growth inhibiting effect or an insecticidal effect on a Scarabaeidae insect.
[0034]Note that the amino acid sequence of SEQ ID NO: 2 and the nucleotide sequence of SEQ ID NO: 1 are sequences found in the amino acid sequences of SEQ ID NO: 4, 6, 18 or 20 and the nucleotide sequences of SEQ ID NO: 3, 5, 17 or 19 with homology of 95% or more and is considered to be specific to the parasporal body of Bacillus popilliae (cf. Example 11).
[0035]The above-mentioned polypeptide of the present invention may include polypeptides containing substitution, deletion or insertion of one or several numbers of amino acid residues as far as the polypeptides have a larvae growth inhibiting effect or an insecticidal effect on a Scarabaeidae insect. Here, the term "several numbers of" means, specifically, a number of from 2 to 100, preferably from 2 to 50, and more preferably from 2 to 9, although it may vary depending on the position and kinds of amino acid residues in a three-dimensional structure of the polypeptide. These polypeptides including substitution, deletion or insertion of one or several numbers of amino acid residues can be obtained, for example, by introducing a mutation to a polynucleotide encoding the polypeptide by site-specific mutatagenesis and performing transcription and translation of the polynucleotide. These polypeptides of the present invention may be fused polypeptides with other polypeptides. The polypeptides of the present invention may be polypeptides that have the amino acid sequence having homology of 50% or more, preferably 70% or more, and more preferably 90% or more, with the above-mentioned amino acid sequence as far as the polypeptides have a larvae growth inhibiting effect or an insecticidal effect on a Scarabaeidae insect. Further, the polypeptides of the present invention may be polypeptides that have a part of the amino acid sequence of SEQ ID NO: 4, 6, 18 or 20 as far as the polypeptides have a larvae growth inhibiting effect or an insecticidal effect on a Scarabaeidae insect.
[0036]The polynucleotides of the present invention are polynucleotides that encode the above-mentioned polypeptides of the present invention. Codons of each of amino acid residues are not particularly limited as far as the amino acid sequence encoded is the same. Specific examples of the polynucleotide of the present invention include a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 1, a polynucleotide having the nucleotide sequence of the coding region of SEQ ID NO: 3, 5, 17 or 19, and a polynucleotide having the nucleotide sequence of SEQ ID NO: 7 or 9.
[0037]The polynucleotide of the present invention may be a polynucleotide that is hybridizable under the stringent conditions to each of the above-mentioned polynucleotide or a probe that can be prepared from the above-mentioned polynucleotide as far as they encode polypeptides having a larvae growth inhibiting effect or an insecticidal effect on a Scarabaeidae insect. The term "the stringent conditions" used herein refers to a condition under which a so-called specific hybrid is formed and nonspecific hybrid is not formed. Such a condition is a condition in which DNAs having high homology, for example, DNAs having homology of 50% or more, preferably 70% or more, and more preferably 90% or more, are hybridized and DNAs having homology that is lower than the above are not hybridized. Specifically, it includes a condition of 40° C., 1×SSC (0.15 M NaCl, 15 mM Sodium Citrate, pH 7.0) and 0.1% SDS (Sodium Dodecyl Sulfate).
[0038]The polynucleotides of the present invention can be obtained from, for example, bacteria belonging to Bacillus popilliae, more specifically from Bacillus popilliae semadara, FERM P-16818, Bacillus popilliae var. Mame, FERM P-17661, Bacillus popilliae var. popilliae Hime, FERM P-17660, Bacillus popilliae var. popilliae Sakura, FERM P-17662, and Bacillus popilliae Dutky, American Type Culture Collection No. 14706 by the following methods.
[0039]Bacillus popilliae semadara, FERM P-16818 has been deposited by Chiba-ken (1-1 Ichiba-cho, Chuo-ku, Chiba-shi, Chiba-ken, Japan) since May 21, 1998 in National Institute of Bioscience and Human Technology of Agency of Industrial Science and Technology of Ministry of International Trade and Industry (zip code: 305-8566, 1-3 Higashi 1-Chome, Tsukuba-shi, Ibaraki-ken, Japan) (currently, National Institute of Advanced Industrial Science and Technology, International Patent Organism Depositary, Tsukuba Central 6, 1-1, Higashi 1-Chome, Tsukuba-shi, Ibaraki-ken, 305-8566, Japan), and the original deposit was converted to international deposit based on Budapest Treaty on Jun. 10, 2002, and assigned the deposition number of FERM BP-8068.
[0040]Bacillus popilliae var. Mame, FERM P-17661 has been deposited by the applicant of the application since Nov. 25, 1999 in National Institute of Bioscience and Human Technology of Agency of Industrial Science and Technology of Ministry of International Trade and Industry (zip code: 305-8566, 1-3 Higashi 1-Chome, Tsukuba-shi, Ibaraki-ken, Japan) (currently, National Institute of Advanced Industrial Science and Technology, International Patent Organism Depositary, Tsukuba Central 6, 1-1, Higashi 1-Chome, Tsukuba-shi, Ibaraki-ken, 305-8566, Japan), and the original deposit was converted to international deposit based on Budapest Treaty on Jun. 10, 2002, and assigned the deposition number of FERM BP-8069.
[0041]One method of obtaining them includes a screening method by utilizing an antibody. That is, an antibody to a purified parasporal body is prepared in advance. On the other hand, a chromosomal DNA of the above-mentioned bacterial strain is collected, cleaved with an appropriate restriction enzyme and inserted into an expression vector to prepare a chromosomal DNA library. Then, the library is expanded in Escherichia coli or the like and screened by Western blotting utilizing the above-mentioned antibody.
[0042]Another method of obtaining the polynucleotides of the present invention includes a PCR method. An appropriate nucleotide sequence is selected from nucleotide sequences of SEQ ID NOs: 11 to 16, 21 and 22 and nucleotide sequences obtained by adding modifications thereto and partial nucleotide sequences derived from the nucleotide sequences of SEQ ID NOs: 1, 3, 5, 7, 9, 17 and 19, and two kinds of polynucleotides directing opposite to each other are provided. PCR is performed using the polynucleotides as primers and a chromosomal DNA as a template. By cloning an amplified DNA fragment, the polynucleotides of the present invention can be obtained.
[0043]Note that PCR using oligonucleotides having respective nucleotide sequences of SEQ ID NOS: 11 and 13 as primers and a chromosomal DNA of Bacillus popilliae semadara as a template can give rise to polynucleotide amplified fragments of about 4.2 kb having the nucleotide sequences of SEQ ID NOS: 17 and 19, respectively. The nucleotide sequences of these polynucleotides exist adjacent to each other on the chromosomal DNA of Bacillus popilliae semadara and the amino acid sequences (SEQ ID NOS: 18 and 20) of the polypeptides encoded by the polynucleotides have homology of about 73% to each other, which strongly suggests that both the polynucleotides have similar functions.
[0044]Also, the polynucleotides having the nucleotide sequences of SEQ ID NOS: 3, 5, 7, and 9 obtained by PCR using a chromosomal DNA of Bacillus popilliae var. popilliae Mame as a template and polypeptides having amino acid sequences of SEQ ID NOS: 4, 6, 8 and 10, which are translation products thereof, have high homology to the nucleotide sequence of the polynucleotide of Bacillus popilliae semadara and the polypeptide encoded by the polynucleotide (homology of 61 to 88% for the polynucleotide and 59 to 98% for the polypeptide). This suggests that they also have similar functions.
[0045]Further, another method of obtaining the polynucleotides of the present invention includes a hybridization method. After digesting the chromosomal DNA with an appropriate restriction enzyme, a genome library is prepared by utilizing a plasmid vector or a phage vector or the like. On the other hand, appropriate polynucleotide probes having the nucleotide sequences of SEQ ID NOS: 11 to 16, 21 and 22 or nucleotide sequences obtained by adding modifications thereto, or partial nucleotide sequences derived from the nucleotide sequences of SEQ ID NOS: 1, 3, 5, 7, 9, 17 and 19 are provided. By colony hybridization or plaque hybridization in which a library prepared in Escherichia coli or the like by using the vector is screened with a labeled probe, the polynucleotide of the present invention can be obtained.
[0046]The vectors that can be used in the present invention include plasmid vectors, Ti plasmid vectors, phage vectors, phagemid vectors, YAC vectors, virus vectors and the like. The vectors may include, in addition to the polynucleotides of the present invention, sequences that regulate or assist expression of polypeptides (for example, a promoter, an operator, a terminator, an enhancer, and an SD sequence) or marker gene for selecting transformant hosts (for example, an ampicillin resistance gene, a tetracycline resistance gene, a kanamycin resistance gene, and a neomycin resistance gene) and the like. The vectors may comprise plural kinds of polynucleotides of the present invention or a plurality of molecules of the same polynucleotide.
[0047]The hosts that can be used in the present invention include microbes (for example, bacteria such as Escherichia coli, Pseudomonad, and Bacillus, actinomycetes such as Streptomyces, Fungi such as molds and yeasts), plants (for example, turf, sweet potatoes, tobaccos, rice plants, and corn), cultured cells (for example, plant cultured cells such as a tobacco cultured cell, insect cultured cells such as a silkworm moth cultured cell and animal cultured cells such as mouse cultured cell) and the like.
[0048]In the present invention, conventional methods used in genetic engineering can be used in the procedure of constructing a vector and transformation operation of hosts with the vector. For example, the transformation method includes a calcium chloride method, an electroporation method, a PEG method and the like for microbe hosts, an electroporation method, a particle gun method, an agrobacterium method and the like for cultured cells and plant hosts.
[0049]In a case where the host is a microbe or a cultured cell, the method of producing a polypeptide of the present invention can be performed by culturing the microbe or the cultured cell in an appropriate medium and recover the produced polypeptide from the culture. The culture method of culturing a transformed host may be substantially the same as the culture method for the host to be used. Extraction of the polypeptide from the culture and purification thereof, if necessary, may be performed by usually employed methods. On the other hand, in a case where the host is a plant, the transformed plant is grown by the same methods as the methods usually used, and a polypeptide can be extracted from the plant body. From a different standpoint, the transformed plant and its seeds may be provided as is as insect resistant cultigen species. In addition, there is a production method of a polypeptide without using any transformant. It includes, for example, an in vitro cell-free transcription/translation system utilizing a wheat germ extract or an Escherichia coli extract. A polypeptide can be produced by adding a DNA fragment or an RNA fragment, or a vector containing the polynucleotide of the present invention to the transcription/translation system, thereby performing a conventional method for producing a polypeptide.
[0050]The polypeptides of the present invention have a larvae growth inhibiting or insecticidal activity on Scarabaeidae insects and exhibit controlling effects thereon. For this reason, the polypeptides can be applied to turf, agricultural crops or trees as a controlling agent or an active ingredient thereof for the purpose of controlling Scarabaeidae insects. Use of the polypeptides of the present invention in combination with spores or sporangia containing spores and parasporal bodies of bacteria belonging to Bacillus popilliae can further increase the larvae growth inhibiting or insecticidal activity on Scarabaeidae insects.
[0051]The Scarabaeidae insects that are the target of control with the polypeptides of the present invention include Anomala cuprea, Blitopertha orientalis, Popillia japonica, Phyllopertha diversa, Adoretus tenuimaculatus, Anomala rufocuprea and the like. The polypeptides of the present invention are particularly effective on Anomala cuprea and Blitopertha orientalis.
[0052]As the controlling agent of the present invention, the polypeptides produced by the above-mentioned production method may be used as they are in the form of the transformants, as a purified product after isolation or as mixtures of these with substances used for culture/reaction systems of these. Alternatively, they may be used by adding excipients, carriers, nutritive agents, stabilizers and the like known and commonly used in agricultural chemicals to convert them into an optional preparation form such as dust, wettable powder, or a flowable agent.
[0053]The above-mentioned optional component includes: mineral fine powders such as kaolin clay, bentonite, montmorillonite, diatomaceous earth, acid clay, talcs, pearlite, and vermiculite, inorganic salts such as ammonium sulfate, urea, ammonium chloride, and ammonium nitrate, organic fine powders such as wheat bran, chitin, polysaccharides, rice bran, and flour, as solid carriers; fixing agents and dispersants such as casein, gelatin, gum Arabic, alginic acid, saccharides, synthetic polymers (polyvinyl alcohol, polyacrylic acids, etc.) as adjutants; and cryoprotectants such as propylene glycol and ethylene glycol, natural polysaccharides such as xanthan gum, thickeners such as polyacrylic acids as other components.
[0054]The ratio of the polypeptides of the present invention contained in the controlling agent is not particularly limited as far as they exhibit a controlling effect on Scarabaeidae insects, but it is preferably 0.0001 to 100 mass %.
[0055]The method of application of the controlling agent is selected as appropriate depending on the use form such as the preparation form, crops and the like and includes, for example, liquid dispersion on the ground, solid dispersion on the ground, liquid dispersion in the air, solid dispersion in the air, application in installations, application for mixing with soil, application for perfusion into soil, and the like methods.
[0056]Also, it is possible to apply the controlling agent of the present invention after mixing with other agents, i.e., pesticides, nematicides, acaricides, herbicides, microbicides, plant growth regulators, fertilizers, soil conditioning materials (peat, humic acid materials, polyvinyl alcohol-based materials, etc.) and the like, to apply it with alternately applying the other agents without mixing, or to apply it together with the other agents simultaneously.
[0057]The application amount of the controlling agent of the present invention cannot be uniquely defined since it may vary depending on the kind of Scarabaeidae insect, kind of plant to which the agent is applied, preparation form of the agent and the like. For example, the application amount of the polypeptide may be 0.01 to 100 g/m2, preferably about 0.1 to 50 g/m2 for dispersion on the ground.
[0058]Further, it is possible to use mixtures of the controlling agent of the present invention with other biocontrol agents or polypeptides derived from microbes (for example, the parasporal bodies of another strain of Bacillus popilliae or the parasporal bodies of Bacillus thuringiensis) or microbial spores (for example, spores of Bacillus popilliae, Bacillus thuringiensis, or Bacillus subtilis).
BEST MODE FOR CARRYING OUT THE INVENTION
[0059]Hereinafter, the present invention will be illustrated more specifically. However, the scope of the present invention is not limited thereto.
Example 1
[0060]A suspension of sporangia of Bacillus popilliae prepared from larvae of Anomala cuprea infected with milky disease was subjected to treatment by a French press to take out spores and parasporal bodies from the sporangia. Only the parasporal bodies were recovered by a two-phase separation method by using sodium dextran sulfate (produced by Sigma Co., Ltd.), polyethylene glycol 6000 (produced by Wako Pure Chemical Industry Co., Ltd.) and NaCl and solubilized with 0.1 M NaOH and then subjected to SDS-PAGE. An about 135 kD band was cut out and pulverized to small pieces, which then were mixed with an adjuvant. The mixture was intradermally injected to a rabbit to immunize it.
[0061]To decrease the background upon screening a polypeptide gene clone that constitutes the parasporal body by using an antibody, absorption operation of the obtained antiserum with Escherichia coli fragments was repeated three times to remove the antibody that adsorbs on Escherichia coli itself and collect only the antibody that adsorbs on the polypeptide constituting the target parasporal body.
Example 2
[0062]A chromosomal DNA was collected from the bacterial cells of Bacillus popilliae semadara by an ordinary method and digested with restriction enzyme EcoRI. This was ligated to phosphatase-treated pBluescript SK(+) and Escherichia coli DH5α was transformed with the ligation product. The transformants were spread over the LB agar plate so that 2,000 to 3,000 colonies could appear thereon.
[0063]A nitrocellulose membrane was closely contacted on the agar plate to transfer colonies on the membrane and the membrane was placed on a fresh agar plate so that the colonies-attached side of the membrane directs upward. The plate was incubated at 37° C. for 2 hours. For bacteriolysis of the bacterial cells on the membrane, the following operation was performed. That is, the membrane was transferred on a filter paper impregnated with 10% SDS and left to stand for 5 minutes. Then, the membrane was transferred on a filter paper impregnated with 0.2 M NaOH, 0.15 M NaCl and 1% β-mercaptoethanol and left to stand for 5 minutes and finally it was transferred on a filter paper impregnated with 0.15 M NaCl, 0.5 M Tris-HCl, and pH 7.5, and left to stand for 10 minutes. The membrane was transferred to a buffer solution (0.1 M Tris-HCl, pH 7.8, 0.15 M NaCl, 5 mM MgCl2, 40 μg/ml lysozyme, and 5% skimmed milk) and shaken for 2 hours to remove the bacterial cells.
[0064]After washing the membrane with TBST (0.02 M Tris-HCl, pH 7.2, 0.5 M NaCl, and 0.05% Tween 20) for 5 minutes, the membrane was shaken for 1.5 hours in an anti-parasporal body antiserum solution diluted 8,000 folds with an antibody diluting liquid (TBST containing 5% skimmed milk). After repeating 10 minutes' washing with TBST three times, the membrane was transferred into a solution of 2.000-fold diluted Anti-Rabbit IgG (H&L), HRP-Linked (produced by New England Biolabs, Inc.) and incubated until color development was observed in an HRP coloring substrate (4CN, produced by NEN Life Science Products Co.).
[0065]Colonies near a site where strong coloring was observed, that is, to which the anti-parasporal body antiserum was adsorbed, were collected one by one from the master plate and the collected colonies were subjected to secondary screening performed in the same manner as described above to obtain a positive clone.
[0066]Plasmid was collected from the positive clone and the nucleotide sequence of the inserted portion was determined by an ordinary method. As a result, it revealed that the plasmid contained two polypeptide genes, which constitute the parasporal body of Bacillus popilliae, (SEQ ID NOS: 17 and 19, respective translation products being of SEQ ID NOS: 18 and 20) in a single cloned fragment. The amino acid sequence of SEQ ID NO: 18 and that of SEQ ID NO: 20 have about 73% of homology to each other. In this case, gene information processing software GENETYX-WIN (Software Development Co., Ltd.) is used for retrieval of the homology.
Example 3
[0067]PCR was performed by using the plasmid of the above-mentioned positive clone as a template and polynucleotides having nucleotide sequences corresponding to parts of a polynucleotide of SEQ ID NO: 19 and a complementary strand thereof (SEQ ID NOS: 21 and 22, respectively) as primers. After electrophoresing the reaction solution, about 4 kb of amplified fragments were recovered and purified and digested with restriction enzymes EcoRI and PstI. The digested products were ligated to a vector pTrc99A (produced by Amersham Pharmacia Biotech Co., Ltd.) that had been digested with EcoRI and PstI in advance, and Escherichia coli BL21(DE3) was transformed with the reaction solution. The transformants obtained were shake-cultured overnight in a 2×YT liquid medium containing 50 mg/l of ampicillin and then IPTG (isopropyl β-D-thiogalactopyranoside) was added so as to bring the transformants to a final concentration of 1 mM, followed by further culture for 2 hours. Observation with a microscope indicated formation of an inclusion body in the cells. Therefore, the clone was named as an expression clone (1).
Example 4
[0068]In a 2-liter baffled Erlenmeyer flask was charged 750 ml of a 2×YT liquid medium containing 50 mg/l of ampicillin and the expression clone (1) obtained in Example 3 was inoculated therein. The medium was shake-cultured at 37° C. 16 hours afterwards, IPTG is added to the medium so as to bring it to a final concentration of 1 mM, and the medium was further cultured for 2 hours. The cultured medium was centrifuged to recover from the culture solution bacterial cells, which were washed with a 10 mM potassium phosphate buffer solution, pH 7.0. Again the suspension was centrifuged to recover the bacterial cells, which were suspended in the above-mentioned buffer solution to finally make it into 120 ml. 10 g per cup of leaf mold was measured out and charged in 40 plastic-made food cups of 6 cm in diameter. In 20 cups out of them was dispersed 1 ml per cup of the suspension of the expression clone (1) and 1 ml per cup of water as blank was dispersed in the remaining 20 cups, followed by well mixing. In each of the food cups, one first instar larva of Anomala cuprea was released and fed under a condition of 25° C. Mortality of the larvae and mean body weight of the surviving larvae were measured with a lapse of time to verify the insecticidal activity and larvae growth inhibiting activity of the expression clone (1).
[0069]The results are shown in Table 1 and FIG. 2. A lot where the expression clone (1) was dispersed showed an increase in mortality, and on 5th week, high mortality of 95% was obtained. Further, the mean body weight of surviving larvae in the lot where the expression clone (1) was dispersed did not substantially increase but remained at a low level as compared with the blank. These results confirmed that the expression clone (1), that is, the polypeptide of the present invention has a larvae growth inhibiting or insecticidal activity.
TABLE-US-00001 TABLE 1 Cumulative Mortality (%) test lot 1st week 2nd week 3rd week 4th week 5th week Expression clone 5.0 45.0 65.0 80.0 95.0 Blank 5.0 5.0 5.0 5.0 5.0
Example 5
[0070]In a 2-liter baffled Erlenmeyer flask was charged 750 ml of a 2×YT liquid medium containing 50 mg/l of ampicillin and the expression clone (1) obtained in Example 3 was inoculated therein. The medium was shake-cultured at 37° C. After 16 hours, IPTG was added so as to bring the medium to a final concentration of 1 mM and the medium was further cultured for 2 hours. The culture medium was centrifuged to recover from the culture solution bacterial cells, which were washed with a 10 mM potassium phosphate buffer solution, pH 7.0. Again the suspension was centrifuged to recover the bacterial cells, which were suspended in the above-mentioned buffer solution to finally make it into 75 ml, thus making it as an expression clone suspension.
[0071]On the other hand, the sporangia of Bacillus popilliae semadara were collected from larvae of Anomala cuprea infected with milky disease and washed with water. They were suspended again in water to a density of 8×107 sporangia/ml to make it as a sporangia suspension.
[0072]80 plastic-made food cups of 6 cm in diameter, each of which was charged with 25 g of leaf mold that was measured out, were prepared. In 20 cups out of them was dispersed 1 ml per cup of the suspension of the expression clone (1), in other 20 cups was dispersed 0.5 ml per cup of the sporangia suspension, in still other 20 cups was dispersed 1 ml of the expression clone suspension together with 0.5 ml of the sporangia suspension, and in the remaining 20 cups was dispersed 1 ml per cup of water, followed by well mixing. In each of the food cups, a second instar larva of Anomala cuprea was released one by one and fed under a condition of 25° C. for 21 days. Mortality of the larvae and mean body weight of the surviving larvae were measured with a lapse of time to verify synergistic effects in the insecticidal activity and the larvae growth inhibiting activity of the expression clone (1) used in combination with the sporangia.
[0073]The results are shown in FIGS. 3 and 4. Dispersion of the suspension of the expression clone (1) or the sporangia alone exhibited a larvae growth inhibiting effect but did not exhibit a sufficient insecticidal effect. In contrast, when the expression clone (1) and the sporangia were dispersed in combination, an abrupt increase in mortality was observed, so that synergistic effect was confirmed.
Example 6
[0074]Polynucleotides having the nucleotide sequences of SEQ ID NO: 11 and SEQ ID NO: 13 corresponding to parts of complementary strands of SEQ ID NO: 17 and SEQ ID NO: 19, respectively, were prepared by chemical synthesis. PCR performed by using the polynucleotides as primers and a chromosomal DNA of Bacillus popilliae semadara as a template indicated amplification of about 4.2 kb fragments. TA cloning of the fragments gave rise to a number of clones, some of which were subjected to determination of the nucleotide sequence of the insert portion by sequencing. As a result, polynucleotides having the nucleotide sequences of SEQ ID NO: 17 and SEQ ID NO: 19 were obtained. Therefore, it revealed that the polynucleotide of the present invention can also be obtained by PCR using the polynucleotides having the nucleotide sequences of SEQ ID NO: 11 and SEQ ID NO: 13, respectively.
Example 7
[0075]A chromosomal DNA was collected from the bacterial cells of Bacillus popilliae var. popilliae Mame by an ordinary method. On the other hand, based on the nucleotide sequence of the gene encoding the polypeptide constituting the parasporal body of Bacillus popilliae semadara as a reference, polynucleotides having the nucleotide sequences of SEQ ID NO: 11 and SEQ ID NO: 14, respectively, were chemically synthesized. PCR performed using the polynucleotides as primers and the chromosomal DNA as a template indicated amplification of about 4.3 kb fragments. TA cloning of the fragments gave rise to a number of clones, one of which was subjected to determination of the nucleotide sequence of the insert by sequencing. As a result, a polynucleotide having the nucleotide sequence of SEQ ID NO: 3 was obtained. This was further translated to obtain the amino acid sequence of SEQ ID NO: 4.
Example 8
[0076]Polynucleotides having nucleotide sequences of SEQ ID NO: 12 and SEQ ID NO: 13, respectively, were prepared by chemical synthesis. PCR performed by using the polynucleotides as primers and the chromosomal DNA prepared as described above as a template indicated amplification of about 4.2 kb fragments. TA cloning of the fragments gave rise to a large number of clones, one of which was subjected to determination of the nucleotide sequence of the insert portion by sequencing. As a result, a polynucleotide having the nucleotide sequence of SEQ ID NO: 5 was obtained. This was further translated to obtain the amino acid sequence of SEQ ID NO: 6.
Example 9
[0077]Polynucleotides having the nucleotide sequence of SEQ ID NO: 15 corresponding to a part of a complementary strand of SEQ ID NO: 3 and the nucleotide sequence of SEQ ID NO: 11, respectively, were prepared by chemical synthesis. PCR performed by using the polynucleotides as primers and the chromosomal DNA prepared as described above as a template indicated amplification of about 1.2 kb fragments. TA cloning of the fragments gave rise to a large number of clones, one of which was subjected to determination of the nucleotide sequence of the insert portion by sequencing. As a result, a polynucleotide having the nucleotide sequence of SEQ ID NO: 7 was obtained. This was further translated to obtain the amino acid sequence of SEQ ID NO: 8.
Example 10
[0078]The chromosomal DNA obtained as described above was degested with restriction enzyme XhoI and the fragments were ligated with SalI Cassette contained in a LA PCR in vitro Cloning Kit produced by Takara Shuzo Co., Ltd. On the other hand, a polynucleotide having the nucleotide sequence (SEQ ID NO: 16) corresponding to a part of a sequence of SEQ ID NO: 3 was prepared by chemical synthesis. Then, PCR was performed by using the polynucleotide and a Cassette primer C1 attached to the Kit as primers and the ligated chromosomal DNA fragment as a template. As a result, amplification of about 3 kb fragments was observed. TA cloning of the fragments gave rise to a large number of clones, one of which was subjected to determination of the nucleotide sequence of the insert portion by sequencing. As a result, a clone having a polynucleotide having the nucleotide sequence of SEQ ID NO: 9 coupled in tandem to downstream of the latter half of the nucleotide sequence of SEQ ID NO: 3 was obtained. The nucleotide sequence of SEQ ID NO: 9 was further translated to obtain the amino acid sequence of SEQ ID NO: 10.
Example 11
[0079]The amino acid sequences of the polypeptides of the parasporal bodies existing in Bacillus popilliae obtained in Examples 2, 7 and 8 and the nucleotide sequences of the polynucleotides encoding them were comparatively studied by using gene information processing software GENETYX-WIN (Software Development Co., Ltd.). As a result, sequences that have very high homology of 98% or more in the amino acid sequence and are considered to be specific to the parasporal body of Bacillus popilliae, i.e., the amino acid sequence of SEQ ID NO: 2 and the corresponding nucleotide sequence of SEQ ID NO: 1 were found. The portions indicated by an arrow in FIGS. 5 to 7 correspond to the amino acid sequence of SEQ ID NO: 2.
[0080]Note that these sequences have homology of 9% or less in the amino acid sequence to the sequence isolated from Bacillus popilliae subsp. Melolonthae H1 described in WO97/14798, and hence is completely different therefrom.
Example 12
[0081]PCR was performed by using a chromosomal DNA of Bacillus popilliae var. popilliae Mame as a template and polynucleotides of SEQ ID NO: 12 and SEQ ID NO: 14 as primers to amplify about 4 kb fragments, which were purified and recovered by electrophoresis and subsequent cutting out. The fragments were ligated to a pT7Blue vector (Novagen, Inc.) and Escherichia coli DH5α was transformed with the ligation product. Among the obtained clones, a clone harboring the pT7Blue with the 4 kb insert downstream of lac promoter in a direction of sense to form a fusion polypeptide with an N-terminal portion of a lacZ polypeptide was selected. This was named as an expression clone (2) and used in the following tests of production of a polypeptide and effects thereof.
Example 13
[0082]Ten 300 ml-capacity Erlenmeyer flasks containing 200 ml of a 2×YT medium were prepared. In 5 flasks out of them was inoculated the expression clone (2) obtained in Example 12 and shake-cultured at 37° C. overnight. As a comparative control clone, Escherichia coli DH5α having a pT7Blue vector having inserted therein no 4 kb fragment was inoculated in the remaining 5 flasks and cultured in the same manner as the above. After completion of the culture, bacterial cells were observed with a microscope. As a result, formation of inclusion body in the bacterial cells was observed only in the case of the expression clone (2). The bacterial cells were once recovered from each flask by centrifugation and after washing with water, the bacterial cells were centrifuged again to recover them and then suspended in water to finally make it into 23 ml.
[0083]20 g per cup of leaf mold was measured out and charged in 15 plastic-made food cups of 6 cm in diameter. In 5 cups out of them was added a suspension of the expression clone and in other 5 cups out of them was added a suspension of the comparative control clone, followed by well mixing. In the remaining 5 cups was dedicated to blank tests of no addition. In each of the food cups, one first instar larva of Anomala cuprea was released and fed under a condition of 25° C. The body weights of larvae were measured with a lapse of time and compared to verify the larvae growth inhibiting activity of the expression clone (2). In contrast to the blank test and the control lot, a lot where the expression clone (2) was added apparently suppressed increase in mean body weight (FIG. 8), which confirmed that the expression clone (2), that is, the polypeptide of the present invention has a larvae growth inhibiting effect.
Sequence CWU
1
SEQUENCE LISTING
<160> NUMBER OF SEQ ID NOS: 22
<210> SEQ ID NO 1
<211> LENGTH: 402
<212> TYPE: DNA
<213> ORGANISM: Bacillus popilliae
<220> FEATURE:
<221> NAME/KEY: CDS
<222> LOCATION: (1)..(402)
<400> SEQUENCE: 1
aga aaa cac cgc aaa tgt cat caa gcg cat caa ttt gag ttc cat att 48
Arg Lys His Arg Lys Cys His Gln Ala His Gln Phe Glu Phe His Ile
1 5 10 15
gat acc ggg aca atc gat ctg gtc gaa gat ttg ggc att tgg gtg atc 96
Asp Thr Gly Thr Ile Asp Leu Val Glu Asp Leu Gly Ile Trp Val Ile
20 25 30
ttc aaa atc tgt gcc aca gat ggt tac gca agc tta gat gat ttg gaa 144
Phe Lys Ile Cys Ala Thr Asp Gly Tyr Ala Ser Leu Asp Asp Leu Glu
35 40 45
gtg att gaa gaa gga gcg ctg ggt gtc gaa gca tta gaa ctt gtc aag 192
Val Ile Glu Glu Gly Ala Leu Gly Val Glu Ala Leu Glu Leu Val Lys
50 55 60
aaa aga gaa aag aaa tgg aga cat cag aag gag cag cac tgt tcg caa 240
Lys Arg Glu Lys Lys Trp Arg His Gln Lys Glu Gln His Cys Ser Gln
65 70 75 80
acg aaa cac aaa tat gat gcg gcc aaa cac gcg gtg atg gcg tta ttc 288
Thr Lys His Lys Tyr Asp Ala Ala Lys His Ala Val Met Ala Leu Phe
85 90 95
aca aac acg cgc tat gaa aaa ttg aag ttc gaa aca acc atc tcc aat 336
Thr Asn Thr Arg Tyr Glu Lys Leu Lys Phe Glu Thr Thr Ile Ser Asn
100 105 110
att ttg tat gct gat cat ctc gtg cag tcg att cct tat gta tat aat 384
Ile Leu Tyr Ala Asp His Leu Val Gln Ser Ile Pro Tyr Val Tyr Asn
115 120 125
aaa tat gta ccg gaa gtt 402
Lys Tyr Val Pro Glu Val
130
<210> SEQ ID NO 2
<211> LENGTH: 134
<212> TYPE: PRT
<213> ORGANISM: Bacillus popilliae
<400> SEQUENCE: 2
Arg Lys His Arg Lys Cys His Gln Ala His Gln Phe Glu Phe His Ile
1 5 10 15
Asp Thr Gly Thr Ile Asp Leu Val Glu Asp Leu Gly Ile Trp Val Ile
20 25 30
Phe Lys Ile Cys Ala Thr Asp Gly Tyr Ala Ser Leu Asp Asp Leu Glu
35 40 45
Val Ile Glu Glu Gly Ala Leu Gly Val Glu Ala Leu Glu Leu Val Lys
50 55 60
Lys Arg Glu Lys Lys Trp Arg His Gln Lys Glu Gln His Cys Ser Gln
65 70 75 80
Thr Lys His Lys Tyr Asp Ala Ala Lys His Ala Val Met Ala Leu Phe
85 90 95
Thr Asn Thr Arg Tyr Glu Lys Leu Lys Phe Glu Thr Thr Ile Ser Asn
100 105 110
Ile Leu Tyr Ala Asp His Leu Val Gln Ser Ile Pro Tyr Val Tyr Asn
115 120 125
Lys Tyr Val Pro Glu Val
130
<210> SEQ ID NO 3
<211> LENGTH: 4359
<212> TYPE: DNA
<213> ORGANISM: Bacillus popilliae
<220> FEATURE:
<221> NAME/KEY: CDS
<222> LOCATION: (282)..(4229)
<400> SEQUENCE: 3
gctgcattcc ctttatcatc aatattgaag tcaaaccgaa gggaaatcgt gaaacatgtt 60
ttactgaccc ccatgccaaa gaaatctcac tttgttgtga ggggagtgta tgtgtagacc 120
atgtactcaa atgcagtgtc ggaagtttgc cagatgttca tatcaattgt aaatatgtga 180
cggtatgtga tttacagatg aggcccgttc atgagggagc ttgtcaattt gtgaagatta 240
gcggagaatt tcaattttat tcactttaac attagctaca a atg ttg tgg cta aat 296
Met Leu Trp Leu Asn
1 5
act cat tgt tca tta ata gga ata act ggg agg caa act atg aat caa 344
Thr His Cys Ser Leu Ile Gly Ile Thr Gly Arg Gln Thr Met Asn Gln
10 15 20
tat cat aac caa aac gat aac aaa agt tac aac caa agt gga aat gaa 392
Tyr His Asn Gln Asn Asp Asn Lys Ser Tyr Asn Gln Ser Gly Asn Glu
25 30 35
atg caa atc att caa cct tca agt aac gct tta ctt tac agt ccc aat 440
Met Gln Ile Ile Gln Pro Ser Ser Asn Ala Leu Leu Tyr Ser Pro Asn
40 45 50
aag tat ccg tat gct acg gat ccc aat gtc ata gca gag ggt gga agt 488
Lys Tyr Pro Tyr Ala Thr Asp Pro Asn Val Ile Ala Glu Gly Gly Ser
55 60 65
tat aaa aat tgg ttg gat atg tgt aca ggg aca ggc gac aca cga agc 536
Tyr Lys Asn Trp Leu Asp Met Cys Thr Gly Thr Gly Asp Thr Arg Ser
70 75 80 85
ccc gaa act gct gct att tca aaa ggt gct gtt tct gct gca att act 584
Pro Glu Thr Ala Ala Ile Ser Lys Gly Ala Val Ser Ala Ala Ile Thr
90 95 100
ata agc acc ggg ctt ctt ggc tta cta ggt gtt ccg ttt gca tca caa 632
Ile Ser Thr Gly Leu Leu Gly Leu Leu Gly Val Pro Phe Ala Ser Gln
105 110 115
atc ggg gca ttt tat acc ttc cta ttg aat acc tta tgg cct gca agc 680
Ile Gly Ala Phe Tyr Thr Phe Leu Leu Asn Thr Leu Trp Pro Ala Ser
120 125 130
aat act caa tgg gag cag ttt ata gca cat gtg gaa gaa ctc ata aat 728
Asn Thr Gln Trp Glu Gln Phe Ile Ala His Val Glu Glu Leu Ile Asn
135 140 145
gca aaa cta aca gat cat gta aga aat tcg gca ctt aca aaa tta aat 776
Ala Lys Leu Thr Asp His Val Arg Asn Ser Ala Leu Thr Lys Leu Asn
150 155 160 165
ggt tta cgt aat aac ata gag ata tat aac gaa gct tta ata gtt tgg 824
Gly Leu Arg Asn Asn Ile Glu Ile Tyr Asn Glu Ala Leu Ile Val Trp
170 175 180
aaa caa gat cct aac aat agc aaa cta aaa gat gat gta aga agt aaa 872
Lys Gln Asp Pro Asn Asn Ser Lys Leu Lys Asp Asp Val Arg Ser Lys
185 190 195
ttc gta ggt cta aat agc caa ttc gaa gaa tat att cca caa ttt aaa 920
Phe Val Gly Leu Asn Ser Gln Phe Glu Glu Tyr Ile Pro Gln Phe Lys
200 205 210
gaa gaa ggt ttt gaa gtt caa tta tta act ata tat gca caa tct gca 968
Glu Glu Gly Phe Glu Val Gln Leu Leu Thr Ile Tyr Ala Gln Ser Ala
215 220 225
aat ctt cat cta tta tta tta aga gat tcc tct ttg tat ggt gca tct 1016
Asn Leu His Leu Leu Leu Leu Arg Asp Ser Ser Leu Tyr Gly Ala Ser
230 235 240 245
tgg gga ttt gct caa gct act att gac aat aat tac aat cgc caa ata 1064
Trp Gly Phe Ala Gln Ala Thr Ile Asp Asn Asn Tyr Asn Arg Gln Ile
250 255 260
agg aaa acc gca gag tat gca aat cat tgt acc act tgg tat cag acg 1112
Arg Lys Thr Ala Glu Tyr Ala Asn His Cys Thr Thr Trp Tyr Gln Thr
265 270 275
ggt tta caa aga ttg caa ggc act act gct agc agt tgg ctc tct tat 1160
Gly Leu Gln Arg Leu Gln Gly Thr Thr Ala Ser Ser Trp Leu Ser Tyr
280 285 290
cat aga ttt aga aga gaa atg aca cta aca gta ttg gat att tgc gca 1208
His Arg Phe Arg Arg Glu Met Thr Leu Thr Val Leu Asp Ile Cys Ala
295 300 305
tta ttt tca aat tat gat gcc cgt agt tac cca ctg gag gta agg gga 1256
Leu Phe Ser Asn Tyr Asp Ala Arg Ser Tyr Pro Leu Glu Val Arg Gly
310 315 320 325
gag ctt acg aga gaa att tat acg gat cca gta gca ccc ggt act aac 1304
Glu Leu Thr Arg Glu Ile Tyr Thr Asp Pro Val Ala Pro Gly Thr Asn
330 335 340
tgg ata gat cga gca cca tca ttc gca gaa ata gaa aat cta gta att 1352
Trp Ile Asp Arg Ala Pro Ser Phe Ala Glu Ile Glu Asn Leu Val Ile
345 350 355
agg gca cca aga act gtt act tgg ata tcc ggt gat tta ata gta tat 1400
Arg Ala Pro Arg Thr Val Thr Trp Ile Ser Gly Asp Leu Ile Val Tyr
360 365 370
aca ggt aga ttg tac ggc tat act ggt aat aac gat tat tgg gca gca 1448
Thr Gly Arg Leu Tyr Gly Tyr Thr Gly Asn Asn Asp Tyr Trp Ala Ala
375 380 385
cac agg cta gat ttt ctt gaa acc aat ggt tat cgg ttt gag ggt cct 1496
His Arg Leu Asp Phe Leu Glu Thr Asn Gly Tyr Arg Phe Glu Gly Pro
390 395 400 405
acc tat gga tcg acg att aat ata agt cgt aca gat tct att ccc atg 1544
Thr Tyr Gly Ser Thr Ile Asn Ile Ser Arg Thr Asp Ser Ile Pro Met
410 415 420
aat tct att gat gtt tat tcc act act gta gtg act gtt ggc tct gct 1592
Asn Ser Ile Asp Val Tyr Ser Thr Thr Val Val Thr Val Gly Ser Ala
425 430 435
tgg cca act ggc ggt ttt gtg ttg gga gtc gct tcg gct aga ttt ttt 1640
Trp Pro Thr Gly Gly Phe Val Leu Gly Val Ala Ser Ala Arg Phe Phe
440 445 450
tcg aaa agt cct agc acc ggt tta tta ggt gag cgg gtg tat cag aat 1688
Ser Lys Ser Pro Ser Thr Gly Leu Leu Gly Glu Arg Val Tyr Gln Asn
455 460 465
cca gta tat ttt tcg agt tcc act tta act ttt aac tta cct gga gta 1736
Pro Val Tyr Phe Ser Ser Ser Thr Leu Thr Phe Asn Leu Pro Gly Val
470 475 480 485
gac caa gat acg cca act gct gcc gac tat agt cat aaa cta tcg tgt 1784
Asp Gln Asp Thr Pro Thr Ala Ala Asp Tyr Ser His Lys Leu Ser Cys
490 495 500
atc aca gca ttt cga act gga ttg aat gga act gtt ccg gtt ttt gga 1832
Ile Thr Ala Phe Arg Thr Gly Leu Asn Gly Thr Val Pro Val Phe Gly
505 510 515
cgg tat tct gca act gtt agt cgt gac aat aga att gag cca gac aaa 1880
Arg Tyr Ser Ala Thr Val Ser Arg Asp Asn Arg Ile Glu Pro Asp Lys
520 525 530
ata acg caa atc ccg gct gtt aag tca aac tcc ctc gac aat tgt cca 1928
Ile Thr Gln Ile Pro Ala Val Lys Ser Asn Ser Leu Asp Asn Cys Pro
535 540 545
gta gtt aga ggg act gga ttt aca gga gga gac tgg ttg aag aca agt 1976
Val Val Arg Gly Thr Gly Phe Thr Gly Gly Asp Trp Leu Lys Thr Ser
550 555 560 565
tat ctt agt gtt ttt gtc cta act atc act tca tcg aga gcg ggc caa 2024
Tyr Leu Ser Val Phe Val Leu Thr Ile Thr Ser Ser Arg Ala Gly Gln
570 575 580
tct tac cgc atc cgc gtc cgt tat gct gct gca gta gat tta att atg 2072
Ser Tyr Arg Ile Arg Val Arg Tyr Ala Ala Ala Val Asp Leu Ile Met
585 590 595
agt ata tat tct aat gac cct ttt att tcc aaa gga att agt ctt acc 2120
Ser Ile Tyr Ser Asn Asp Pro Phe Ile Ser Lys Gly Ile Ser Leu Thr
600 605 610
aaa tca atg cca cca ctg acc gaa act gta cct tac gaa gct ttt aaa 2168
Lys Ser Met Pro Pro Leu Thr Glu Thr Val Pro Tyr Glu Ala Phe Lys
615 620 625
ttt gca gat ttt ggt gtc act ttt aca aca gct act gct aac aaa aga 2216
Phe Ala Asp Phe Gly Val Thr Phe Thr Thr Ala Thr Ala Asn Lys Arg
630 635 640 645
tat act ttt caa ttc cat acg ggt gga gca gct ata att gac aga att 2264
Tyr Thr Phe Gln Phe His Thr Gly Gly Ala Ala Ile Ile Asp Arg Ile
650 655 660
gaa ttt gtt cca att gag ggt agt ttg ttc gag tac gaa acc aaa caa 2312
Glu Phe Val Pro Ile Glu Gly Ser Leu Phe Glu Tyr Glu Thr Lys Gln
665 670 675
cag cta gaa aaa gca agg aaa gcg gtg aac cat ttg ttt aca gat gga 2360
Gln Leu Glu Lys Ala Arg Lys Ala Val Asn His Leu Phe Thr Asp Gly
680 685 690
tcg aaa aag gcg cta aaa gaa ggc acg acc gat tat gag atc gat caa 2408
Ser Lys Lys Ala Leu Lys Glu Gly Thr Thr Asp Tyr Glu Ile Asp Gln
695 700 705
gcc gcc aac gtg gtg gat tgt ata tcg gat gag tgt gga cat gag aaa 2456
Ala Ala Asn Val Val Asp Cys Ile Ser Asp Glu Cys Gly His Glu Lys
710 715 720 725
atg atc ctg tta gat gaa gta aaa tat gca aaa caa ctc agc caa gcc 2504
Met Ile Leu Leu Asp Glu Val Lys Tyr Ala Lys Gln Leu Ser Gln Ala
730 735 740
cgc aat tta ctg ctc aat ggg aat ttc gat gat cta tat cca gct ctg 2552
Arg Asn Leu Leu Leu Asn Gly Asn Phe Asp Asp Leu Tyr Pro Ala Leu
745 750 755
gag agg gag aat cca tgg aaa aca agt ccg cat gtt acg atc cgt caa 2600
Glu Arg Glu Asn Pro Trp Lys Thr Ser Pro His Val Thr Ile Arg Gln
760 765 770
gat aac ccg att ttt aaa ggc cat tat ctc agt atg gcg ggt gcg aac 2648
Asp Asn Pro Ile Phe Lys Gly His Tyr Leu Ser Met Ala Gly Ala Asn
775 780 785
gat att gag gcc acc aat gat acc ttc ccc acg tat gtc tat caa aaa 2696
Asp Ile Glu Ala Thr Asn Asp Thr Phe Pro Thr Tyr Val Tyr Gln Lys
790 795 800 805
ata gac gaa gcc aaa tta aag cca tat aca cgg tat aaa gtg cgc ggg 2744
Ile Asp Glu Ala Lys Leu Lys Pro Tyr Thr Arg Tyr Lys Val Arg Gly
810 815 820
ttt gtt ggt agc agc aaa gct cta gag ctg ttg gtt aca cgc tat aat 2792
Phe Val Gly Ser Ser Lys Ala Leu Glu Leu Leu Val Thr Arg Tyr Asn
825 830 835
gaa gaa gtc gat gcg att tta gat gta ccg gat aat atc ccg cat gcg 2840
Glu Glu Val Asp Ala Ile Leu Asp Val Pro Asp Asn Ile Pro His Ala
840 845 850
ccg act cct gtc tgc ggt gaa ttt gat cga tgc aag ccc tat tcg tat 2888
Pro Thr Pro Val Cys Gly Glu Phe Asp Arg Cys Lys Pro Tyr Ser Tyr
855 860 865
cca cct tta ctt cca gaa tgt aac cct gag ttt ata aat cag atg caa 2936
Pro Pro Leu Leu Pro Glu Cys Asn Pro Glu Phe Ile Asn Gln Met Gln
870 875 880 885
cca tcc tct tgc cac cac aat cag atg gtc gat tac aat aac aga aaa 2984
Pro Ser Ser Cys His His Asn Gln Met Val Asp Tyr Asn Asn Arg Lys
890 895 900
cac cgc aaa tgt cat caa gcg cat caa ttt gag ttc cat att gat acc 3032
His Arg Lys Cys His Gln Ala His Gln Phe Glu Phe His Ile Asp Thr
905 910 915
ggg aca atc gat ctg gtc gaa gat ttg ggc att tgg gtg atc ttc aaa 3080
Gly Thr Ile Asp Leu Val Glu Asp Leu Gly Ile Trp Val Ile Phe Lys
920 925 930
atc tgt gcc aca gat ggt tac gca agc tta gat gat ttg gaa gtg att 3128
Ile Cys Ala Thr Asp Gly Tyr Ala Ser Leu Asp Asp Leu Glu Val Ile
935 940 945
gaa gaa gga gcg ctg ggt gtc gaa gca tta gaa ctt gtc aag aaa aga 3176
Glu Glu Gly Ala Leu Gly Val Glu Ala Leu Glu Leu Val Lys Lys Arg
950 955 960 965
gaa aag aaa tgg aga cat cag aag gag cag cac tgt tcg caa acg aaa 3224
Glu Lys Lys Trp Arg His Gln Lys Glu Gln His Cys Ser Gln Thr Lys
970 975 980
cac aaa tat gat gcg gcc aaa cac gcg gtg atg gcg tta ttc aca aac 3272
His Lys Tyr Asp Ala Ala Lys His Ala Val Met Ala Leu Phe Thr Asn
985 990 995
acg cgc tat gaa aaa ttg aag ttc gaa aca acc atc tcc aat att ttg 3320
Thr Arg Tyr Glu Lys Leu Lys Phe Glu Thr Thr Ile Ser Asn Ile Leu
1000 1005 1010
tat gct gat cat ctc gtg cag tcg att cct tat gta tat aat aaa tat 3368
Tyr Ala Asp His Leu Val Gln Ser Ile Pro Tyr Val Tyr Asn Lys Tyr
1015 1020 1025
gta ccg gaa gtt cca ggt atg aat tac gaa ctc tat aca gag cta aac 3416
Val Pro Glu Val Pro Gly Met Asn Tyr Glu Leu Tyr Thr Glu Leu Asn
1030 1035 1040 1045
act ctc gtt cag aat gcg ttc tat cta tat gac cag cgg aat ctg att 3464
Thr Leu Val Gln Asn Ala Phe Tyr Leu Tyr Asp Gln Arg Asn Leu Ile
1050 1055 1060
aaa aat ggg cgc ttt agc aat ggg ctt atg tat tgg cag gct acc ccg 3512
Lys Asn Gly Arg Phe Ser Asn Gly Leu Met Tyr Trp Gln Ala Thr Pro
1065 1070 1075
cat gca cga gtg gaa caa gaa tat gag aaa tct gta ctc gtg ctg cca 3560
His Ala Arg Val Glu Gln Glu Tyr Glu Lys Ser Val Leu Val Leu Pro
1080 1085 1090
aat tgg gat gcc aat gtg tcg caa gat ctt tgt atc gaa cac aat cgc 3608
Asn Trp Asp Ala Asn Val Ser Gln Asp Leu Cys Ile Glu His Asn Arg
1095 1100 1105
ggt tat gta ttg cgt gtc acg gcg aga aaa gaa gat ccg gga gct ggc 3656
Gly Tyr Val Leu Arg Val Thr Ala Arg Lys Glu Asp Pro Gly Ala Gly
1110 1115 1120 1125
aat gtt acc ttc agt gac tgt gca aac cat gtc gac aag ctg agc ttt 3704
Asn Val Thr Phe Ser Asp Cys Ala Asn His Val Asp Lys Leu Ser Phe
1130 1135 1140
act tct tgc gat ata gct aca aac gca gtg cca ggt gcc caa gcg aat 3752
Thr Ser Cys Asp Ile Ala Thr Asn Ala Val Pro Gly Ala Gln Ala Asn
1145 1150 1155
gat cca gcc gcc gga gta gcc tat gga caa cag ggc tgt caa ata gat 3800
Asp Pro Ala Ala Gly Val Ala Tyr Gly Gln Gln Gly Cys Gln Ile Asp
1160 1165 1170
aga gtg ccg tac gga caa tct gga tat aga aca gac gga aca aat gga 3848
Arg Val Pro Tyr Gly Gln Ser Gly Tyr Arg Thr Asp Gly Thr Asn Gly
1175 1180 1185
atg ccg tac gga cag tct ggt aat cga gca gac ggg gtg cca tac aga 3896
Met Pro Tyr Gly Gln Ser Gly Asn Arg Ala Asp Gly Val Pro Tyr Arg
1190 1195 1200 1205
caa tcc gga tat gga aca gat gga gta gcg cac gac caa cct gga tat 3944
Gln Ser Gly Tyr Gly Thr Asp Gly Val Ala His Asp Gln Pro Gly Tyr
1210 1215 1220
cga gca gac gga gta gcg tac gaa caa tct gga tat cga gca gac gga 3992
Arg Ala Asp Gly Val Ala Tyr Glu Gln Ser Gly Tyr Arg Ala Asp Gly
1225 1230 1235
gta acg tat gac caa tct gcc aat caa acc cgc aaa tat cat ggt tgc 4040
Val Thr Tyr Asp Gln Ser Ala Asn Gln Thr Arg Lys Tyr His Gly Cys
1240 1245 1250
cat aca gtc gga ctg cca cat cca gag cat ggt tgt tgt tat cca gac 4088
His Thr Val Gly Leu Pro His Pro Glu His Gly Cys Cys Tyr Pro Asp
1255 1260 1265
aga gta agc gat ggc caa cag ctt gca tat gta aca aaa tcg att gat 4136
Arg Val Ser Asp Gly Gln Gln Leu Ala Tyr Val Thr Lys Ser Ile Asp
1270 1275 1280 1285
ctg ttc ccg gat aca gat aaa gtc cgg atc gac att gga gaa acc gaa 4184
Leu Phe Pro Asp Thr Asp Lys Val Arg Ile Asp Ile Gly Glu Thr Glu
1290 1295 1300
ggg aac ttt aga gtg gaa agt gtg gaa ttg att tgt atg gaa aag 4229
Gly Asn Phe Arg Val Glu Ser Val Glu Leu Ile Cys Met Glu Lys
1305 1310 1315
taaatcatca caagtaaaag tatcgtttac taaaaattta ttttccaagc aacaggggag 4289
aagatgattt gggtgtaata ctcaaaccat cttttcttat aagccacttt atgatcattc 4349
gtgatcgaga 4359
<210> SEQ ID NO 4
<211> LENGTH: 1316
<212> TYPE: PRT
<213> ORGANISM: Bacillus popilliae
<400> SEQUENCE: 4
Met Leu Trp Leu Asn Thr His Cys Ser Leu Ile Gly Ile Thr Gly Arg
1 5 10 15
Gln Thr Met Asn Gln Tyr His Asn Gln Asn Asp Asn Lys Ser Tyr Asn
20 25 30
Gln Ser Gly Asn Glu Met Gln Ile Ile Gln Pro Ser Ser Asn Ala Leu
35 40 45
Leu Tyr Ser Pro Asn Lys Tyr Pro Tyr Ala Thr Asp Pro Asn Val Ile
50 55 60
Ala Glu Gly Gly Ser Tyr Lys Asn Trp Leu Asp Met Cys Thr Gly Thr
65 70 75 80
Gly Asp Thr Arg Ser Pro Glu Thr Ala Ala Ile Ser Lys Gly Ala Val
85 90 95
Ser Ala Ala Ile Thr Ile Ser Thr Gly Leu Leu Gly Leu Leu Gly Val
100 105 110
Pro Phe Ala Ser Gln Ile Gly Ala Phe Tyr Thr Phe Leu Leu Asn Thr
115 120 125
Leu Trp Pro Ala Ser Asn Thr Gln Trp Glu Gln Phe Ile Ala His Val
130 135 140
Glu Glu Leu Ile Asn Ala Lys Leu Thr Asp His Val Arg Asn Ser Ala
145 150 155 160
Leu Thr Lys Leu Asn Gly Leu Arg Asn Asn Ile Glu Ile Tyr Asn Glu
165 170 175
Ala Leu Ile Val Trp Lys Gln Asp Pro Asn Asn Ser Lys Leu Lys Asp
180 185 190
Asp Val Arg Ser Lys Phe Val Gly Leu Asn Ser Gln Phe Glu Glu Tyr
195 200 205
Ile Pro Gln Phe Lys Glu Glu Gly Phe Glu Val Gln Leu Leu Thr Ile
210 215 220
Tyr Ala Gln Ser Ala Asn Leu His Leu Leu Leu Leu Arg Asp Ser Ser
225 230 235 240
Leu Tyr Gly Ala Ser Trp Gly Phe Ala Gln Ala Thr Ile Asp Asn Asn
245 250 255
Tyr Asn Arg Gln Ile Arg Lys Thr Ala Glu Tyr Ala Asn His Cys Thr
260 265 270
Thr Trp Tyr Gln Thr Gly Leu Gln Arg Leu Gln Gly Thr Thr Ala Ser
275 280 285
Ser Trp Leu Ser Tyr His Arg Phe Arg Arg Glu Met Thr Leu Thr Val
290 295 300
Leu Asp Ile Cys Ala Leu Phe Ser Asn Tyr Asp Ala Arg Ser Tyr Pro
305 310 315 320
Leu Glu Val Arg Gly Glu Leu Thr Arg Glu Ile Tyr Thr Asp Pro Val
325 330 335
Ala Pro Gly Thr Asn Trp Ile Asp Arg Ala Pro Ser Phe Ala Glu Ile
340 345 350
Glu Asn Leu Val Ile Arg Ala Pro Arg Thr Val Thr Trp Ile Ser Gly
355 360 365
Asp Leu Ile Val Tyr Thr Gly Arg Leu Tyr Gly Tyr Thr Gly Asn Asn
370 375 380
Asp Tyr Trp Ala Ala His Arg Leu Asp Phe Leu Glu Thr Asn Gly Tyr
385 390 395 400
Arg Phe Glu Gly Pro Thr Tyr Gly Ser Thr Ile Asn Ile Ser Arg Thr
405 410 415
Asp Ser Ile Pro Met Asn Ser Ile Asp Val Tyr Ser Thr Thr Val Val
420 425 430
Thr Val Gly Ser Ala Trp Pro Thr Gly Gly Phe Val Leu Gly Val Ala
435 440 445
Ser Ala Arg Phe Phe Ser Lys Ser Pro Ser Thr Gly Leu Leu Gly Glu
450 455 460
Arg Val Tyr Gln Asn Pro Val Tyr Phe Ser Ser Ser Thr Leu Thr Phe
465 470 475 480
Asn Leu Pro Gly Val Asp Gln Asp Thr Pro Thr Ala Ala Asp Tyr Ser
485 490 495
His Lys Leu Ser Cys Ile Thr Ala Phe Arg Thr Gly Leu Asn Gly Thr
500 505 510
Val Pro Val Phe Gly Arg Tyr Ser Ala Thr Val Ser Arg Asp Asn Arg
515 520 525
Ile Glu Pro Asp Lys Ile Thr Gln Ile Pro Ala Val Lys Ser Asn Ser
530 535 540
Leu Asp Asn Cys Pro Val Val Arg Gly Thr Gly Phe Thr Gly Gly Asp
545 550 555 560
Trp Leu Lys Thr Ser Tyr Leu Ser Val Phe Val Leu Thr Ile Thr Ser
565 570 575
Ser Arg Ala Gly Gln Ser Tyr Arg Ile Arg Val Arg Tyr Ala Ala Ala
580 585 590
Val Asp Leu Ile Met Ser Ile Tyr Ser Asn Asp Pro Phe Ile Ser Lys
595 600 605
Gly Ile Ser Leu Thr Lys Ser Met Pro Pro Leu Thr Glu Thr Val Pro
610 615 620
Tyr Glu Ala Phe Lys Phe Ala Asp Phe Gly Val Thr Phe Thr Thr Ala
625 630 635 640
Thr Ala Asn Lys Arg Tyr Thr Phe Gln Phe His Thr Gly Gly Ala Ala
645 650 655
Ile Ile Asp Arg Ile Glu Phe Val Pro Ile Glu Gly Ser Leu Phe Glu
660 665 670
Tyr Glu Thr Lys Gln Gln Leu Glu Lys Ala Arg Lys Ala Val Asn His
675 680 685
Leu Phe Thr Asp Gly Ser Lys Lys Ala Leu Lys Glu Gly Thr Thr Asp
690 695 700
Tyr Glu Ile Asp Gln Ala Ala Asn Val Val Asp Cys Ile Ser Asp Glu
705 710 715 720
Cys Gly His Glu Lys Met Ile Leu Leu Asp Glu Val Lys Tyr Ala Lys
725 730 735
Gln Leu Ser Gln Ala Arg Asn Leu Leu Leu Asn Gly Asn Phe Asp Asp
740 745 750
Leu Tyr Pro Ala Leu Glu Arg Glu Asn Pro Trp Lys Thr Ser Pro His
755 760 765
Val Thr Ile Arg Gln Asp Asn Pro Ile Phe Lys Gly His Tyr Leu Ser
770 775 780
Met Ala Gly Ala Asn Asp Ile Glu Ala Thr Asn Asp Thr Phe Pro Thr
785 790 795 800
Tyr Val Tyr Gln Lys Ile Asp Glu Ala Lys Leu Lys Pro Tyr Thr Arg
805 810 815
Tyr Lys Val Arg Gly Phe Val Gly Ser Ser Lys Ala Leu Glu Leu Leu
820 825 830
Val Thr Arg Tyr Asn Glu Glu Val Asp Ala Ile Leu Asp Val Pro Asp
835 840 845
Asn Ile Pro His Ala Pro Thr Pro Val Cys Gly Glu Phe Asp Arg Cys
850 855 860
Lys Pro Tyr Ser Tyr Pro Pro Leu Leu Pro Glu Cys Asn Pro Glu Phe
865 870 875 880
Ile Asn Gln Met Gln Pro Ser Ser Cys His His Asn Gln Met Val Asp
885 890 895
Tyr Asn Asn Arg Lys His Arg Lys Cys His Gln Ala His Gln Phe Glu
900 905 910
Phe His Ile Asp Thr Gly Thr Ile Asp Leu Val Glu Asp Leu Gly Ile
915 920 925
Trp Val Ile Phe Lys Ile Cys Ala Thr Asp Gly Tyr Ala Ser Leu Asp
930 935 940
Asp Leu Glu Val Ile Glu Glu Gly Ala Leu Gly Val Glu Ala Leu Glu
945 950 955 960
Leu Val Lys Lys Arg Glu Lys Lys Trp Arg His Gln Lys Glu Gln His
965 970 975
Cys Ser Gln Thr Lys His Lys Tyr Asp Ala Ala Lys His Ala Val Met
980 985 990
Ala Leu Phe Thr Asn Thr Arg Tyr Glu Lys Leu Lys Phe Glu Thr Thr
995 1000 1005
Ile Ser Asn Ile Leu Tyr Ala Asp His Leu Val Gln Ser Ile Pro Tyr
1010 1015 1020
Val Tyr Asn Lys Tyr Val Pro Glu Val Pro Gly Met Asn Tyr Glu Leu
1025 1030 1035 1040
Tyr Thr Glu Leu Asn Thr Leu Val Gln Asn Ala Phe Tyr Leu Tyr Asp
1045 1050 1055
Gln Arg Asn Leu Ile Lys Asn Gly Arg Phe Ser Asn Gly Leu Met Tyr
1060 1065 1070
Trp Gln Ala Thr Pro His Ala Arg Val Glu Gln Glu Tyr Glu Lys Ser
1075 1080 1085
Val Leu Val Leu Pro Asn Trp Asp Ala Asn Val Ser Gln Asp Leu Cys
1090 1095 1100
Ile Glu His Asn Arg Gly Tyr Val Leu Arg Val Thr Ala Arg Lys Glu
1105 1110 1115 1120
Asp Pro Gly Ala Gly Asn Val Thr Phe Ser Asp Cys Ala Asn His Val
1125 1130 1135
Asp Lys Leu Ser Phe Thr Ser Cys Asp Ile Ala Thr Asn Ala Val Pro
1140 1145 1150
Gly Ala Gln Ala Asn Asp Pro Ala Ala Gly Val Ala Tyr Gly Gln Gln
1155 1160 1165
Gly Cys Gln Ile Asp Arg Val Pro Tyr Gly Gln Ser Gly Tyr Arg Thr
1170 1175 1180
Asp Gly Thr Asn Gly Met Pro Tyr Gly Gln Ser Gly Asn Arg Ala Asp
1185 1190 1195 1200
Gly Val Pro Tyr Arg Gln Ser Gly Tyr Gly Thr Asp Gly Val Ala His
1205 1210 1215
Asp Gln Pro Gly Tyr Arg Ala Asp Gly Val Ala Tyr Glu Gln Ser Gly
1220 1225 1230
Tyr Arg Ala Asp Gly Val Thr Tyr Asp Gln Ser Ala Asn Gln Thr Arg
1235 1240 1245
Lys Tyr His Gly Cys His Thr Val Gly Leu Pro His Pro Glu His Gly
1250 1255 1260
Cys Cys Tyr Pro Asp Arg Val Ser Asp Gly Gln Gln Leu Ala Tyr Val
1265 1270 1275 1280
Thr Lys Ser Ile Asp Leu Phe Pro Asp Thr Asp Lys Val Arg Ile Asp
1285 1290 1295
Ile Gly Glu Thr Glu Gly Asn Phe Arg Val Glu Ser Val Glu Leu Ile
1300 1305 1310
Cys Met Glu Lys
1315
210> SEQ ID NO 5
<211> LENGTH: 4188
<212> TYPE: DNA
<213> ORGANISM: Bacillus popilliae
<220> FEATURE:
<221> NAME/KEY: CDS
<222> LOCATION: (1)..(4158)
<400> SEQUENCE: 5
atg ttg tgg cta aat act cat tgt tca tta ata gaa ata act ggg agg 48
Met Leu Trp Leu Asn Thr His Cys Ser Leu Ile Glu Ile Thr Gly Arg
1 5 10 15
caa act atg aat caa tat cat aac caa aac gat aac aaa agt tac aac 96
Gln Thr Met Asn Gln Tyr His Asn Gln Asn Asp Asn Lys Ser Tyr Asn
20 25 30
caa agt gga aat gaa gtg caa att ata caa cct tca agt aac gct tta 144
Gln Ser Gly Asn Glu Val Gln Ile Ile Gln Pro Ser Ser Asn Ala Leu
35 40 45
ctt tac agt ccc aac aag tat ccg tat gct acg gat ccc aat gtc ata 192
Leu Tyr Ser Pro Asn Lys Tyr Pro Tyr Ala Thr Asp Pro Asn Val Ile
50 55 60
gca gag ggt aga agt tat aaa aat tgg ttg gat atg tgt gta ggt gta 240
Ala Glu Gly Arg Ser Tyr Lys Asn Trp Leu Asp Met Cys Val Gly Val
65 70 75 80
ggt gac gat aca cga agt ccc gaa gct cgg gtt act gca caa agt tct 288
Gly Asp Asp Thr Arg Ser Pro Glu Ala Arg Val Thr Ala Gln Ser Ser
85 90 95
att tcg act tct ctt ggt ata acc agc aca atc att ggc gct cta ggt 336
Ile Ser Thr Ser Leu Gly Ile Thr Ser Thr Ile Ile Gly Ala Leu Gly
100 105 110
att ccg gtt gta ggc gaa gcc att gga att ttc ggc gca cta ttg gat 384
Ile Pro Val Val Gly Glu Ala Ile Gly Ile Phe Gly Ala Leu Leu Asp
115 120 125
tgg ttg tgg cct gca ggc gct gat cca tgg gta ata ttt atg aat cat 432
Trp Leu Trp Pro Ala Gly Ala Asp Pro Trp Val Ile Phe Met Asn His
130 135 140
gta gaa gaa ctc ata aat tca aaa ata aca gag act gta aaa aat gag 480
Val Glu Glu Leu Ile Asn Ser Lys Ile Thr Glu Thr Val Lys Asn Glu
145 150 155 160
gca att aca aga tta gac ggt tta ggc aat gtc cta gcg cta tat caa 528
Ala Ile Thr Arg Leu Asp Gly Leu Gly Asn Val Leu Ala Leu Tyr Gln
165 170 175
aag gct ttc gaa gaa tgg caa caa cat cct act ctt gaa tca gct cga 576
Lys Ala Phe Glu Glu Trp Gln Gln His Pro Thr Leu Glu Ser Ala Arg
180 185 190
cta cgt gta aca gat gat ttt tct aat gta aat aaa ttt ttt gag gca 624
Leu Arg Val Thr Asp Asp Phe Ser Asn Val Asn Lys Phe Phe Glu Ala
195 200 205
ttt atg cct tca ttc aga gta cca ggt tat gaa gta cca tta tta agc 672
Phe Met Pro Ser Phe Arg Val Pro Gly Tyr Glu Val Pro Leu Leu Ser
210 215 220
gta tac gta tcc gct gca aac ctc cat tta tta tta tta aga gat agc 720
Val Tyr Val Ser Ala Ala Asn Leu His Leu Leu Leu Leu Arg Asp Ser
225 230 235 240
tcg att ttc ggt ttg gat tgg gga tta agt caa act cat gtt aac gat 768
Ser Ile Phe Gly Leu Asp Trp Gly Leu Ser Gln Thr His Val Asn Asp
245 250 255
aat tat aat ctc caa ata agg cgc tct gca gat tat gca aat cat tgt 816
Asn Tyr Asn Leu Gln Ile Arg Arg Ser Ala Asp Tyr Ala Asn His Cys
260 265 270
aca act tgg tat cgg acg ggt tta caa aga ttg caa ggc acc aat gct 864
Thr Thr Trp Tyr Arg Thr Gly Leu Gln Arg Leu Gln Gly Thr Asn Ala
275 280 285
agc agt tgg gtc aat tat aat cga ttt aga aga gaa atg aca cta act 912
Ser Ser Trp Val Asn Tyr Asn Arg Phe Arg Arg Glu Met Thr Leu Thr
290 295 300
gta tta gat gtt tgt gca tta ttt tca agt tat gat tat cgt agt tac 960
Val Leu Asp Val Cys Ala Leu Phe Ser Ser Tyr Asp Tyr Arg Ser Tyr
305 310 315 320
cca atg gag cta agg gga gag ctt acg aga gaa att tat acg gat cca 1008
Pro Met Glu Leu Arg Gly Glu Leu Thr Arg Glu Ile Tyr Thr Asp Pro
325 330 335
gta gga gcc tct ttt tgg gtg aat aga gca cca aac ttc gca tca ata 1056
Val Gly Ala Ser Phe Trp Val Asn Arg Ala Pro Asn Phe Ala Ser Ile
340 345 350
gaa aat aca gta gtt agg caa cca cac ccc ttt act tgg cta gtt act 1104
Glu Asn Thr Val Val Arg Gln Pro His Pro Phe Thr Trp Leu Val Thr
355 360 365
tta aca gtt aat aca ggt caa gtg aga tct ggc gat gga aat tct aac 1152
Leu Thr Val Asn Thr Gly Gln Val Arg Ser Gly Asp Gly Asn Ser Asn
370 375 380
tat tat tgg aaa tca cat agt caa acc gtg agt gaa acc gga ggg tca 1200
Tyr Tyr Trp Lys Ser His Ser Gln Thr Val Ser Glu Thr Gly Gly Ser
385 390 395 400
ggt cct att cag agt cct acc tgt gga agt act ggt aca att tat cgc 1248
Gly Pro Ile Gln Ser Pro Thr Cys Gly Ser Thr Gly Thr Ile Tyr Arg
405 410 415
acg gat aat tta ctt ttt aat cca ttt tta tta ggt gat att tat acc 1296
Thr Asp Asn Leu Leu Phe Asn Pro Phe Leu Leu Gly Asp Ile Tyr Thr
420 425 430
att aat aca ggt tat gtt tct tat ctg gct aat ttg ttt gga atc tat 1344
Ile Asn Thr Gly Tyr Val Ser Tyr Leu Ala Asn Leu Phe Gly Ile Tyr
435 440 445
tca gct aga ttt acg acg act cgt tct att gag ctt ctg tat gag aac 1392
Ser Ala Arg Phe Thr Thr Thr Arg Ser Ile Glu Leu Leu Tyr Glu Asn
450 455 460
caa aga gtt ttc cca gcc tac aat cat caa att cgt gaa tta cct gga 1440
Gln Arg Val Phe Pro Ala Tyr Asn His Gln Ile Arg Glu Leu Pro Gly
465 470 475 480
gta aac tcg gat agg cca act gct gcc gac tat agt cat aga cta tcg 1488
Val Asn Ser Asp Arg Pro Thr Ala Ala Asp Tyr Ser His Arg Leu Ser
485 490 495
tat atc tca ggt ttt gca act gat gtg gga gga acg gtt cta gtt tat 1536
Tyr Ile Ser Gly Phe Ala Thr Asp Val Gly Gly Thr Val Leu Val Tyr
500 505 510
ggg tgg aca tct tca act gct act cgt gag aat aat att acg cta gac 1584
Gly Trp Thr Ser Ser Thr Ala Thr Arg Glu Asn Asn Ile Thr Leu Asp
515 520 525
gac aga ata gta caa ctt cca gct gtt aag gga aca agt ctc aac aat 1632
Asp Arg Ile Val Gln Leu Pro Ala Val Lys Gly Thr Ser Leu Asn Asn
530 535 540
tgc caa gta gtt aga gga act gga ttt aca gga gga gac tgg ttg aag 1680
Cys Gln Val Val Arg Gly Thr Gly Phe Thr Gly Gly Asp Trp Leu Lys
545 550 555 560
cct aat aat aat ggt aca ttt tct cta gct ctt ggt ttc agg tcg act 1728
Pro Asn Asn Asn Gly Thr Phe Ser Leu Ala Leu Gly Phe Arg Ser Thr
565 570 575
tac act tac cgc ctc cgc att cgt tat gct gcc gca gca ggc gga tca 1776
Tyr Thr Tyr Arg Leu Arg Ile Arg Tyr Ala Ala Ala Ala Gly Gly Ser
580 585 590
ggt ttt tct ctt gtt ata tcg gat caa tat gga gaa ttt cca acc aca 1824
Gly Phe Ser Leu Val Ile Ser Asp Gln Tyr Gly Glu Phe Pro Thr Thr
595 600 605
aca gta tcg ctt tcc tcc aca atg tac tca ctg ccc caa aat gta cca 1872
Thr Val Ser Leu Ser Ser Thr Met Tyr Ser Leu Pro Gln Asn Val Pro
610 615 620
tac gag gct ttt aag att gta gat tta cct tct act gtt act att aga 1920
Tyr Glu Ala Phe Lys Ile Val Asp Leu Pro Ser Thr Val Thr Ile Arg
625 630 635 640
aat act tct cct gct tca aca act ttt cga ctt gat ttc cgt ttc att 1968
Asn Thr Ser Pro Ala Ser Thr Thr Phe Arg Leu Asp Phe Arg Phe Ile
645 650 655
gtg cca tta gga ata ctc gca aat ata tta att gac cga att gaa ttt 2016
Val Pro Leu Gly Ile Leu Ala Asn Ile Leu Ile Asp Arg Ile Glu Phe
660 665 670
gtt ccc att gag ggt tcc ttg ttc gag tac gaa acc aaa cag cag cta 2064
Val Pro Ile Glu Gly Ser Leu Phe Glu Tyr Glu Thr Lys Gln Gln Leu
675 680 685
gaa aaa gca agg aaa gcg gtg aac cat ttg ttt aca gat gga tcg aaa 2112
Glu Lys Ala Arg Lys Ala Val Asn His Leu Phe Thr Asp Gly Ser Lys
690 695 700
aag gcg cta aaa gaa ggc acg aca gat tat gag atc gat caa gcc gcc 2160
Lys Ala Leu Lys Glu Gly Thr Thr Asp Tyr Glu Ile Asp Gln Ala Ala
705 710 715 720
aac gtg gtg gat tgt ata tcg gat gag tgt gga cat gag aaa atg atc 2208
Asn Val Val Asp Cys Ile Ser Asp Glu Cys Gly His Glu Lys Met Ile
725 730 735
ctg ttg gat gaa gtg aaa tat gca aaa caa ctc agc caa gcc cgc aat 2256
Leu Leu Asp Glu Val Lys Tyr Ala Lys Gln Leu Ser Gln Ala Arg Asn
740 745 750
tta ctg ctc aat ggg aat ttc gat gat cta tat cca gct ctg gag agg 2304
Leu Leu Leu Asn Gly Asn Phe Asp Asp Leu Tyr Pro Ala Leu Glu Arg
755 760 765
gag aat cca tgg aaa aca agc ccg aat gtt acg atc cgt caa gat aac 2352
Glu Asn Pro Trp Lys Thr Ser Pro Asn Val Thr Ile Arg Gln Asp Asn
770 775 780
ccg att ttt aaa ggc cat tat ctc agt atg gcg ggt gcg aac gat atc 2400
Pro Ile Phe Lys Gly His Tyr Leu Ser Met Ala Gly Ala Asn Asp Ile
785 790 795 800
gag gcc acc aat gat acc ttc ccc acg tat gcc tat caa aaa ata gac 2448
Glu Ala Thr Asn Asp Thr Phe Pro Thr Tyr Ala Tyr Gln Lys Ile Asp
805 810 815
gaa gcc aaa tta aag ccg tat aca cgt tat aaa gtg cgc ggg ttt gtt 2496
Glu Ala Lys Leu Lys Pro Tyr Thr Arg Tyr Lys Val Arg Gly Phe Val
820 825 830
ggc agc agc aaa gct cta gag ctg ttg gtt aca cgc tat aat gaa gaa 2544
Gly Ser Ser Lys Ala Leu Glu Leu Leu Val Thr Arg Tyr Asn Glu Glu
835 840 845
gtc gat gcg att tta gat gta ccg gat aat atc ccg cat gcg ccg ata 2592
Val Asp Ala Ile Leu Asp Val Pro Asp Asn Ile Pro His Ala Pro Ile
850 855 860
cct gtc tgc ggt gaa ttt gat cga tgc aag ccc tat tcg tat cca cct 2640
Pro Val Cys Gly Glu Phe Asp Arg Cys Lys Pro Tyr Ser Tyr Pro Pro
865 870 875 880
tta ctt cca gaa tgt aac cct gag ttt ata aat cag atg caa cca tcc 2688
Leu Leu Pro Glu Cys Asn Pro Glu Phe Ile Asn Gln Met Gln Pro Ser
885 890 895
tct tgc cac cac act cag atg gtc gat tac aat aac atg aac atg agc 2736
Ser Cys His His Thr Gln Met Val Asp Tyr Asn Asn Met Asn Met Ser
900 905 910
acg agt act acc atg aat cct acc ctt acg cct gaa ata gca tcc agc 2784
Thr Ser Thr Thr Met Asn Pro Thr Leu Thr Pro Glu Ile Ala Ser Ser
915 920 925
caa agt gga ttc ggc aga aaa cat cgc aaa tgt cat caa gcg cat caa 2832
Gln Ser Gly Phe Gly Arg Lys His Arg Lys Cys His Gln Ala His Gln
930 935 940
ttt gag ttc cat att gat acc ggg aca atc gat ctg gtc gaa gat ttg 2880
Phe Glu Phe His Ile Asp Thr Gly Thr Ile Asp Leu Val Glu Asp Leu
945 950 955 960
ggc att tgg gtg atc ttc aaa atc tgt gcc aca gat ggt tac gca agc 2928
Gly Ile Trp Val Ile Phe Lys Ile Cys Ala Thr Asp Gly Tyr Ala Ser
965 970 975
tta gat gat ttg gaa gtg att gaa gaa gga gcg ctg ggt gtc gaa gca 2976
Leu Asp Asp Leu Glu Val Ile Glu Glu Gly Ala Leu Gly Val Glu Ala
980 985 990
tta gaa ctt gtc aag aaa aga gaa aag aaa tgg aga cat cag aag gag 3024
Leu Glu Leu Val Lys Lys Arg Glu Lys Lys Trp Arg His Gln Lys Glu
995 1000 1005
cag cac tgt tcg caa acg aaa cac aaa tat gat gcg gcc aaa cat gcg 3072
Gln His Cys Ser Gln Thr Lys His Lys Tyr Asp Ala Ala Lys His Ala
1010 1015 1020
gtg atg gcg tta ttt aca aac acg cgc tat gaa aaa ttg aag ttc gaa 3120
Val Met Ala Leu Phe Thr Asn Thr Arg Tyr Glu Lys Leu Lys Phe Glu
1025 1030 1035 1040
aca acc att tct gac att ttg tat gct gat cat ctc gtg cag tcg atc 3168
Thr Thr Ile Ser Asp Ile Leu Tyr Ala Asp His Leu Val Gln Ser Ile
1045 1050 1055
cct tat gta tat aat aaa tat gta ccg gaa gtt cca ggt atg aat tac 3216
Pro Tyr Val Tyr Asn Lys Tyr Val Pro Glu Val Pro Gly Met Asn Tyr
1060 1065 1070
gaa ctc tat tca gag cta aac aca ctg gtt cag aat gcg ttc tac ctg 3264
Glu Leu Tyr Ser Glu Leu Asn Thr Leu Val Gln Asn Ala Phe Tyr Leu
1075 1080 1085
tat gac cag cgg aat ctg att aaa aat ggg cgc ttt agc aat ggg ctt 3312
Tyr Asp Gln Arg Asn Leu Ile Lys Asn Gly Arg Phe Ser Asn Gly Leu
1090 1095 1100
atg cat tgg caa gct act cct cat gca aga gta gag caa gaa cat gag 3360
Met His Trp Gln Ala Thr Pro His Ala Arg Val Glu Gln Glu His Glu
1105 1110 1115 1120
aaa tcg gtg ctc gtg ctg cca aat tgg gat gcc aat gtg tcg caa gat 3408
Lys Ser Val Leu Val Leu Pro Asn Trp Asp Ala Asn Val Ser Gln Asp
1125 1130 1135
ctt tgt atc gaa cac aat cgc ggt tat gta ttg cgt gtc acg gcg aga 3456
Leu Cys Ile Glu His Asn Arg Gly Tyr Val Leu Arg Val Thr Ala Arg
1140 1145 1150
aaa gaa gat ccg gga gcc ggc aat gtt acc ttt agt gac tgt gca aat 3504
Lys Glu Asp Pro Gly Ala Gly Asn Val Thr Phe Ser Asp Cys Ala Asn
1155 1160 1165
cat gtc aac aag ctg agc ttt act tct tgc gat ata gct aca aac gca 3552
His Val Asn Lys Leu Ser Phe Thr Ser Cys Asp Ile Ala Thr Asn Ala
1170 1175 1180
gtg cca ggt gcc caa gcg aat gat cca gcc gcc gga gta gcc tat gga 3600
Val Pro Gly Ala Gln Ala Asn Asp Pro Ala Ala Gly Val Ala Tyr Gly
1185 1190 1195 1200
caa cag ggt tgt caa ata gat aga gtg ccg tac ggg caa tct gga tat 3648
Gln Gln Gly Cys Gln Ile Asp Arg Val Pro Tyr Gly Gln Ser Gly Tyr
1205 1210 1215
aga aca gac gga aca aat gga atg ccg tac ggg cag tct ggt aat cga 3696
Arg Thr Asp Gly Thr Asn Gly Met Pro Tyr Gly Gln Ser Gly Asn Arg
1220 1225 1230
gcg gac ggg gtg cca tac aga caa tcc gga tat gga aca gat gga gta 3744
Ala Asp Gly Val Pro Tyr Arg Gln Ser Gly Tyr Gly Thr Asp Gly Val
1235 1240 1245
gcg cac gac caa cct gga tat cga gca gac gga gca gcg tac gaa cag 3792
Ala His Asp Gln Pro Gly Tyr Arg Ala Asp Gly Ala Ala Tyr Glu Gln
1250 1255 1260
tct ggt cat cga gca gac gga gta gcg tac gaa caa tct gga tat cga 3840
Ser Gly His Arg Ala Asp Gly Val Ala Tyr Glu Gln Ser Gly Tyr Arg
1265 1270 1275 1280
gca ggt gga gta gcg tac gaa cag tct ggt cat cga gca gat gga gtg 3888
Ala Gly Gly Val Ala Tyr Glu Gln Ser Gly His Arg Ala Asp Gly Val
1285 1290 1295
ccg tac gga caa tct gga tat gga aca gac gga gta acg tac gac caa 3936
Pro Tyr Gly Gln Ser Gly Tyr Gly Thr Asp Gly Val Thr Tyr Asp Gln
1300 1305 1310
tct gtc aaa caa acc cgc aaa tac cat ggt tgc cat aca gac ggg ctg 3984
Ser Val Lys Gln Thr Arg Lys Tyr His Gly Cys His Thr Asp Gly Leu
1315 1320 1325
cca cat cca gag cat ggt tgt tgt tat cca gac aga gta agc gat ggc 4032
Pro His Pro Glu His Gly Cys Cys Tyr Pro Asp Arg Val Ser Asp Gly
1330 1335 1340
caa cag ctt gca tat gta aca aaa tcg att gat ctg ttc ccg gat aca 4080
Gln Gln Leu Ala Tyr Val Thr Lys Ser Ile Asp Leu Phe Pro Asp Thr
1345 1350 1355 1360
gat aaa gtc cgg atc gac att gga gaa acc gaa ggg aac ttt aga gtg 4128
Asp Lys Val Arg Ile Asp Ile Gly Glu Thr Glu Gly Asn Phe Arg Val
1365 1370 1375
gaa agt gtg gaa ttg att tgt atg gaa aag taaatcatca caagtaaaag 4178
Glu Ser Val Glu Leu Ile Cys Met Glu Lys
1380 1385
tatcgtttac 4188
<210> SEQ ID NO 6
<211> LENGTH: 1386
<212> TYPE: PRT
<213> ORGANISM: Bacillus popilliae
<400> SEQUENCE: 6
Met Leu Trp Leu Asn Thr His Cys Ser Leu Ile Glu Ile Thr Gly Arg
1 5 10 15
Gln Thr Met Asn Gln Tyr His Asn Gln Asn Asp Asn Lys Ser Tyr Asn
20 25 30
Gln Ser Gly Asn Glu Val Gln Ile Ile Gln Pro Ser Ser Asn Ala Leu
35 40 45
Leu Tyr Ser Pro Asn Lys Tyr Pro Tyr Ala Thr Asp Pro Asn Val Ile
50 55 60
Ala Glu Gly Arg Ser Tyr Lys Asn Trp Leu Asp Met Cys Val Gly Val
65 70 75 80
Gly Asp Asp Thr Arg Ser Pro Glu Ala Arg Val Thr Ala Gln Ser Ser
85 90 95
Ile Ser Thr Ser Leu Gly Ile Thr Ser Thr Ile Ile Gly Ala Leu Gly
100 105 110
Ile Pro Val Val Gly Glu Ala Ile Gly Ile Phe Gly Ala Leu Leu Asp
115 120 125
Trp Leu Trp Pro Ala Gly Ala Asp Pro Trp Val Ile Phe Met Asn His
130 135 140
Val Glu Glu Leu Ile Asn Ser Lys Ile Thr Glu Thr Val Lys Asn Glu
145 150 155 160
Ala Ile Thr Arg Leu Asp Gly Leu Gly Asn Val Leu Ala Leu Tyr Gln
165 170 175
Lys Ala Phe Glu Glu Trp Gln Gln His Pro Thr Leu Glu Ser Ala Arg
180 185 190
Leu Arg Val Thr Asp Asp Phe Ser Asn Val Asn Lys Phe Phe Glu Ala
195 200 205
Phe Met Pro Ser Phe Arg Val Pro Gly Tyr Glu Val Pro Leu Leu Ser
210 215 220
Val Tyr Val Ser Ala Ala Asn Leu His Leu Leu Leu Leu Arg Asp Ser
225 230 235 240
Ser Ile Phe Gly Leu Asp Trp Gly Leu Ser Gln Thr His Val Asn Asp
245 250 255
Asn Tyr Asn Leu Gln Ile Arg Arg Ser Ala Asp Tyr Ala Asn His Cys
260 265 270
Thr Thr Trp Tyr Arg Thr Gly Leu Gln Arg Leu Gln Gly Thr Asn Ala
275 280 285
Ser Ser Trp Val Asn Tyr Asn Arg Phe Arg Arg Glu Met Thr Leu Thr
290 295 300
Val Leu Asp Val Cys Ala Leu Phe Ser Ser Tyr Asp Tyr Arg Ser Tyr
305 310 315 320
Pro Met Glu Leu Arg Gly Glu Leu Thr Arg Glu Ile Tyr Thr Asp Pro
325 330 335
Val Gly Ala Ser Phe Trp Val Asn Arg Ala Pro Asn Phe Ala Ser Ile
340 345 350
Glu Asn Thr Val Val Arg Gln Pro His Pro Phe Thr Trp Leu Val Thr
355 360 365
Leu Thr Val Asn Thr Gly Gln Val Arg Ser Gly Asp Gly Asn Ser Asn
370 375 380
Tyr Tyr Trp Lys Ser His Ser Gln Thr Val Ser Glu Thr Gly Gly Ser
385 390 395 400
Gly Pro Ile Gln Ser Pro Thr Cys Gly Ser Thr Gly Thr Ile Tyr Arg
405 410 415
Thr Asp Asn Leu Leu Phe Asn Pro Phe Leu Leu Gly Asp Ile Tyr Thr
420 425 430
Ile Asn Thr Gly Tyr Val Ser Tyr Leu Ala Asn Leu Phe Gly Ile Tyr
435 440 445
Ser Ala Arg Phe Thr Thr Thr Arg Ser Ile Glu Leu Leu Tyr Glu Asn
450 455 460
Gln Arg Val Phe Pro Ala Tyr Asn His Gln Ile Arg Glu Leu Pro Gly
465 470 475 480
Val Asn Ser Asp Arg Pro Thr Ala Ala Asp Tyr Ser His Arg Leu Ser
485 490 495
Tyr Ile Ser Gly Phe Ala Thr Asp Val Gly Gly Thr Val Leu Val Tyr
500 505 510
Gly Trp Thr Ser Ser Thr Ala Thr Arg Glu Asn Asn Ile Thr Leu Asp
515 520 525
Asp Arg Ile Val Gln Leu Pro Ala Val Lys Gly Thr Ser Leu Asn Asn
530 535 540
Cys Gln Val Val Arg Gly Thr Gly Phe Thr Gly Gly Asp Trp Leu Lys
545 550 555 560
Pro Asn Asn Asn Gly Thr Phe Ser Leu Ala Leu Gly Phe Arg Ser Thr
565 570 575
Tyr Thr Tyr Arg Leu Arg Ile Arg Tyr Ala Ala Ala Ala Gly Gly Ser
580 585 590
Gly Phe Ser Leu Val Ile Ser Asp Gln Tyr Gly Glu Phe Pro Thr Thr
595 600 605
Thr Val Ser Leu Ser Ser Thr Met Tyr Ser Leu Pro Gln Asn Val Pro
610 615 620
Tyr Glu Ala Phe Lys Ile Val Asp Leu Pro Ser Thr Val Thr Ile Arg
625 630 635 640
Asn Thr Ser Pro Ala Ser Thr Thr Phe Arg Leu Asp Phe Arg Phe Ile
645 650 655
Val Pro Leu Gly Ile Leu Ala Asn Ile Leu Ile Asp Arg Ile Glu Phe
660 665 670
Val Pro Ile Glu Gly Ser Leu Phe Glu Tyr Glu Thr Lys Gln Gln Leu
675 680 685
Glu Lys Ala Arg Lys Ala Val Asn His Leu Phe Thr Asp Gly Ser Lys
690 695 700
Lys Ala Leu Lys Glu Gly Thr Thr Asp Tyr Glu Ile Asp Gln Ala Ala
705 710 715 720
Asn Val Val Asp Cys Ile Ser Asp Glu Cys Gly His Glu Lys Met Ile
725 730 735
Leu Leu Asp Glu Val Lys Tyr Ala Lys Gln Leu Ser Gln Ala Arg Asn
740 745 750
Leu Leu Leu Asn Gly Asn Phe Asp Asp Leu Tyr Pro Ala Leu Glu Arg
755 760 765
Glu Asn Pro Trp Lys Thr Ser Pro Asn Val Thr Ile Arg Gln Asp Asn
770 775 780
Pro Ile Phe Lys Gly His Tyr Leu Ser Met Ala Gly Ala Asn Asp Ile
785 790 795 800
Glu Ala Thr Asn Asp Thr Phe Pro Thr Tyr Ala Tyr Gln Lys Ile Asp
805 810 815
Glu Ala Lys Leu Lys Pro Tyr Thr Arg Tyr Lys Val Arg Gly Phe Val
820 825 830
Gly Ser Ser Lys Ala Leu Glu Leu Leu Val Thr Arg Tyr Asn Glu Glu
835 840 845
Val Asp Ala Ile Leu Asp Val Pro Asp Asn Ile Pro His Ala Pro Ile
850 855 860
Pro Val Cys Gly Glu Phe Asp Arg Cys Lys Pro Tyr Ser Tyr Pro Pro
865 870 875 880
Leu Leu Pro Glu Cys Asn Pro Glu Phe Ile Asn Gln Met Gln Pro Ser
885 890 895
Ser Cys His His Thr Gln Met Val Asp Tyr Asn Asn Met Asn Met Ser
900 905 910
Thr Ser Thr Thr Met Asn Pro Thr Leu Thr Pro Glu Ile Ala Ser Ser
915 920 925
Gln Ser Gly Phe Gly Arg Lys His Arg Lys Cys His Gln Ala His Gln
930 935 940
Phe Glu Phe His Ile Asp Thr Gly Thr Ile Asp Leu Val Glu Asp Leu
945 950 955 960
Gly Ile Trp Val Ile Phe Lys Ile Cys Ala Thr Asp Gly Tyr Ala Ser
965 970 975
Leu Asp Asp Leu Glu Val Ile Glu Glu Gly Ala Leu Gly Val Glu Ala
980 985 990
Leu Glu Leu Val Lys Lys Arg Glu Lys Lys Trp Arg His Gln Lys Glu
995 1000 1005
Gln His Cys Ser Gln Thr Lys His Lys Tyr Asp Ala Ala Lys His Ala
1010 1015 1020
Val Met Ala Leu Phe Thr Asn Thr Arg Tyr Glu Lys Leu Lys Phe Glu
1025 1030 1035 1040
Thr Thr Ile Ser Asp Ile Leu Tyr Ala Asp His Leu Val Gln Ser Ile
1045 1050 1055
Pro Tyr Val Tyr Asn Lys Tyr Val Pro Glu Val Pro Gly Met Asn Tyr
1060 1065 1070
Glu Leu Tyr Ser Glu Leu Asn Thr Leu Val Gln Asn Ala Phe Tyr Leu
1075 1080 1085
Tyr Asp Gln Arg Asn Leu Ile Lys Asn Gly Arg Phe Ser Asn Gly Leu
1090 1095 1100
Met His Trp Gln Ala Thr Pro His Ala Arg Val Glu Gln Glu His Glu
1105 1110 1115 1120
Lys Ser Val Leu Val Leu Pro Asn Trp Asp Ala Asn Val Ser Gln Asp
1125 1130 1135
Leu Cys Ile Glu His Asn Arg Gly Tyr Val Leu Arg Val Thr Ala Arg
1140 1145 1150
Lys Glu Asp Pro Gly Ala Gly Asn Val Thr Phe Ser Asp Cys Ala Asn
1155 1160 1165
His Val Asn Lys Leu Ser Phe Thr Ser Cys Asp Ile Ala Thr Asn Ala
1170 1175 1180
Val Pro Gly Ala Gln Ala Asn Asp Pro Ala Ala Gly Val Ala Tyr Gly
1185 1190 1195 1200
Gln Gln Gly Cys Gln Ile Asp Arg Val Pro Tyr Gly Gln Ser Gly Tyr
1205 1210 1215
Arg Thr Asp Gly Thr Asn Gly Met Pro Tyr Gly Gln Ser Gly Asn Arg
1220 1225 1230
Ala Asp Gly Val Pro Tyr Arg Gln Ser Gly Tyr Gly Thr Asp Gly Val
1235 1240 1245
Ala His Asp Gln Pro Gly Tyr Arg Ala Asp Gly Ala Ala Tyr Glu Gln
1250 1255 1260
Ser Gly His Arg Ala Asp Gly Val Ala Tyr Glu Gln Ser Gly Tyr Arg
1265 1270 1275 1280
Ala Gly Gly Val Ala Tyr Glu Gln Ser Gly His Arg Ala Asp Gly Val
1285 1290 1295
Pro Tyr Gly Gln Ser Gly Tyr Gly Thr Asp Gly Val Thr Tyr Asp Gln
1300 1305 1310
Ser Val Lys Gln Thr Arg Lys Tyr His Gly Cys His Thr Asp Gly Leu
1315 1320 1325
Pro His Pro Glu His Gly Cys Cys Tyr Pro Asp Arg Val Ser Asp Gly
1330 1335 1340
Gln Gln Leu Ala Tyr Val Thr Lys Ser Ile Asp Leu Phe Pro Asp Thr
1345 1350 1355 1360
Asp Lys Val Arg Ile Asp Ile Gly Glu Thr Glu Gly Asn Phe Arg Val
1365 1370 1375
Glu Ser Val Glu Leu Ile Cys Met Glu Lys
1380 1385
<210> SEQ ID NO 7
<211> LENGTH: 795
<212> TYPE: DNA
<213> ORGANISM: Bacillus popilliae
<220> FEATURE:
<221> NAME/KEY: CDS
<222> LOCATION: (1)..(795)
<400> SEQUENCE: 7
caa tat cat aac caa aac gat aac aaa agt tac aac caa agt gga aat 48
Gln Tyr His Asn Gln Asn Asp Asn Lys Ser Tyr Asn Gln Ser Gly Asn
1 5 10 15
gac atg caa att ata caa cct tca agt aac gct tta ctt tac agt ccc 96
Asp Met Gln Ile Ile Gln Pro Ser Ser Asn Ala Leu Leu Tyr Ser Pro
20 25 30
aat aag tat ccg tat gcc acg gat ccc aat gtc ata gca gag ggg aga 144
Asn Lys Tyr Pro Tyr Ala Thr Asp Pro Asn Val Ile Ala Glu Gly Arg
35 40 45
agt tat aat aat tgg ttg gat acg tgt gta ggt gta ggc gat ggt act 192
Ser Tyr Asn Asn Trp Leu Asp Thr Cys Val Gly Val Gly Asp Gly Thr
50 55 60
cga agt ccc gag gct tat gct att gcc gaa gag gct gtt ggt ctt tcg 240
Arg Ser Pro Glu Ala Tyr Ala Ile Ala Glu Glu Ala Val Gly Leu Ser
65 70 75 80
att gat ctt ctc gcc gaa act ata tac ttt cta ggc ttt ccg att gca 288
Ile Asp Leu Leu Ala Glu Thr Ile Tyr Phe Leu Gly Phe Pro Ile Ala
85 90 95
tct cca atc act cgt gca cta tct gct tta ctg ggg ggg cta ttt tct 336
Ser Pro Ile Thr Arg Ala Leu Ser Ala Leu Leu Gly Gly Leu Phe Ser
100 105 110
tct ggg gat acg ctg atg caa cat gta gaa caa ctc ata aat caa aaa 384
Ser Gly Asp Thr Leu Met Gln His Val Glu Gln Leu Ile Asn Gln Lys
115 120 125
ata gaa att tat gca aga aat acg gca ctt gca gaa tta ttg ggt tta 432
Ile Glu Ile Tyr Ala Arg Asn Thr Ala Leu Ala Glu Leu Leu Gly Leu
130 135 140
cgt aat gca tta gaa gtc tat agt gtg gct tta gaa tat tgg caa caa 480
Arg Asn Ala Leu Glu Val Tyr Ser Val Ala Leu Glu Tyr Trp Gln Gln
145 150 155 160
aat cgt aat agt gca caa gct caa gaa agt gta aga tca aga ttc cgt 528
Asn Arg Asn Ser Ala Gln Ala Gln Glu Ser Val Arg Ser Arg Phe Arg
165 170 175
agt cta gaa act ata ttt ata caa agg atg cct tta ttt gca ata caa 576
Ser Leu Glu Thr Ile Phe Ile Gln Arg Met Pro Leu Phe Ala Ile Gln
180 185 190
ggt tat gag gtg ccg tta tta tcc gtg tat gca gcc gct gca aat ctc 624
Gly Tyr Glu Val Pro Leu Leu Ser Val Tyr Ala Ala Ala Ala Asn Leu
195 200 205
cat tta cta tta tta agg gat agc tct att cac ggt tta gac tgg gga 672
His Leu Leu Leu Leu Arg Asp Ser Ser Ile His Gly Leu Asp Trp Gly
210 215 220
ttt gat caa gga gag gtt aac agt aat tac gat cgc caa att agg ctc 720
Phe Asp Gln Gly Glu Val Asn Ser Asn Tyr Asp Arg Gln Ile Arg Leu
225 230 235 240
tct gca gag tat gca aat cat tgt ata act tgg tat cag gcg ggt tta 768
Ser Ala Glu Tyr Ala Asn His Cys Ile Thr Trp Tyr Gln Ala Gly Leu
245 250 255
caa gga ttg cag ggc acc agg ggg cga 795
Gln Gly Leu Gln Gly Thr Arg Gly Arg
260 265
<210> SEQ ID NO 8
<211> LENGTH: 265
<212> TYPE: PRT
<213> ORGANISM: Bacillus popilliae
<400> SEQUENCE: 8
Gln Tyr His Asn Gln Asn Asp Asn Lys Ser Tyr Asn Gln Ser Gly Asn
1 5 10 15
Asp Met Gln Ile Ile Gln Pro Ser Ser Asn Ala Leu Leu Tyr Ser Pro
20 25 30
Asn Lys Tyr Pro Tyr Ala Thr Asp Pro Asn Val Ile Ala Glu Gly Arg
35 40 45
Ser Tyr Asn Asn Trp Leu Asp Thr Cys Val Gly Val Gly Asp Gly Thr
50 55 60
Arg Ser Pro Glu Ala Tyr Ala Ile Ala Glu Glu Ala Val Gly Leu Ser
65 70 75 80
Ile Asp Leu Leu Ala Glu Thr Ile Tyr Phe Leu Gly Phe Pro Ile Ala
85 90 95
Ser Pro Ile Thr Arg Ala Leu Ser Ala Leu Leu Gly Gly Leu Phe Ser
100 105 110
Ser Gly Asp Thr Leu Met Gln His Val Glu Gln Leu Ile Asn Gln Lys
115 120 125
Ile Glu Ile Tyr Ala Arg Asn Thr Ala Leu Ala Glu Leu Leu Gly Leu
130 135 140
Arg Asn Ala Leu Glu Val Tyr Ser Val Ala Leu Glu Tyr Trp Gln Gln
145 150 155 160
Asn Arg Asn Ser Ala Gln Ala Gln Glu Ser Val Arg Ser Arg Phe Arg
165 170 175
Ser Leu Glu Thr Ile Phe Ile Gln Arg Met Pro Leu Phe Ala Ile Gln
180 185 190
Gly Tyr Glu Val Pro Leu Leu Ser Val Tyr Ala Ala Ala Ala Asn Leu
195 200 205
His Leu Leu Leu Leu Arg Asp Ser Ser Ile His Gly Leu Asp Trp Gly
210 215 220
Phe Asp Gln Gly Glu Val Asn Ser Asn Tyr Asp Arg Gln Ile Arg Leu
225 230 235 240
Ser Ala Glu Tyr Ala Asn His Cys Ile Thr Trp Tyr Gln Ala Gly Leu
245 250 255
Gln Gly Leu Gln Gly Thr Arg Gly Arg
260 265
<210> SEQ ID NO 9
<211> LENGTH: 459
<212> TYPE: DNA
<213> ORGANISM: Bacillus popilliae
<220> FEATURE:
<221> NAME/KEY: CDS
<222> LOCATION: (1)..(459)
<400> SEQUENCE: 9
tat gag gtc cca tta tta tcc gtg tat gca gac gct gca aat ctc cat 48
Tyr Glu Val Pro Leu Leu Ser Val Tyr Ala Asp Ala Ala Asn Leu His
1 5 10 15
ttg cta ata tta agg gac agc tat att tac ggt gca ttc tgg ggg ttt 96
Leu Leu Ile Leu Arg Asp Ser Tyr Ile Tyr Gly Ala Phe Trp Gly Phe
20 25 30
gat gaa gat gaa tat tac cgt aat tac gct cgc caa att agg ctc tcc 144
Asp Glu Asp Glu Tyr Tyr Arg Asn Tyr Ala Arg Gln Ile Arg Leu Ser
35 40 45
gca gag tat gca aat cat tgt aca act tgg tat cag acg ggt tta cga 192
Ala Glu Tyr Ala Asn His Cys Thr Thr Trp Tyr Gln Thr Gly Leu Arg
50 55 60
aga ttg cag ggc acc cga gct aca gat tgg atc aat tat aat cga ttt 240
Arg Leu Gln Gly Thr Arg Ala Thr Asp Trp Ile Asn Tyr Asn Arg Phe
65 70 75 80
aga aga gaa ctg aca cta aca gta tta gat att tgt gca tta ttt tct 288
Arg Arg Glu Leu Thr Leu Thr Val Leu Asp Ile Cys Ala Leu Phe Ser
85 90 95
agc tat gat att cct agt tac ccg atg ggg aca aag ata cag ctt acg 336
Ser Tyr Asp Ile Pro Ser Tyr Pro Met Gly Thr Lys Ile Gln Leu Thr
100 105 110
aga gaa att tat acc gat cca gta gta cac tct gac tgg ttg caa tca 384
Arg Glu Ile Tyr Thr Asp Pro Val Val His Ser Asp Trp Leu Gln Ser
115 120 125
acg agt ccg gga tta ata tca ttc tca tca cta gaa aat cta gtc gtt 432
Thr Ser Pro Gly Leu Ile Ser Phe Ser Ser Leu Glu Asn Leu Val Val
130 135 140
cgg gca cca cat ctt ttt act tgg ctt 459
Arg Ala Pro His Leu Phe Thr Trp Leu
145 150
<210> SEQ ID NO 10
<211> LENGTH: 153
<212> TYPE: PRT
<213> ORGANISM: Bacillus popilliae
<400> SEQUENCE: 10
Tyr Glu Val Pro Leu Leu Ser Val Tyr Ala Asp Ala Ala Asn Leu His
1 5 10 15
Leu Leu Ile Leu Arg Asp Ser Tyr Ile Tyr Gly Ala Phe Trp Gly Phe
20 25 30
Asp Glu Asp Glu Tyr Tyr Arg Asn Tyr Ala Arg Gln Ile Arg Leu Ser
35 40 45
Ala Glu Tyr Ala Asn His Cys Thr Thr Trp Tyr Gln Thr Gly Leu Arg
50 55 60
Arg Leu Gln Gly Thr Arg Ala Thr Asp Trp Ile Asn Tyr Asn Arg Phe
65 70 75 80
Arg Arg Glu Leu Thr Leu Thr Val Leu Asp Ile Cys Ala Leu Phe Ser
85 90 95
Ser Tyr Asp Ile Pro Ser Tyr Pro Met Gly Thr Lys Ile Gln Leu Thr
100 105 110
Arg Glu Ile Tyr Thr Asp Pro Val Val His Ser Asp Trp Leu Gln Ser
115 120 125
Thr Ser Pro Gly Leu Ile Ser Phe Ser Ser Leu Glu Asn Leu Val Val
130 135 140
Arg Ala Pro His Leu Phe Thr Trp Leu
145 150
<210> SEQ ID NO 11
<211> LENGTH: 25
<212> TYPE: DNA
<213> ORGANISM: Artificial Sequence
<220> FEATURE:
<223> OTHER INFORMATION: primer
<220> FEATURE:
<221> NAME/KEY: misc_feature
<222> LOCATION: (5)
<223> OTHER INFORMATION: n=a or g or c or t
<220> FEATURE:
<221> NAME/KEY: misc_feature
<222> LOCATION: (11)..(12)
<223> OTHER INFORMATION: n=a or g or c or t
<400> SEQUENCE: 11
atgcngaagt nnatttacac gtgtt 25
<210> SEQ ID NO 12
<211> LENGTH: 17
<212> TYPE: DNA
<213> ORGANISM: Artificial Sequence
<220> FEATURE:
<223> OTHER INFORMATION: primer
<220> FEATURE:
<221> NAME/KEY: misc_feature
<222> LOCATION: (14)..(15)
<223> OTHER INFORMATION: n=a or g or c or t
<400> SEQUENCE: 12
atgttgtggc taannac 17
<210> SEQ ID NO 13
<211> LENGTH: 20
<212> TYPE: DNA
<213> ORGANISM: Artificial Sequence
<220> FEATURE:
<223> OTHER INFORMATION: primer
<400> SEQUENCE: 13
gtaaacgata cttttacttg 20
<210> SEQ ID NO 14
<211> LENGTH: 20
<212> TYPE: DNA
<213> ORGANISM: Artificial Sequence
<220> FEATURE:
<223> OTHER INFORMATION: primer
<400> SEQUENCE: 14
tctcgatcac gaatgatcat 20
<210> SEQ ID NO 15
<211> LENGTH: 20
<212> TYPE: DNA
<213> ORGANISM: Artificial Sequence
<220> FEATURE:
<223> OTHER INFORMATION: primer
<400> SEQUENCE: 15
gtgtcatttc tcttctaaat 20
<210> SEQ ID NO 16
<211> LENGTH: 30
<212> TYPE: DNA
<213> ORGANISM: Artificial Sequence
<220> FEATURE:
<223> OTHER INFORMATION: primer
<400> SEQUENCE: 16
gtggaacaag aatatgagaa atctgtactc 30
<210> SEQ ID NO 17
<211> LENGTH: 4359
<212> TYPE: DNA
<213> ORGANISM: Bacillus popilliae
<220> FEATURE:
<221> NAME/KEY: CDS
<222> LOCATION: (250)..(4245)
<400> SEQUENCE: 17
ttgccagatg ttcatatcga ttgtcaatat gtgacggtat gtgatttaca aatgatgccc 60
gttcatgagg gagcttgtca atttgtgaag attagcggag aatttcaatt ttattcactt 120
taacattagc tacaaatgtt gtggctaagc accccattat ttatcaacgg gtaacaacca 180
ggaggggtaa ctttgaatca gtatcaacat caaaacgata acaaaagtta caatcaagat 240
ggaaatgaa atg cag atc ata caa cct tca agt aac gct tta ctt tac agt 291
Met Gln Ile Ile Gln Pro Ser Ser Asn Ala Leu Leu Tyr Ser
1 5 10
ccc aat aag tat ccg tat gcc acg gat ccc aat gtc ata gca gag ggg 339
Pro Asn Lys Tyr Pro Tyr Ala Thr Asp Pro Asn Val Ile Ala Glu Gly
15 20 25 30
aga agt tat aat aat tgg ttg gat acg tgt gta ggt gta ggc gat ggt 387
Arg Ser Tyr Asn Asn Trp Leu Asp Thr Cys Val Gly Val Gly Asp Gly
35 40 45
act cga agt ccc gag gct tat gct att gcc gaa gag gct gtc ggt ctt 435
Thr Arg Ser Pro Glu Ala Tyr Ala Ile Ala Glu Glu Ala Val Gly Leu
50 55 60
tca att gat ata ttg gcc gaa att ata tac tat cta ggc ttc ccg att 483
Ser Ile Asp Ile Leu Ala Glu Ile Ile Tyr Tyr Leu Gly Phe Pro Ile
65 70 75
gca tct cca ctc act cgt gca cta agt gct ata gcg gga cag cta ttt 531
Ala Ser Pro Leu Thr Arg Ala Leu Ser Ala Ile Ala Gly Gln Leu Phe
80 85 90
tct tct ggg gat acg ctc atg caa cat att gaa caa ctc ata aat caa 579
Ser Ser Gly Asp Thr Leu Met Gln His Ile Glu Gln Leu Ile Asn Gln
95 100 105 110
aaa ata gcg gaa tat gcc aga aat aag gcg ctt gca gaa ttt cag ggt 627
Lys Ile Ala Glu Tyr Ala Arg Asn Lys Ala Leu Ala Glu Phe Gln Gly
115 120 125
tta ggt aga caa tat ggg tta tat tta gag gct tta gaa gat tgg gaa 675
Leu Gly Arg Gln Tyr Gly Leu Tyr Leu Glu Ala Leu Glu Asp Trp Glu
130 135 140
caa aat cgt ctt agt caa cca cat aaa gag cgt gta aga caa aca ttc 723
Gln Asn Arg Leu Ser Gln Pro His Lys Glu Arg Val Arg Gln Thr Phe
145 150 155
cgt att ctt gat aat agc ttt aca tca tct ata cct tca ttt gca gta 771
Arg Ile Leu Asp Asn Ser Phe Thr Ser Ser Ile Pro Ser Phe Ala Val
160 165 170
cga aat tat gag gtt cca tta tta tcc gtg tat gca gac gct gca aat 819
Arg Asn Tyr Glu Val Pro Leu Leu Ser Val Tyr Ala Asp Ala Ala Asn
175 180 185 190
ctc cat ttg cta ata tta agg gac agc tat att tac ggt gca ttc tgg 867
Leu His Leu Leu Ile Leu Arg Asp Ser Tyr Ile Tyr Gly Ala Phe Trp
195 200 205
ggg ttt gat gaa gat gaa tat tac cgt aat tac gct cgc caa att agg 915
Gly Phe Asp Glu Asp Glu Tyr Tyr Arg Asn Tyr Ala Arg Gln Ile Arg
210 215 220
ctc tcc gca gag tat gca aat cat tgt aca act tgg tat cag acg ggt 963
Leu Ser Ala Glu Tyr Ala Asn His Cys Thr Thr Trp Tyr Gln Thr Gly
225 230 235
tta cga aga ttg cag ggc acc cga gct aca gat tgg atc aat tat aat 1011
Leu Arg Arg Leu Gln Gly Thr Arg Ala Thr Asp Trp Ile Asn Tyr Asn
240 245 250
cga ttt aga aga gaa atg aca cta aca gta tta gat att tgt gca tta 1059
Arg Phe Arg Arg Glu Met Thr Leu Thr Val Leu Asp Ile Cys Ala Leu
255 260 265 270
ttt tct agc tat gat att cct agt tac ccg atg ggg aca aag ata cag 1107
Phe Ser Ser Tyr Asp Ile Pro Ser Tyr Pro Met Gly Thr Lys Ile Gln
275 280 285
ctt acg aga gaa att tat acc gat cca gta gta cac tct gac tgg ttg 1155
Leu Thr Arg Glu Ile Tyr Thr Asp Pro Val Val His Ser Asp Trp Leu
290 295 300
caa tca acg agt ccg gga tta ata tca ttc tca tca cta gaa aat cta 1203
Gln Ser Thr Ser Pro Gly Leu Ile Ser Phe Ser Ser Leu Glu Asn Leu
305 310 315
gtc gtt cgg gca cca cat ctt ttt act tgg ctt tct cga gta aca att 1251
Val Val Arg Ala Pro His Leu Phe Thr Trp Leu Ser Arg Val Thr Ile
320 325 330
gat aca ggt ata ttg tct act gta att ggc ggg cag tat agt aat aac 1299
Asp Thr Gly Ile Leu Ser Thr Val Ile Gly Gly Gln Tyr Ser Asn Asn
335 340 345 350
aat ttt tgg cga aca cat tat caa act ttg cgt aca acc ggg ggc aca 1347
Asn Phe Trp Arg Thr His Tyr Gln Thr Leu Arg Thr Thr Gly Gly Thr
355 360 365
tct ttc caa agt cct acc tat ggc tcg aca gcg ttt cca att caa cgc 1395
Ser Phe Gln Ser Pro Thr Tyr Gly Ser Thr Ala Phe Pro Ile Gln Arg
370 375 380
acg aat aca ttg act ttc tcc ggc gat gtt tac acc ata gag tca agt 1443
Thr Asn Thr Leu Thr Phe Ser Gly Asp Val Tyr Thr Ile Glu Ser Ser
385 390 395
gtt gtt acg agg agt tcc ttg tat gga gct aat tcg gtt gca ttt acg 1491
Val Val Thr Arg Ser Ser Leu Tyr Gly Ala Asn Ser Val Ala Phe Thr
400 405 410
ggt act act ggt cgg tca cta tat gag aac cca acg gtt tat ccg ttt 1539
Gly Thr Thr Gly Arg Ser Leu Tyr Glu Asn Pro Thr Val Tyr Pro Phe
415 420 425 430
gca caa aaa tta att cat gaa tta cct gga gta gac tca ggt aga cca 1587
Ala Gln Lys Leu Ile His Glu Leu Pro Gly Val Asp Ser Gly Arg Pro
435 440 445
aat gct acc aac tat agt cat aga ctg tcg tat atc tca ggt ttc agt 1635
Asn Ala Thr Asn Tyr Ser His Arg Leu Ser Tyr Ile Ser Gly Phe Ser
450 455 460
ctt ggt tat tct cct tct gga acg ggt cta gtt tat ggg tgg aca tct 1683
Leu Gly Tyr Ser Pro Ser Gly Thr Gly Leu Val Tyr Gly Trp Thr Ser
465 470 475
aca act gct act cgt gag aat aat att acg cta gac gac aga ata gta 1731
Thr Thr Ala Thr Arg Glu Asn Asn Ile Thr Leu Asp Asp Arg Ile Val
480 485 490
cag ctt cca gct gtt aag gga gca agt ctc aat aat tgc caa gta gta 1779
Gln Leu Pro Ala Val Lys Gly Ala Ser Leu Asn Asn Cys Gln Val Val
495 500 505 510
aaa ggg act gga ttt aca gga gga gac tgg ttg aaa cct aat aat aat 1827
Lys Gly Thr Gly Phe Thr Gly Gly Asp Trp Leu Lys Pro Asn Asn Asn
515 520 525
ggt aca ttt tct atg tac ttt gca ttc agg tcg gct tac act tac cac 1875
Gly Thr Phe Ser Met Tyr Phe Ala Phe Arg Ser Ala Tyr Thr Tyr His
530 535 540
ttc cgc att cgt tat gct tcc tca gca agt ttt tct ttt gtt ata tcg 1923
Phe Arg Ile Arg Tyr Ala Ser Ser Ala Ser Phe Ser Phe Val Ile Ser
545 550 555
gaa gaa tat gga cgt ttt cca acc aca aca gtg ccg ctt ctc tcc aca 1971
Glu Glu Tyr Gly Arg Phe Pro Thr Thr Thr Val Pro Leu Leu Ser Thr
560 565 570
atg tca cca ctg ccc caa aat aca cca ttc gaa gct ttt aag act gta 2019
Met Ser Pro Leu Pro Gln Asn Thr Pro Phe Glu Ala Phe Lys Thr Val
575 580 585 590
gat tta cct tct act gtt act att aga tat act tct gct gct tca aca 2067
Asp Leu Pro Ser Thr Val Thr Ile Arg Tyr Thr Ser Ala Ala Ser Thr
595 600 605
act ttt caa ctt aat ttc cgt ttc act gtg cca gga agc gca aat gta 2115
Thr Phe Gln Leu Asn Phe Arg Phe Thr Val Pro Gly Ser Ala Asn Val
610 615 620
ttg att gac cga att gaa ttt gtt cca att gag ggt tcc ttg ttc gag 2163
Leu Ile Asp Arg Ile Glu Phe Val Pro Ile Glu Gly Ser Leu Phe Glu
625 630 635
tac gaa acc aaa cag cag cta gaa aaa gca agg aaa gcg gtg aac cat 2211
Tyr Glu Thr Lys Gln Gln Leu Glu Lys Ala Arg Lys Ala Val Asn His
640 645 650
ttg ttt aca gat gga tcg aaa aag gcg cta aaa gaa gac acg acc gat 2259
Leu Phe Thr Asp Gly Ser Lys Lys Ala Leu Lys Glu Asp Thr Thr Asp
655 660 665 670
tat gag att gat caa gcc gcc aac gtg gta gat tgt ata tcg gat gag 2307
Tyr Glu Ile Asp Gln Ala Ala Asn Val Val Asp Cys Ile Ser Asp Glu
675 680 685
tgt gga cat gag aaa atg atc ctg tta gat gaa gta aaa tat gca aaa 2355
Cys Gly His Glu Lys Met Ile Leu Leu Asp Glu Val Lys Tyr Ala Lys
690 695 700
caa ctc agc caa gcc cgc aat tta ctg ctc aat ggg aat ttc gat gat 2403
Gln Leu Ser Gln Ala Arg Asn Leu Leu Leu Asn Gly Asn Phe Asp Asp
705 710 715
cta tat cca gct ctg gag agg gag aat cca tgg aaa aca agt ccg aat 2451
Leu Tyr Pro Ala Leu Glu Arg Glu Asn Pro Trp Lys Thr Ser Pro Asn
720 725 730
gtt acg atc cgt caa gat aac ccg att ttt aaa ggc cat tat ctc agt 2499
Val Thr Ile Arg Gln Asp Asn Pro Ile Phe Lys Gly His Tyr Leu Ser
735 740 745 750
atg gcg ggt gcg aac gat atc gag gcc acc aat gat acg ttc ccc acg 2547
Met Ala Gly Ala Asn Asp Ile Glu Ala Thr Asn Asp Thr Phe Pro Thr
755 760 765
tat gtc tat caa aaa ata gat gaa gcc aaa tta aag cca tat aca cgg 2595
Tyr Val Tyr Gln Lys Ile Asp Glu Ala Lys Leu Lys Pro Tyr Thr Arg
770 775 780
tat aaa gtg cgc ggg ttt gtt ggc agc agc aaa gat ctg gag ctg ttg 2643
Tyr Lys Val Arg Gly Phe Val Gly Ser Ser Lys Asp Leu Glu Leu Leu
785 790 795
gtt aca cgc tat aat gaa gaa gtt gat gcg att tta gat gta ccg gat 2691
Val Thr Arg Tyr Asn Glu Glu Val Asp Ala Ile Leu Asp Val Pro Asp
800 805 810
aat atc ccg cat gcg ccg act cct gtc tgc ggt gaa ttt gat cga tgc 2739
Asn Ile Pro His Ala Pro Thr Pro Val Cys Gly Glu Phe Asp Arg Cys
815 820 825 830
aag ccc tat tcg tat cca cct tta ctt cca gaa tgt aac cct gag ttt 2787
Lys Pro Tyr Ser Tyr Pro Pro Leu Leu Pro Glu Cys Asn Pro Glu Phe
835 840 845
ata aat cag atg caa cca tcc tct tgc cac cac aat cag atg gtc gat 2835
Ile Asn Gln Met Gln Pro Ser Ser Cys His His Asn Gln Met Val Asp
850 855 860
tac aat aac atg aac acg agc acg agt act acc atg aat cct agc atg 2883
Tyr Asn Asn Met Asn Thr Ser Thr Ser Thr Thr Met Asn Pro Ser Met
865 870 875
aat cct ccc ctt acg cct gaa ata gca tcc agc caa agt gga ttc ggc 2931
Asn Pro Pro Leu Thr Pro Glu Ile Ala Ser Ser Gln Ser Gly Phe Gly
880 885 890
aga aaa cat cgc aaa tgt cat caa gcg cat caa ttt gag ttc cac att 2979
Arg Lys His Arg Lys Cys His Gln Ala His Gln Phe Glu Phe His Ile
895 900 905 910
gat acc ggg aca atc gat ttg gtg gaa gat ttg ggc att tgg gtg atc 3027
Asp Thr Gly Thr Ile Asp Leu Val Glu Asp Leu Gly Ile Trp Val Ile
915 920 925
ttc aaa atc tgt gcc aca gat gga tac gca agc tta gat gat ctg gaa 3075
Phe Lys Ile Cys Ala Thr Asp Gly Tyr Ala Ser Leu Asp Asp Leu Glu
930 935 940
gtg att gaa gaa gga gcg ctg ggg gtc gaa gcc tta gaa ctt gtc aag 3123
Val Ile Glu Glu Gly Ala Leu Gly Val Glu Ala Leu Glu Leu Val Lys
945 950 955
aaa aga gaa aag aaa tgg aga cat cag aag gag cag cac tgt tcg caa 3171
Lys Arg Glu Lys Lys Trp Arg His Gln Lys Glu Gln His Cys Ser Gln
960 965 970
acg aaa cac aaa tat gat gcg gcc aaa cat gcg gtg atg gcg tta ttt 3219
Thr Lys His Lys Tyr Asp Ala Ala Lys His Ala Val Met Ala Leu Phe
975 980 985 990
aca aac acg cgc tat gaa aaa ttg aag ttc gaa aca acc att tct gac 3267
Thr Asn Thr Arg Tyr Glu Lys Leu Lys Phe Glu Thr Thr Ile Ser Asp
995 1000 1005
att ttg tat gct gat cat ctc gtg cag tcg att cct tat gta tat aat 3315
Ile Leu Tyr Ala Asp His Leu Val Gln Ser Ile Pro Tyr Val Tyr Asn
1010 1015 1020
aaa tat gta ccg gaa gtt tca ggt atg aat tac gaa ctc tat aca gag 3363
Lys Tyr Val Pro Glu Val Ser Gly Met Asn Tyr Glu Leu Tyr Thr Glu
1025 1030 1035
cta aac act ctc gtt cag aat gcg ttc tac ctg tat gac cag cgg aat 3411
Leu Asn Thr Leu Val Gln Asn Ala Phe Tyr Leu Tyr Asp Gln Arg Asn
1040 1045 1050
ctg att aaa aat ggg cgc ttt agc aat ggg ctt atg tat tgg caa gct 3459
Leu Ile Lys Asn Gly Arg Phe Ser Asn Gly Leu Met Tyr Trp Gln Ala
1055 1060 1065 1070
acc ccg cat gca cga gtg gag caa gaa tat gat aga tca gtg ctg gtg 3507
Thr Pro His Ala Arg Val Glu Gln Glu Tyr Asp Arg Ser Val Leu Val
1075 1080 1085
ctg ccg aat tgg gat gcc aat gtg tcg caa cag ctg tgt atc gaa cac 3555
Leu Pro Asn Trp Asp Ala Asn Val Ser Gln Gln Leu Cys Ile Glu His
1090 1095 1100
aat cgc ggt tat gta ttg cgt gtc acg gcg aga aaa gaa gat ccg gga 3603
Asn Arg Gly Tyr Val Leu Arg Val Thr Ala Arg Lys Glu Asp Pro Gly
1105 1110 1115
gcc ggc aat gtt acc ttt agt gac tgt gca aat cat gtc gac aag ctg 3651
Ala Gly Asn Val Thr Phe Ser Asp Cys Ala Asn His Val Asp Lys Leu
1120 1125 1130
agc ttt act tct tgc gat ata gct aca aac gca gtg cca ggt gcc caa 3699
Ser Phe Thr Ser Cys Asp Ile Ala Thr Asn Ala Val Pro Gly Ala Gln
1135 1140 1145 1150
gcg aat gat cca gcc gcc gga gta gcc tat gga caa cag ggt tgt caa 3747
Ala Asn Asp Pro Ala Ala Gly Val Ala Tyr Gly Gln Gln Gly Cys Gln
1155 1160 1165
ata gat aga gtg ccg tac gga cca tct gga tat cga gca gac gga gta 3795
Ile Asp Arg Val Pro Tyr Gly Pro Ser Gly Tyr Arg Ala Asp Gly Val
1170 1175 1180
gcg tac gaa cag tct ggt cat cga aca gat gga gtg ccg tac aga caa 3843
Ala Tyr Glu Gln Ser Gly His Arg Thr Asp Gly Val Pro Tyr Arg Gln
1185 1190 1195
tct gga tat cga gca gac gga gta gcg cac gac caa cct gga tat cga 3891
Ser Gly Tyr Arg Ala Asp Gly Val Ala His Asp Gln Pro Gly Tyr Arg
1200 1205 1210
gca gac gga gta gcg tac gaa caa tct gga tat cga gca gat gga gta 3939
Ala Asp Gly Val Ala Tyr Glu Gln Ser Gly Tyr Arg Ala Asp Gly Val
1215 1220 1225 1230
gcg tac gaa cag tct ggt cat cga gca gat gga gtg ccg tac gga caa 3987
Ala Tyr Glu Gln Ser Gly His Arg Ala Asp Gly Val Pro Tyr Gly Gln
1235 1240 1245
tct gga tat gga aca gac gga gta acg tac gac caa tct gcc aaa caa 4035
Ser Gly Tyr Gly Thr Asp Gly Val Thr Tyr Asp Gln Ser Ala Lys Gln
1250 1255 1260
acc cgc aaa tac cat ggt tgc cat aca gac gga ctg cca cat cca gag 4083
Thr Arg Lys Tyr His Gly Cys His Thr Asp Gly Leu Pro His Pro Glu
1265 1270 1275
cat ggt tgt tgt tat cca gac aga gta agc gat ggc caa cag ctt gca 4131
His Gly Cys Cys Tyr Pro Asp Arg Val Ser Asp Gly Gln Gln Leu Ala
1280 1285 1290
tat gta aca aaa tcg att gat ctg ttc ccg gat aca gat aaa gtc cgg 4179
Tyr Val Thr Lys Ser Ile Asp Leu Phe Pro Asp Thr Asp Lys Val Arg
1295 1300 1305 1310
atc gac att gga gaa acc gaa ggg aac ttt aga gtg gaa agt gtg gaa 4227
Ile Asp Ile Gly Glu Thr Glu Gly Asn Phe Arg Val Glu Ser Val Glu
1315 1320 1325
ttg att tgt atg gaa aag taaatcatca caagtaaaag tatcgtttac 4275
Leu Ile Cys Met Glu Lys
1330
taaaaattta ttttccaagc aacaggggag aggggggctg tccagaaggt cagtgaaaac 4335
tggacgcccc gttttagtag aata 4359
<210> SEQ ID NO 18
<211> LENGTH: 1332
<212> TYPE: PRT
<213> ORGANISM: Bacillus popilliae
<400> SEQUENCE: 18
Met Gln Ile Ile Gln Pro Ser Ser Asn Ala Leu Leu Tyr Ser Pro Asn
1 5 10 15
Lys Tyr Pro Tyr Ala Thr Asp Pro Asn Val Ile Ala Glu Gly Arg Ser
20 25 30
Tyr Asn Asn Trp Leu Asp Thr Cys Val Gly Val Gly Asp Gly Thr Arg
35 40 45
Ser Pro Glu Ala Tyr Ala Ile Ala Glu Glu Ala Val Gly Leu Ser Ile
50 55 60
Asp Ile Leu Ala Glu Ile Ile Tyr Tyr Leu Gly Phe Pro Ile Ala Ser
65 70 75 80
Pro Leu Thr Arg Ala Leu Ser Ala Ile Ala Gly Gln Leu Phe Ser Ser
85 90 95
Gly Asp Thr Leu Met Gln His Ile Glu Gln Leu Ile Asn Gln Lys Ile
100 105 110
Ala Glu Tyr Ala Arg Asn Lys Ala Leu Ala Glu Phe Gln Gly Leu Gly
115 120 125
Arg Gln Tyr Gly Leu Tyr Leu Glu Ala Leu Glu Asp Trp Glu Gln Asn
130 135 140
Arg Leu Ser Gln Pro His Lys Glu Arg Val Arg Gln Thr Phe Arg Ile
145 150 155 160
Leu Asp Asn Ser Phe Thr Ser Ser Ile Pro Ser Phe Ala Val Arg Asn
165 170 175
Tyr Glu Val Pro Leu Leu Ser Val Tyr Ala Asp Ala Ala Asn Leu His
180 185 190
Leu Leu Ile Leu Arg Asp Ser Tyr Ile Tyr Gly Ala Phe Trp Gly Phe
195 200 205
Asp Glu Asp Glu Tyr Tyr Arg Asn Tyr Ala Arg Gln Ile Arg Leu Ser
210 215 220
Ala Glu Tyr Ala Asn His Cys Thr Thr Trp Tyr Gln Thr Gly Leu Arg
225 230 235 240
Arg Leu Gln Gly Thr Arg Ala Thr Asp Trp Ile Asn Tyr Asn Arg Phe
245 250 255
Arg Arg Glu Met Thr Leu Thr Val Leu Asp Ile Cys Ala Leu Phe Ser
260 265 270
Ser Tyr Asp Ile Pro Ser Tyr Pro Met Gly Thr Lys Ile Gln Leu Thr
275 280 285
Arg Glu Ile Tyr Thr Asp Pro Val Val His Ser Asp Trp Leu Gln Ser
290 295 300
Thr Ser Pro Gly Leu Ile Ser Phe Ser Ser Leu Glu Asn Leu Val Val
305 310 315 320
Arg Ala Pro His Leu Phe Thr Trp Leu Ser Arg Val Thr Ile Asp Thr
325 330 335
Gly Ile Leu Ser Thr Val Ile Gly Gly Gln Tyr Ser Asn Asn Asn Phe
340 345 350
Trp Arg Thr His Tyr Gln Thr Leu Arg Thr Thr Gly Gly Thr Ser Phe
355 360 365
Gln Ser Pro Thr Tyr Gly Ser Thr Ala Phe Pro Ile Gln Arg Thr Asn
370 375 380
Thr Leu Thr Phe Ser Gly Asp Val Tyr Thr Ile Glu Ser Ser Val Val
385 390 395 400
Thr Arg Ser Ser Leu Tyr Gly Ala Asn Ser Val Ala Phe Thr Gly Thr
405 410 415
Thr Gly Arg Ser Leu Tyr Glu Asn Pro Thr Val Tyr Pro Phe Ala Gln
420 425 430
Lys Leu Ile His Glu Leu Pro Gly Val Asp Ser Gly Arg Pro Asn Ala
435 440 445
Thr Asn Tyr Ser His Arg Leu Ser Tyr Ile Ser Gly Phe Ser Leu Gly
450 455 460
Tyr Ser Pro Ser Gly Thr Gly Leu Val Tyr Gly Trp Thr Ser Thr Thr
465 470 475 480
Ala Thr Arg Glu Asn Asn Ile Thr Leu Asp Asp Arg Ile Val Gln Leu
485 490 495
Pro Ala Val Lys Gly Ala Ser Leu Asn Asn Cys Gln Val Val Lys Gly
500 505 510
Thr Gly Phe Thr Gly Gly Asp Trp Leu Lys Pro Asn Asn Asn Gly Thr
515 520 525
Phe Ser Met Tyr Phe Ala Phe Arg Ser Ala Tyr Thr Tyr His Phe Arg
530 535 540
Ile Arg Tyr Ala Ser Ser Ala Ser Phe Ser Phe Val Ile Ser Glu Glu
545 550 555 560
Tyr Gly Arg Phe Pro Thr Thr Thr Val Pro Leu Leu Ser Thr Met Ser
565 570 575
Pro Leu Pro Gln Asn Thr Pro Phe Glu Ala Phe Lys Thr Val Asp Leu
580 585 590
Pro Ser Thr Val Thr Ile Arg Tyr Thr Ser Ala Ala Ser Thr Thr Phe
595 600 605
Gln Leu Asn Phe Arg Phe Thr Val Pro Gly Ser Ala Asn Val Leu Ile
610 615 620
Asp Arg Ile Glu Phe Val Pro Ile Glu Gly Ser Leu Phe Glu Tyr Glu
625 630 635 640
Thr Lys Gln Gln Leu Glu Lys Ala Arg Lys Ala Val Asn His Leu Phe
645 650 655
Thr Asp Gly Ser Lys Lys Ala Leu Lys Glu Asp Thr Thr Asp Tyr Glu
660 665 670
Ile Asp Gln Ala Ala Asn Val Val Asp Cys Ile Ser Asp Glu Cys Gly
675 680 685
His Glu Lys Met Ile Leu Leu Asp Glu Val Lys Tyr Ala Lys Gln Leu
690 695 700
Ser Gln Ala Arg Asn Leu Leu Leu Asn Gly Asn Phe Asp Asp Leu Tyr
705 710 715 720
Pro Ala Leu Glu Arg Glu Asn Pro Trp Lys Thr Ser Pro Asn Val Thr
725 730 735
Ile Arg Gln Asp Asn Pro Ile Phe Lys Gly His Tyr Leu Ser Met Ala
740 745 750
Gly Ala Asn Asp Ile Glu Ala Thr Asn Asp Thr Phe Pro Thr Tyr Val
755 760 765
Tyr Gln Lys Ile Asp Glu Ala Lys Leu Lys Pro Tyr Thr Arg Tyr Lys
770 775 780
Val Arg Gly Phe Val Gly Ser Ser Lys Asp Leu Glu Leu Leu Val Thr
785 790 795 800
Arg Tyr Asn Glu Glu Val Asp Ala Ile Leu Asp Val Pro Asp Asn Ile
805 810 815
Pro His Ala Pro Thr Pro Val Cys Gly Glu Phe Asp Arg Cys Lys Pro
820 825 830
Tyr Ser Tyr Pro Pro Leu Leu Pro Glu Cys Asn Pro Glu Phe Ile Asn
835 840 845
Gln Met Gln Pro Ser Ser Cys His His Asn Gln Met Val Asp Tyr Asn
850 855 860
Asn Met Asn Thr Ser Thr Ser Thr Thr Met Asn Pro Ser Met Asn Pro
865 870 875 880
Pro Leu Thr Pro Glu Ile Ala Ser Ser Gln Ser Gly Phe Gly Arg Lys
885 890 895
His Arg Lys Cys His Gln Ala His Gln Phe Glu Phe His Ile Asp Thr
900 905 910
Gly Thr Ile Asp Leu Val Glu Asp Leu Gly Ile Trp Val Ile Phe Lys
915 920 925
Ile Cys Ala Thr Asp Gly Tyr Ala Ser Leu Asp Asp Leu Glu Val Ile
930 935 940
Glu Glu Gly Ala Leu Gly Val Glu Ala Leu Glu Leu Val Lys Lys Arg
945 950 955 960
Glu Lys Lys Trp Arg His Gln Lys Glu Gln His Cys Ser Gln Thr Lys
965 970 975
His Lys Tyr Asp Ala Ala Lys His Ala Val Met Ala Leu Phe Thr Asn
980 985 990
Thr Arg Tyr Glu Lys Leu Lys Phe Glu Thr Thr Ile Ser Asp Ile Leu
995 1000 1005
Tyr Ala Asp His Leu Val Gln Ser Ile Pro Tyr Val Tyr Asn Lys Tyr
1010 1015 1020
Val Pro Glu Val Ser Gly Met Asn Tyr Glu Leu Tyr Thr Glu Leu Asn
1025 1030 1035 1040
Thr Leu Val Gln Asn Ala Phe Tyr Leu Tyr Asp Gln Arg Asn Leu Ile
1045 1050 1055
Lys Asn Gly Arg Phe Ser Asn Gly Leu Met Tyr Trp Gln Ala Thr Pro
1060 1065 1070
His Ala Arg Val Glu Gln Glu Tyr Asp Arg Ser Val Leu Val Leu Pro
1075 1080 1085
Asn Trp Asp Ala Asn Val Ser Gln Gln Leu Cys Ile Glu His Asn Arg
1090 1095 1100
Gly Tyr Val Leu Arg Val Thr Ala Arg Lys Glu Asp Pro Gly Ala Gly
1105 1110 1115 1120
Asn Val Thr Phe Ser Asp Cys Ala Asn His Val Asp Lys Leu Ser Phe
1125 1130 1135
Thr Ser Cys Asp Ile Ala Thr Asn Ala Val Pro Gly Ala Gln Ala Asn
1140 1145 1150
Asp Pro Ala Ala Gly Val Ala Tyr Gly Gln Gln Gly Cys Gln Ile Asp
1155 1160 1165
Arg Val Pro Tyr Gly Pro Ser Gly Tyr Arg Ala Asp Gly Val Ala Tyr
1170 1175 1180
Glu Gln Ser Gly His Arg Thr Asp Gly Val Pro Tyr Arg Gln Ser Gly
1185 1190 1195 1200
Tyr Arg Ala Asp Gly Val Ala His Asp Gln Pro Gly Tyr Arg Ala Asp
1205 1210 1215
Gly Val Ala Tyr Glu Gln Ser Gly Tyr Arg Ala Asp Gly Val Ala Tyr
1220 1225 1230
Glu Gln Ser Gly His Arg Ala Asp Gly Val Pro Tyr Gly Gln Ser Gly
1235 1240 1245
Tyr Gly Thr Asp Gly Val Thr Tyr Asp Gln Ser Ala Lys Gln Thr Arg
1250 1255 1260
Lys Tyr His Gly Cys His Thr Asp Gly Leu Pro His Pro Glu His Gly
1265 1270 1275 1280
Cys Cys Tyr Pro Asp Arg Val Ser Asp Gly Gln Gln Leu Ala Tyr Val
1285 1290 1295
Thr Lys Ser Ile Asp Leu Phe Pro Asp Thr Asp Lys Val Arg Ile Asp
1300 1305 1310
Ile Gly Glu Thr Glu Gly Asn Phe Arg Val Glu Ser Val Glu Leu Ile
1315 1320 1325
Cys Met Glu Lys
1330
<210> SEQ ID NO 19
<211> LENGTH: 4366
<212> TYPE: DNA
<213> ORGANISM: Bacillus popilliae
<220> FEATURE:
<221> NAME/KEY: CDS
<222> LOCATION: (224)..(4255)
<400> SEQUENCE: 19
gtagatcatg tactcaaatg cagtgtcgga agtttgccag atgttcatat cgattgtcaa 60
tatgtgacgg tatgtgattt acagatgagg cccgttcatg agggagcttg tcaatttgtg 120
aagattagcg gagaatttca attttattca ctttaacatt agctacaaat gttgtggcta 180
agcaccccat tatataaatg gatgacaacc aggaggggta act atg aat cag tat 235
Met Asn Gln Tyr
1
cat aac caa aac gat aac aaa agt tac aac caa agt gga aat gaa atg 283
His Asn Gln Asn Asp Asn Lys Ser Tyr Asn Gln Ser Gly Asn Glu Met
5 10 15 20
caa atc att caa cct tca agt aat tct tta ctt tac agt ccc aat aag 331
Gln Ile Ile Gln Pro Ser Ser Asn Ser Leu Leu Tyr Ser Pro Asn Lys
25 30 35
tat ccg tat gcc acg gat ccc aat gtc ata gca gag ggt aga agt tat 379
Tyr Pro Tyr Ala Thr Asp Pro Asn Val Ile Ala Glu Gly Arg Ser Tyr
40 45 50
aaa aat tgg ctt gat atg tgt gta ggt gaa ggc gac ggt aca cga agt 427
Lys Asn Trp Leu Asp Met Cys Val Gly Glu Gly Asp Gly Thr Arg Ser
55 60 65
ctc gag gct att gct gtt gct gtc gga gtt cga ata agc cac aca att 475
Leu Glu Ala Ile Ala Val Ala Val Gly Val Arg Ile Ser His Thr Ile
70 75 80
ttc cgc ctt tta ggt gtt cca tat tca gct caa ggc gag caa tta ttt 523
Phe Arg Leu Leu Gly Val Pro Tyr Ser Ala Gln Gly Glu Gln Leu Phe
85 90 95 100
agc ttc cta ttg gat acg tta tgg ctt gaa ggc aat act caa tgg gaa 571
Ser Phe Leu Leu Asp Thr Leu Trp Leu Glu Gly Asn Thr Gln Trp Glu
105 110 115
gag ttg atg aga cat gca gaa gaa ctc ata aat gaa cag gta ccg gat 619
Glu Leu Met Arg His Ala Glu Glu Leu Ile Asn Glu Gln Val Pro Asp
120 125 130
tat gta aga acc aag gca ctt gca gaa tta acg gat tta ggt aac aac 667
Tyr Val Arg Thr Lys Ala Leu Ala Glu Leu Thr Asp Leu Gly Asn Asn
135 140 145
tta aat tta tat ata gca gct ttt gaa gat tgg aaa cga aat ccg agc 715
Leu Asn Leu Tyr Ile Ala Ala Phe Glu Asp Trp Lys Arg Asn Pro Ser
150 155 160
agt caa gaa gtt aga acc cgg gta ata gat aga ttc aat ata ctc gac 763
Ser Gln Glu Val Arg Thr Arg Val Ile Asp Arg Phe Asn Ile Leu Asp
165 170 175 180
ggt tta ttt gaa gcc tat ctg cct tca ttt gca gta cct ggt tat gaa 811
Gly Leu Phe Glu Ala Tyr Leu Pro Ser Phe Ala Val Pro Gly Tyr Glu
185 190 195
gta cca cta tta tcc gtg tat gca aat gtt gta aat atc cac tta ttg 859
Val Pro Leu Leu Ser Val Tyr Ala Asn Val Val Asn Ile His Leu Leu
200 205 210
gta ctg agg gac agc tcg att tat ggt ctg gat tgg gga tta agt tca 907
Val Leu Arg Asp Ser Ser Ile Tyr Gly Leu Asp Trp Gly Leu Ser Ser
215 220 225
act agt gtt gac aat aat tac aat cgc caa caa agg aac tcc gca acg 955
Thr Ser Val Asp Asn Asn Tyr Asn Arg Gln Gln Arg Asn Ser Ala Thr
230 235 240
tat gca aat cat tgt aca act tgg tat cag acg ggt tta caa aga ttg 1003
Tyr Ala Asn His Cys Thr Thr Trp Tyr Gln Thr Gly Leu Gln Arg Leu
245 250 255 260
caa ggc agc gat gct agc agt tgg gtc aat tat aat cga ttt aga aga 1051
Gln Gly Ser Asp Ala Ser Ser Trp Val Asn Tyr Asn Arg Phe Arg Arg
265 270 275
gaa ata acg tta ata gta ttg gat att tgt gca ttg ttt tca aat tat 1099
Glu Ile Thr Leu Ile Val Leu Asp Ile Cys Ala Leu Phe Ser Asn Tyr
280 285 290
gat gtt cgt agt tat cca ata cag tta cgg gga gag ctt acg aga gga 1147
Asp Val Arg Ser Tyr Pro Ile Gln Leu Arg Gly Glu Leu Thr Arg Gly
295 300 305
att tat acg gat cca gca gta tat agc ggt aca ggt tcc tat tcc tgg 1195
Ile Tyr Thr Asp Pro Ala Val Tyr Ser Gly Thr Gly Ser Tyr Ser Trp
310 315 320
ttg agt caa gca cca tca ttt gca gaa ata gaa aat atc gca att agg 1243
Leu Ser Gln Ala Pro Ser Phe Ala Glu Ile Glu Asn Ile Ala Ile Arg
325 330 335 340
gaa cca agc aat ttt act tgg gca tct tat gcg aga gta aca aca ggt 1291
Glu Pro Ser Asn Phe Thr Trp Ala Ser Tyr Ala Arg Val Thr Thr Gly
345 350 355
aca ctg gaa tat ctc agc tct aag aat gat ttt tgg aaa tca cac tat 1339
Thr Leu Glu Tyr Leu Ser Ser Lys Asn Asp Phe Trp Lys Ser His Tyr
360 365 370
atg aac tat act gaa acc aat tcg ggt ata ttg att caa gga cct acc 1387
Met Asn Tyr Thr Glu Thr Asn Ser Gly Ile Leu Ile Gln Gly Pro Thr
375 380 385
tat gga atg acg acg ggt aca aat att cgt ata gag tcc gta tca atg 1435
Tyr Gly Met Thr Thr Gly Thr Asn Ile Arg Ile Glu Ser Val Ser Met
390 395 400
caa gaa att tat tcc gtt aga tta gaa gct gtt gct cat gct gga gct 1483
Gln Glu Ile Tyr Ser Val Arg Leu Glu Ala Val Ala His Ala Gly Ala
405 410 415 420
ggg ggt cct ttt ttg gga atc tct acg tct gaa ttt ttc tgg agt ttg 1531
Gly Gly Pro Phe Leu Gly Ile Ser Thr Ser Glu Phe Phe Trp Ser Leu
425 430 435
ggt gtt aga agg tat cag aac tca cgt agt cct caa ttt gcg tct caa 1579
Gly Val Arg Arg Tyr Gln Asn Ser Arg Ser Pro Gln Phe Ala Ser Gln
440 445 450
ata ata act agg caa tta cct gga gta aac tca gcg gtt cca tct gcc 1627
Ile Ile Thr Arg Gln Leu Pro Gly Val Asn Ser Ala Val Pro Ser Ala
455 460 465
ctc gac cat agt cat gaa cta tcg tat atc aca gcg ttt cca gtt aga 1675
Leu Asp His Ser His Glu Leu Ser Tyr Ile Thr Ala Phe Pro Val Arg
470 475 480
tcg gtg gga acg att ctc gtt cat gaa tgg aca tct aca aca gtt agt 1723
Ser Val Gly Thr Ile Leu Val His Glu Trp Thr Ser Thr Thr Val Ser
485 490 495 500
cgt aac aat aga att gag cca gat aaa ata aca caa atc ccg gct gtt 1771
Arg Asn Asn Arg Ile Glu Pro Asp Lys Ile Thr Gln Ile Pro Ala Val
505 510 515
aag tca cac aca ctc tcc aat tgt caa gta gtt agt ggg act ggg ttt 1819
Lys Ser His Thr Leu Ser Asn Cys Gln Val Val Ser Gly Thr Gly Phe
520 525 530
acg gga gga aac tgg ttg aga cct tct gat aat ggt tca ttt aga cta 1867
Thr Gly Gly Asn Trp Leu Arg Pro Ser Asp Asn Gly Ser Phe Arg Leu
535 540 545
acg att act tca ttc tca agc caa tct tac cgc att cgc att cat tat 1915
Thr Ile Thr Ser Phe Ser Ser Gln Ser Tyr Arg Ile Arg Ile His Tyr
550 555 560
gct tcc gca aca ttt ttt tat ttg gat att cgt acg ggt gat act tct 1963
Ala Ser Ala Thr Phe Phe Tyr Leu Asp Ile Arg Thr Gly Asp Thr Ser
565 570 575 580
aac aca ttt gcg gtt acc cca aca aca tta tca tca gga tcc caa act 2011
Asn Thr Phe Ala Val Thr Pro Thr Thr Leu Ser Ser Gly Ser Gln Thr
585 590 595
gta ccc tac gaa tct ttt ggg ttt ata aat ata cct tat act ttt aca 2059
Val Pro Tyr Glu Ser Phe Gly Phe Ile Asn Ile Pro Tyr Thr Phe Thr
600 605 610
aca gca cct act gaa agt aga tat act ttt gat ttc atg ttc tac tca 2107
Thr Ala Pro Thr Glu Ser Arg Tyr Thr Phe Asp Phe Met Phe Tyr Ser
615 620 625
ata gga agc gca aat gta tta att gac cga att gaa att gtt cca atc 2155
Ile Gly Ser Ala Asn Val Leu Ile Asp Arg Ile Glu Ile Val Pro Ile
630 635 640
gga gtt cct ttg ttc gag tac gaa acc aaa cag cag cta gaa aaa gca 2203
Gly Val Pro Leu Phe Glu Tyr Glu Thr Lys Gln Gln Leu Glu Lys Ala
645 650 655 660
agg aaa gcg gtg aac cat ttg ttt aca gat gga tcg aaa aag gcg cta 2251
Arg Lys Ala Val Asn His Leu Phe Thr Asp Gly Ser Lys Lys Ala Leu
665 670 675
aaa gaa gac acg acc gat tat gag att gat caa gcc gcc aac gtg gta 2299
Lys Glu Asp Thr Thr Asp Tyr Glu Ile Asp Gln Ala Ala Asn Val Val
680 685 690
gat tgt ata tcg gat gag tgt gga cat gat aaa atg atc ctg tta gat 2347
Asp Cys Ile Ser Asp Glu Cys Gly His Asp Lys Met Ile Leu Leu Asp
695 700 705
gaa gta aaa tat gca aaa caa ctc agc caa gcc cgc aat tta ctg ctc 2395
Glu Val Lys Tyr Ala Lys Gln Leu Ser Gln Ala Arg Asn Leu Leu Leu
710 715 720
aat ggg aat ttc gat gat cta tat tca gct ctg gag aag gag aat cca 2443
Asn Gly Asn Phe Asp Asp Leu Tyr Ser Ala Leu Glu Lys Glu Asn Pro
725 730 735 740
tgg aaa aca agt ccg aat gtt acg atc cga caa gat aac ccg att ttt 2491
Trp Lys Thr Ser Pro Asn Val Thr Ile Arg Gln Asp Asn Pro Ile Phe
745 750 755
aaa ggc cat tat ctc agt atg gcg ggt gcg aac gat atc gag gcc acc 2539
Lys Gly His Tyr Leu Ser Met Ala Gly Ala Asn Asp Ile Glu Ala Thr
760 765 770
aat gat acc ttc ccc acg tat gtc tat caa aaa ata gac gaa gcc aaa 2587
Asn Asp Thr Phe Pro Thr Tyr Val Tyr Gln Lys Ile Asp Glu Ala Lys
775 780 785
tta aag ccg tat aca cgt tat aaa gtg cgc ggg ttt gtt ggc agc agc 2635
Leu Lys Pro Tyr Thr Arg Tyr Lys Val Arg Gly Phe Val Gly Ser Ser
790 795 800
aaa gct cta gag ctg ttg gtt aca cgc tat aat gaa gaa gtt gat gcg 2683
Lys Ala Leu Glu Leu Leu Val Thr Arg Tyr Asn Glu Glu Val Asp Ala
805 810 815 820
att tta gat gta ccg gat aat atc ccg cat gcg ccg act cct gtc tgc 2731
Ile Leu Asp Val Pro Asp Asn Ile Pro His Ala Pro Thr Pro Val Cys
825 830 835
ggt gaa ttt gat cga tgc aag ccc tat tcg tat cca cct tta ctt cca 2779
Gly Glu Phe Asp Arg Cys Lys Pro Tyr Ser Tyr Pro Pro Leu Leu Pro
840 845 850
gaa tgt aac cct gag ttt ata aat cag atg caa cca tcc tct tgc cac 2827
Glu Cys Asn Pro Glu Phe Ile Asn Gln Met Gln Pro Ser Ser Cys His
855 860 865
cac aat cag atg gtc gat tac aat aac atg aac acg agc acg agt act 2875
His Asn Gln Met Val Asp Tyr Asn Asn Met Asn Thr Ser Thr Ser Thr
870 875 880
acc atg aat cct agc atg aat cct ccc ctt acg cct gaa ata gca tcc 2923
Thr Met Asn Pro Ser Met Asn Pro Pro Leu Thr Pro Glu Ile Ala Ser
885 890 895 900
agc caa agt gga ttc ggc aga aaa cat cgc aaa tgt cat caa gcg cat 2971
Ser Gln Ser Gly Phe Gly Arg Lys His Arg Lys Cys His Gln Ala His
905 910 915
caa ttt gag ttc cac att gat acc ggg aca atc gat ttg gtc gaa gat 3019
Gln Phe Glu Phe His Ile Asp Thr Gly Thr Ile Asp Leu Val Glu Asp
920 925 930
ttg ggc att tgg gtg atc ttc aaa atc tgt gcc aca gat gga tac gca 3067
Leu Gly Ile Trp Val Ile Phe Lys Ile Cys Ala Thr Asp Gly Tyr Ala
935 940 945
agc tta gat gat ctg gaa gtg att gaa gaa gga gcg ctg ggt gtc gaa 3115
Ser Leu Asp Asp Leu Glu Val Ile Glu Glu Gly Ala Leu Gly Val Glu
950 955 960
gca tta gaa ctt gtc aaa aaa aga gaa aag aaa tgg aga cat cag aag 3163
Ala Leu Glu Leu Val Lys Lys Arg Glu Lys Lys Trp Arg His Gln Lys
965 970 975 980
gag cag cac tgt tcg caa acg aaa cac aaa tat gat gcg gcc aaa cat 3211
Glu Gln His Cys Ser Gln Thr Lys His Lys Tyr Asp Ala Ala Lys His
985 990 995
gcg gtg atg gcg tta ttt aca aac aag cgc tat gaa aaa ttg aag ttc 3259
Ala Val Met Ala Leu Phe Thr Asn Lys Arg Tyr Glu Lys Leu Lys Phe
1000 1005 1010
gaa aca acc att tct gac att ttg tat gct gat cat ctc gtg cag tcg 3307
Glu Thr Thr Ile Ser Asp Ile Leu Tyr Ala Asp His Leu Val Gln Ser
1015 1020 1025
att cct tat gta tat aat aaa tat gta ccg gaa gtt cca ggt atg aat 3355
Ile Pro Tyr Val Tyr Asn Lys Tyr Val Pro Glu Val Pro Gly Met Asn
1030 1035 1040
tac gaa ctc tat tca gag cta aac aca ctg gtt cag aat gcg ttc tac 3403
Tyr Glu Leu Tyr Ser Glu Leu Asn Thr Leu Val Gln Asn Ala Phe Tyr
1045 1050 1055 1060
ctg tat gac cag cgg aat ctg att aaa aat ggg cgc ttt agc aat ggg 3451
Leu Tyr Asp Gln Arg Asn Leu Ile Lys Asn Gly Arg Phe Ser Asn Gly
1065 1070 1075
ctt atg cat tgg caa gct act cct cat gca aga gta gag caa gaa tat 3499
Leu Met His Trp Gln Ala Thr Pro His Ala Arg Val Glu Gln Glu Tyr
1080 1085 1090
gag aaa tcg gtg ctc gtg ctg cca aat tgg gat gcc aat gtg tcg caa 3547
Glu Lys Ser Val Leu Val Leu Pro Asn Trp Asp Ala Asn Val Ser Gln
1095 1100 1105
gat ctt tgt atc gaa cac aat cgc ggt tat gta ttg cgt gtc acg gcg 3595
Asp Leu Cys Ile Glu His Asn Arg Gly Tyr Val Leu Arg Val Thr Ala
1110 1115 1120
aga aaa gaa gat ccg gga gct ggc aat gtt acc ttt agt gac tgt gaa 3643
Arg Lys Glu Asp Pro Gly Ala Gly Asn Val Thr Phe Ser Asp Cys Glu
1125 1130 1135 1140
aat cat gtc gac aag ctg agc ttt act tct tgc gat ata gct aca aac 3691
Asn His Val Asp Lys Leu Ser Phe Thr Ser Cys Asp Ile Ala Thr Asn
1145 1150 1155
gca gtg cca ggt gcc caa gcg aat gat cca gcc gcc gga gta gcc tat 3739
Ala Val Pro Gly Ala Gln Ala Asn Asp Pro Ala Ala Gly Val Ala Tyr
1160 1165 1170
gga caa cag ggt tgt caa ata gat aga gtg ccg tac ggg caa tct gga 3787
Gly Gln Gln Gly Cys Gln Ile Asp Arg Val Pro Tyr Gly Gln Ser Gly
1175 1180 1185
tat cga gca gac gga gta gcg tac gaa cag tct ggt cat cga aca gat 3835
Tyr Arg Ala Asp Gly Val Ala Tyr Glu Gln Ser Gly His Arg Thr Asp
1190 1195 1200
gga gtg ccg tac aga caa tct gga tat gga aca gac gga gta acg tac 3883
Gly Val Pro Tyr Arg Gln Ser Gly Tyr Gly Thr Asp Gly Val Thr Tyr
1205 1210 1215 1220
gaa caa tct ggt cat cga gca gat gga gtg ccg tac gga caa tct gga 3931
Glu Gln Ser Gly His Arg Ala Asp Gly Val Pro Tyr Gly Gln Ser Gly
1225 1230 1235
tat cga gca gat gga gta gcg tac gaa cag tct ggt cat cga gca gat 3979
Tyr Arg Ala Asp Gly Val Ala Tyr Glu Gln Ser Gly His Arg Ala Asp
1240 1245 1250
gga gtg ccg tac gga caa tct gga tat gga aca gac gga gta acg tac 4027
Gly Val Pro Tyr Gly Gln Ser Gly Tyr Gly Thr Asp Gly Val Thr Tyr
1255 1260 1265
gac caa tct gcc aat caa acc cgc aaa tat cat ggt tgc cat aca gac 4075
Asp Gln Ser Ala Asn Gln Thr Arg Lys Tyr His Gly Cys His Thr Asp
1270 1275 1280
gga ctg cca cat cca gag cat ggt tgt tgt tat cca gac aga gta agc 4123
Gly Leu Pro His Pro Glu His Gly Cys Cys Tyr Pro Asp Arg Val Ser
1285 1290 1295 1300
gat ggc caa cag ctt gca tat gta aca aaa tcg att gat ctg ttc ccg 4171
Asp Gly Gln Gln Leu Ala Tyr Val Thr Lys Ser Ile Asp Leu Phe Pro
1305 1310 1315
gat aca gat aaa gtc cgg atc gac att gga gaa acc gaa ggg aac ttt 4219
Asp Thr Asp Lys Val Arg Ile Asp Ile Gly Glu Thr Glu Gly Asn Phe
1320 1325 1330
aga gtg gaa agt gtg gaa ttg att tgt atg gaa aag taaatcatca 4265
Arg Val Glu Ser Val Glu Leu Ile Cys Met Glu Lys
1335 1340
caagtaaaag tatcgtttac taaaaattta ttttccaagc aacaggggag aagatgattt 4325
gggtgtaata ctcaaatcat cttttcttat aagccacttt a 4366
<210> SEQ ID NO 20
<211> LENGTH: 1344
<212> TYPE: PRT
<213> ORGANISM: Bacillus popilliae
<400> SEQUENCE: 20
Met Asn Gln Tyr His Asn Gln Asn Asp Asn Lys Ser Tyr Asn Gln Ser
1 5 10 15
Gly Asn Glu Met Gln Ile Ile Gln Pro Ser Ser Asn Ser Leu Leu Tyr
20 25 30
Ser Pro Asn Lys Tyr Pro Tyr Ala Thr Asp Pro Asn Val Ile Ala Glu
35 40 45
Gly Arg Ser Tyr Lys Asn Trp Leu Asp Met Cys Val Gly Glu Gly Asp
50 55 60
Gly Thr Arg Ser Leu Glu Ala Ile Ala Val Ala Val Gly Val Arg Ile
65 70 75 80
Ser His Thr Ile Phe Arg Leu Leu Gly Val Pro Tyr Ser Ala Gln Gly
85 90 95
Glu Gln Leu Phe Ser Phe Leu Leu Asp Thr Leu Trp Leu Glu Gly Asn
100 105 110
Thr Gln Trp Glu Glu Leu Met Arg His Ala Glu Glu Leu Ile Asn Glu
115 120 125
Gln Val Pro Asp Tyr Val Arg Thr Lys Ala Leu Ala Glu Leu Thr Asp
130 135 140
Leu Gly Asn Asn Leu Asn Leu Tyr Ile Ala Ala Phe Glu Asp Trp Lys
145 150 155 160
Arg Asn Pro Ser Ser Gln Glu Val Arg Thr Arg Val Ile Asp Arg Phe
165 170 175
Asn Ile Leu Asp Gly Leu Phe Glu Ala Tyr Leu Pro Ser Phe Ala Val
180 185 190
Pro Gly Tyr Glu Val Pro Leu Leu Ser Val Tyr Ala Asn Val Val Asn
195 200 205
Ile His Leu Leu Val Leu Arg Asp Ser Ser Ile Tyr Gly Leu Asp Trp
210 215 220
Gly Leu Ser Ser Thr Ser Val Asp Asn Asn Tyr Asn Arg Gln Gln Arg
225 230 235 240
Asn Ser Ala Thr Tyr Ala Asn His Cys Thr Thr Trp Tyr Gln Thr Gly
245 250 255
Leu Gln Arg Leu Gln Gly Ser Asp Ala Ser Ser Trp Val Asn Tyr Asn
260 265 270
Arg Phe Arg Arg Glu Ile Thr Leu Ile Val Leu Asp Ile Cys Ala Leu
275 280 285
Phe Ser Asn Tyr Asp Val Arg Ser Tyr Pro Ile Gln Leu Arg Gly Glu
290 295 300
Leu Thr Arg Gly Ile Tyr Thr Asp Pro Ala Val Tyr Ser Gly Thr Gly
305 310 315 320
Ser Tyr Ser Trp Leu Ser Gln Ala Pro Ser Phe Ala Glu Ile Glu Asn
325 330 335
Ile Ala Ile Arg Glu Pro Ser Asn Phe Thr Trp Ala Ser Tyr Ala Arg
340 345 350
Val Thr Thr Gly Thr Leu Glu Tyr Leu Ser Ser Lys Asn Asp Phe Trp
355 360 365
Lys Ser His Tyr Met Asn Tyr Thr Glu Thr Asn Ser Gly Ile Leu Ile
370 375 380
Gln Gly Pro Thr Tyr Gly Met Thr Thr Gly Thr Asn Ile Arg Ile Glu
385 390 395 400
Ser Val Ser Met Gln Glu Ile Tyr Ser Val Arg Leu Glu Ala Val Ala
405 410 415
His Ala Gly Ala Gly Gly Pro Phe Leu Gly Ile Ser Thr Ser Glu Phe
420 425 430
Phe Trp Ser Leu Gly Val Arg Arg Tyr Gln Asn Ser Arg Ser Pro Gln
435 440 445
Phe Ala Ser Gln Ile Ile Thr Arg Gln Leu Pro Gly Val Asn Ser Ala
450 455 460
Val Pro Ser Ala Leu Asp His Ser His Glu Leu Ser Tyr Ile Thr Ala
465 470 475 480
Phe Pro Val Arg Ser Val Gly Thr Ile Leu Val His Glu Trp Thr Ser
485 490 495
Thr Thr Val Ser Arg Asn Asn Arg Ile Glu Pro Asp Lys Ile Thr Gln
500 505 510
Ile Pro Ala Val Lys Ser His Thr Leu Ser Asn Cys Gln Val Val Ser
515 520 525
Gly Thr Gly Phe Thr Gly Gly Asn Trp Leu Arg Pro Ser Asp Asn Gly
530 535 540
Ser Phe Arg Leu Thr Ile Thr Ser Phe Ser Ser Gln Ser Tyr Arg Ile
545 550 555 560
Arg Ile His Tyr Ala Ser Ala Thr Phe Phe Tyr Leu Asp Ile Arg Thr
565 570 575
Gly Asp Thr Ser Asn Thr Phe Ala Val Thr Pro Thr Thr Leu Ser Ser
580 585 590
Gly Ser Gln Thr Val Pro Tyr Glu Ser Phe Gly Phe Ile Asn Ile Pro
595 600 605
Tyr Thr Phe Thr Thr Ala Pro Thr Glu Ser Arg Tyr Thr Phe Asp Phe
610 615 620
Met Phe Tyr Ser Ile Gly Ser Ala Asn Val Leu Ile Asp Arg Ile Glu
625 630 635 640
Ile Val Pro Ile Gly Val Pro Leu Phe Glu Tyr Glu Thr Lys Gln Gln
645 650 655
Leu Glu Lys Ala Arg Lys Ala Val Asn His Leu Phe Thr Asp Gly Ser
660 665 670
Lys Lys Ala Leu Lys Glu Asp Thr Thr Asp Tyr Glu Ile Asp Gln Ala
675 680 685
Ala Asn Val Val Asp Cys Ile Ser Asp Glu Cys Gly His Asp Lys Met
690 695 700
Ile Leu Leu Asp Glu Val Lys Tyr Ala Lys Gln Leu Ser Gln Ala Arg
705 710 715 720
Asn Leu Leu Leu Asn Gly Asn Phe Asp Asp Leu Tyr Ser Ala Leu Glu
725 730 735
Lys Glu Asn Pro Trp Lys Thr Ser Pro Asn Val Thr Ile Arg Gln Asp
740 745 750
Asn Pro Ile Phe Lys Gly His Tyr Leu Ser Met Ala Gly Ala Asn Asp
755 760 765
Ile Glu Ala Thr Asn Asp Thr Phe Pro Thr Tyr Val Tyr Gln Lys Ile
770 775 780
Asp Glu Ala Lys Leu Lys Pro Tyr Thr Arg Tyr Lys Val Arg Gly Phe
785 790 795 800
Val Gly Ser Ser Lys Ala Leu Glu Leu Leu Val Thr Arg Tyr Asn Glu
805 810 815
Glu Val Asp Ala Ile Leu Asp Val Pro Asp Asn Ile Pro His Ala Pro
820 825 830
Thr Pro Val Cys Gly Glu Phe Asp Arg Cys Lys Pro Tyr Ser Tyr Pro
835 840 845
Pro Leu Leu Pro Glu Cys Asn Pro Glu Phe Ile Asn Gln Met Gln Pro
850 855 860
Ser Ser Cys His His Asn Gln Met Val Asp Tyr Asn Asn Met Asn Thr
865 870 875 880
Ser Thr Ser Thr Thr Met Asn Pro Ser Met Asn Pro Pro Leu Thr Pro
885 890 895
Glu Ile Ala Ser Ser Gln Ser Gly Phe Gly Arg Lys His Arg Lys Cys
900 905 910
His Gln Ala His Gln Phe Glu Phe His Ile Asp Thr Gly Thr Ile Asp
915 920 925
Leu Val Glu Asp Leu Gly Ile Trp Val Ile Phe Lys Ile Cys Ala Thr
930 935 940
Asp Gly Tyr Ala Ser Leu Asp Asp Leu Glu Val Ile Glu Glu Gly Ala
945 950 955 960
Leu Gly Val Glu Ala Leu Glu Leu Val Lys Lys Arg Glu Lys Lys Trp
965 970 975
Arg His Gln Lys Glu Gln His Cys Ser Gln Thr Lys His Lys Tyr Asp
980 985 990
Ala Ala Lys His Ala Val Met Ala Leu Phe Thr Asn Lys Arg Tyr Glu
995 1000 1005
Lys Leu Lys Phe Glu Thr Thr Ile Ser Asp Ile Leu Tyr Ala Asp His
1010 1015 1020
Leu Val Gln Ser Ile Pro Tyr Val Tyr Asn Lys Tyr Val Pro Glu Val
1025 1030 1035 1040
Pro Gly Met Asn Tyr Glu Leu Tyr Ser Glu Leu Asn Thr Leu Val Gln
1045 1050 1055
Asn Ala Phe Tyr Leu Tyr Asp Gln Arg Asn Leu Ile Lys Asn Gly Arg
1060 1065 1070
Phe Ser Asn Gly Leu Met His Trp Gln Ala Thr Pro His Ala Arg Val
1075 1080 1085
Glu Gln Glu Tyr Glu Lys Ser Val Leu Val Leu Pro Asn Trp Asp Ala
1090 1095 1100
Asn Val Ser Gln Asp Leu Cys Ile Glu His Asn Arg Gly Tyr Val Leu
1105 1110 1115 1120
Arg Val Thr Ala Arg Lys Glu Asp Pro Gly Ala Gly Asn Val Thr Phe
1125 1130 1135
Ser Asp Cys Glu Asn His Val Asp Lys Leu Ser Phe Thr Ser Cys Asp
1140 1145 1150
Ile Ala Thr Asn Ala Val Pro Gly Ala Gln Ala Asn Asp Pro Ala Ala
1155 1160 1165
Gly Val Ala Tyr Gly Gln Gln Gly Cys Gln Ile Asp Arg Val Pro Tyr
1170 1175 1180
Gly Gln Ser Gly Tyr Arg Ala Asp Gly Val Ala Tyr Glu Gln Ser Gly
1185 1190 1195 1200
His Arg Thr Asp Gly Val Pro Tyr Arg Gln Ser Gly Tyr Gly Thr Asp
1205 1210 1215
Gly Val Thr Tyr Glu Gln Ser Gly His Arg Ala Asp Gly Val Pro Tyr
1220 1225 1230
Gly Gln Ser Gly Tyr Arg Ala Asp Gly Val Ala Tyr Glu Gln Ser Gly
1235 1240 1245
His Arg Ala Asp Gly Val Pro Tyr Gly Gln Ser Gly Tyr Gly Thr Asp
1250 1255 1260
Gly Val Thr Tyr Asp Gln Ser Ala Asn Gln Thr Arg Lys Tyr His Gly
1265 1270 1275 1280
Cys His Thr Asp Gly Leu Pro His Pro Glu His Gly Cys Cys Tyr Pro
1285 1290 1295
Asp Arg Val Ser Asp Gly Gln Gln Leu Ala Tyr Val Thr Lys Ser Ile
1300 1305 1310
Asp Leu Phe Pro Asp Thr Asp Lys Val Arg Ile Asp Ile Gly Glu Thr
1315 1320 1325
Glu Gly Asn Phe Arg Val Glu Ser Val Glu Leu Ile Cys Met Glu Lys
1330 1335 1340
<210> SEQ ID NO 21
<211> LENGTH: 34
<212> TYPE: DNA
<213> ORGANISM: Artificial Sequence
<220> FEATURE:
<223> OTHER INFORMATION: synthetic oligonucleotide
<400> SEQUENCE: 21
ctggaattca tgaatcagta tcataaccaa aacg 34
<210> SEQ ID NO 22
<211> LENGTH: 34
<212> TYPE: DNA
<213> ORGANISM: Artificial Sequence
<220> FEATURE:
<223> OTHER INFORMATION: synthetic oligonucleotide
<400> SEQUENCE: 22
gagctgcagt tacttttcca tacaaatcaa ttcc 34
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