Patent application title: TRANSGENIC PLANTS WITH ENHANCED TRAITS
Inventors:
IPC8 Class: AC12N1582FI
USPC Class:
1 1
Class name:
Publication date: 2022-04-07
Patent application number: 20220106606
Abstract:
This disclosure provides recombinant DNA constructs and transgenic plants
having enhanced traits such as increased yield, increased nitrogen use
efficiency and enhanced drought tolerance; propagules, progeny and field
crops of such transgenic plants; and methods of making and using such
transgenic plants. This disclosure also provides methods of producing
seed from such transgenic plants, growing such seed and selecting progeny
plants with enhanced traits. Also disclosed are transgenic plants with
altered phenotypes which are useful for screening and selecting
transgenic events for the desired enhanced trait.Claims:
1. A recombinant DNA construct comprising a heterologous promoter
functional in a plant cell and operably linked to: a) a polynucleotide
that comprises a nucleotide sequence with at least 90%, at least 91%, at
least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99% identity, or 100% identity to a
sequence selected from the group consisting of SEQ ID NOs: 1-27 and 55;
b) a DNA encoding RNA for suppressing the expression of a target mRNA
transcribed from a polynucleotide having a nucleic acid sequence with at
least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%
identity, or 100% identity to a sequence selected from the group
consisting of SEQ ID NOs: 6 and 55-60; c) a polynucleotide that encodes a
polypeptide having an amino acid sequence with at least 90%, at least
91%, at least 92%, at least 93%, at least 94%, at least 95%, at least
96%, at least 97%, at least 98%, at least 99% identity, or 100% identity
to a sequence selected from the group consisting of SEQ ID NOs: 28-54,
61, 79-95 and 96; or d) a DNA encoding RNA for suppressing the expression
of a target protein having an amino acid sequence with at least 90%, at
least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at
least 96%, at least 97%, at least 98%, at least 99% identity, or 100%
identity to a sequence selected from the group consisting of SEQ ID NOs:
33, 61-66, 82, and 96.
2. The recombinant DNA construct of claim 1, wherein said RNA is a double-stranded RNA, an antisense RNA, a miRNA precursor, or a ta-siRNA.
3. The recombinant DNA construct of claim 1, wherein said RNA is a miRNA precursor that produces a mature miRNA, and wherein said mature miRNA has a nucleic acid sequence with at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity or complementarity to a fragment of at least 19, 20, 21, 22, 23, 24, 25, 26 or 27 consecutive nucleotides of a sequence selected from the group consisting of SEQ ID NOs: 6 and 55-60.
4. The recombinant DNA construct of claim 1, wherein said RNA is a miRNA precursor that produces a mature miRNA having a nucleic acid sequence with 100% identity or 100% complementarity to a fragment of 21 consecutive nucleotides of a sequence selected from the group consisting of SEQ ID NOs: 6 and 55-60.
5. The recombinant DNA construct of claim 1, wherein said DNA comprises a sequence selected from the group consisting of SEQ ID NOs: 67-72.
6. A plant comprising a recombinant DNA construct comprising a heterologous promoter functional in a plant cell and operably linked to: a) a polynucleotide that comprises a nucleotide sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity, or 100% identity to a sequence selected from the group consisting of SEQ ID NOs: 1-27 and 55; b) a DNA encoding RNA for suppressing the expression of a target mRNA transcribed from a polynucleotide having a nucleic acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity, or 100% identity to a sequence selected from the group consisting of SEQ ID NOs: 6 and 55-60; c) a polynucleotide that encodes a polypeptide having an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity, or 100% identity to a sequence selected from the group consisting of SEQ ID NOs: 28-54, 61, 79-96; or d) a DNA encoding RNA for suppressing the expression of a target protein having an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity, or 100% identity to a sequence selected from the group consisting of SEQ ID NOs: 33, 61-66, 82, and 96.
7. The plant of claim 6, wherein said plant has an altered phenotype or an enhanced trait as compared to a control plant.
8. The plant of claim 6, wherein said plant is a progeny, a propagule, or a field crop.
9. The plant of claim 6, wherein said plant is a field crop selected from the group consisting of corn, soybean, cotton, canola, rice, barley, oat, wheat, turf grass, alfalfa, sugar beet, sunflower, quinoa and sugar cane.
10. The plant of claim 6, wherein said plant is a propagule selected from the group consisting of cell, pollen, ovule, flower, embryo, leaf, root, stem, shoot, meristem, grain and seed.
11. The plant of claim 7, wherein said enhanced trait is selected from the group consisting of increased yield, increased nitrogen use efficiency, and increased water use efficiency as compared to a control plant.
12. The plant of claim 7, wherein said phenotype is selected from the group consisting of anthocyanin content, biomass, canopy area, chlorophyll content, plant height, water applied, water content and water use efficiency.
13. A method for increasing yield, increasing nitrogen use efficiency, or increasing water use efficiency in a plant comprising producing a plant comprising a recombinant DNA construct comprising a heterologous promoter functional in a plant cell and operably linked to: a) a polynucleotide that comprises a nucleotide sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity, or 100% identity to a sequence selected from the group consisting of SEQ ID NOs: 1-27 and 55; b) a DNA encoding RNA for suppressing the expression of a target mRNA transcribed from a polynucleotide having a nucleic acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity, or 100% identity to a sequence selected from the group consisting of SEQ ID NOs: 6 and 55-60; c) a polynucleotide that encodes a polypeptide having an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity, or 100% identity to a sequence selected from the group consisting of SEQ ID NOs: 28-54, 61, 79-96; or d) a DNA encoding RNA for suppressing the expression of a target protein having an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity, or 100% identity to a sequence selected from the group consisting of SEQ ID NOs: 33, 61-66, 82, and 96.
14. The method of claim 13 wherein said plant is produced by transforming a plant cell or tissue with said recombinant DNA construct and regenerating a plant from said cell or tissue containing said recombinant DNA construct.
15. The method of claim 13 comprising producing said plant by crossing said plant through breeding with: a) itself; b) a second plant from the same plant line; c) a wild type plant; or d) a second plant from a different line of plants to produce a seed, growing said seed to produce a plurality of progeny plants; and selecting a progeny plant with increased yield, increased nitrogen use efficiency, or increased water use efficiency as compared to a control plant.
Description:
CROSS REFERENCE TO RELATED APPLICATION
[0001] This application claims the benefit under 35USC .sctn. 119(e) of U.S. provisional application Ser. No. 62/086,918 filed on Dec. 3, 2014 herein incorporated by reference in its entirety.
INCORPORATION OF SEQUENCE LISTING
[0002] The sequence listing file named "60803WO0000_ST25.txt", which is 270 kilobytes (measured in MS-WINDOWS) and was created on Dec. 2, 2014, is filed herewith and incorporated herein by reference in its entirety.
FIELD OF THE INVENTION
[0003] Disclosed herein are recombinant DNA constructs, plants having enhanced traits such as increased yield, increased nitrogen use efficiency and increased water use efficiency; propagules, progenies and field crops of such plants; and methods of making and using such plants. Also disclosed are methods of producing seed from such plants, growing such seed and/or selecting progeny plants with enhanced traits.
SUMMARY OF THE INVENTION
[0004] In one aspect, the disclosure provides a recombinant DNA construct comprising a heterologous promoter functional in a plant cell and operably linked to:
[0005] a) a polynucleotide that comprises a nucleotide sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity, or 100% identity to a sequence selected from the group consisting of SEQ ID NOs: 1-27 and 55;
[0006] b) a DNA encoding RNA for suppressing the expression of a target mRNA transcribed from a polynucleotide having a nucleic acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity, or 100% identity to a sequence selected from the group consisting of SEQ ID NOs: 6 and 55-60;
[0007] c) a polynucleotide that encodes a polypeptide having an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity, or 100% identity to a sequence selected from the group consisting of SEQ ID NOs: 28-54, 61, 79-96; or
[0008] d) a DNA encoding RNA for suppressing the expression of a target protein having an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity, or 100% identity to a sequence selected from the group consisting of SEQ ID NOs: 33, 61-66, 82, and 96.
[0009] In another aspect, the disclosure provides a suppression recombinant DNA construct that transcribes into a double-stranded RNA, an antisense RNA, a miRNA or a ta-siRNA.
[0010] In another aspect, the disclosure provides a suppression recombinant DNA construct that transcribes into a miRNA precursor that produces a mature miRNA having a nucleic acid sequence with at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to a fragment of at least 19, 20, 21, 22, 23, 24, 25, 26 or 27 consecutive nucleotides of a sequence selected from the group consisting of SEQ ID NOs: 6 and 55-60.
[0011] In another aspect, the disclosure provides a suppression recombinant DNA construct that transcribes into a miRNA precursor that produces a mature miRNA having a nucleic acid sequence with at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% complimentarity to a fragment of at least 19, 20, 21, 22, 23, 24, 25, 26 or 27 consecutive nucleotides of a sequence selected from the group consisting of SEQ ID NOs: 6 and 55-60.
[0012] In another aspect, the disclosure provides a suppression recombinant DNA construct that transcribes into a miRNA precursor that produces a mature miRNA having a nucleic acid sequence with 100% identity or 100% complementarity to a fragment of 21 consecutive nucleotides of a sequence selected from the group consisting of SEQ ID NOs: 6 and 55-60.
[0013] In another aspect, the disclosure provides a suppression recombinant DNA construct comprising a sequence selected from the group consisting of SEQ ID NOs: 67-72.
[0014] In another aspect, the disclosure provides a plant comprising a recombinant DNA construct comprising a heterologous promoter functional in a plant cell and operably linked to:
[0015] a) a polynucleotide that comprises a nucleotide sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity, or 100% identity to a sequence selected from the group consisting of SEQ ID NOs: 1-27 and 55;
[0016] b) a DNA encoding RNA for suppressing the expression of a target mRNA transcribed from a polynucleotide having a nucleic acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity, or 100% identity to a sequence selected from the group consisting of SEQ ID NOs: 6 and 55-60;
[0017] c) a polynucleotide that encodes a polypeptide having an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity, or 100% identity to a sequence selected from the group consisting of SEQ ID NOs: 28-54, 61, 79-96; or
[0018] d) a DNA encoding RNA for suppressing the expression of a target protein having an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity, or 100% identity to a sequence selected from the group consisting of SEQ ID NOs: 33, 61-66, 82, and 96.
[0019] In another aspect, the disclosure provides a plant comprising a recombinant DNA construct of the present disclosure, and having at least one altered phenotype or at least one enhanced trait as compared to a control plant. Such phenotype is characterized or measured by anthocyanin content, biomass, canopy area, chlorophyll content, plant height, water applied, water content or water use efficiency. Such enhanced trait is increased yield, increased nitrogen use efficiency, or increased water use efficiency.
[0020] In another aspect, the disclosure provides a plant comprising a recombinant DNA construct of the present disclosure, wherein the plant is a progeny, a propagule, or a field crop.
[0021] In another aspect, the disclosure provides a field crop comprising a recombinant DNA construct of the present disclosure, wherein the field crop is selected from the group consisting of corn, soybean, cotton, canola, rice, barley, oat, wheat, turf grass, alfalfa, sugar beet, sunflower, quinoa and sugar cane.
[0022] In another aspect, the disclosure provides a propagule comprising a recombinant DNA construct the present disclosure, wherein the propagule is selected from the group consisting of cell, pollen, ovule, flower, embryo, leaf, root, stem, shoot, meristem, grain and seed.
[0023] In another aspect, the disclosure provides a plant comprising a recombinant DNA construct of the present disclosure, wherein the plant is a monocot plant or is a member of the family Poaceae, wheat plant, maize plant, sweet corn plant, rice plant, wild rice plant, barley plant, rye, millet plant, sorghum plant, sugar cane plant, turfgrass plant, bamboo plant, oat plant, brome-grass plant, Miscanthus plant, pampas grass plant, switchgrass (Panicum) plant, and/or teosinte plant, or is a member of the family Alliaceae, onion plant, leek plant, garlic plant; or wherein the plant is a dicot plant or is a member of the family Amaranthaceae, spinach plant, quinoa plant, a member of the family Anacardiaceae, mango plant, a member of the family Asteraceae, sunflower plant, endive plant, lettuce plant, artichoke plant, a member of the family Brassicaceae, Arabidopsis thaliana plant, rape plant, oilseed rape plant, broccoli plant, Brussels sprouts plant, cabbage plant, canola plant, cauliflower plant, kohlrabi plant, turnip plant, radish plant, a member of the family Bromeliaceae, pineapple plant, a member of the family Caricaceae, papaya plant, a member of the family Chenopodiaceae, beet plant, a member of the family Curcurbitaceae, melon plant, cantaloupe plant, squash plant, watermelon plant, honeydew plant, cucumber plant, pumpkin plant, a member of the family Dioscoreaceae, yam plant, a member of the family Ericaceae, blueberry plant, a member of the family Euphorbiaceae, cassava plant, a member of the family Fabaceae, alfalfa plant, clover plant, peanut plant, a member of the family Grossulariaceae, currant plant, a member of the family Juglandaceae, walnut plant, a member of the family Lamiaceae, mint plant, a member of the family Lauraceae, avocado plant, a member of the family Leguminosae, soybean plant, bean plant, pea plant, a member of the family Malvaceae, cotton plant, a member of the family Marantaceae, arrowroot plant, a member of the family Myrtaceae, guava plant, eucalyptus plant, a member of the family Rosaceae, peach plant, apple plant, cherry plant, plum plant, pear plant, prune plant, blackberry plant, raspberry plant, strawberry plant, a member of the family Rubiaceae, coffee plant, a member of the family Rutaceae, citrus plant, orange plant, lemon plant, grapefruit plant, tangerine plant, a member of the family Salicaceae, poplar plant, willow plant, a member of the family Solanaceae, potato plant, sweet potato plant, tomato plant, Capsicum plant, tobacco plant, tomatillo plant, eggplant plant, Atropa belladona plant, Datura stramonium plant, a member of the family Vitaceae, grape plant, a member of the family Umbelliferae, carrot plant, or a member of the family Musaceae, banana plant; or wherein the plant is a member of the family Pinaceae, cedar plant, fir plant, hemlock plant, larch plant, pine plant, or spruce plant.
[0024] In another aspect, the disclosure provides a method for increasing yield, increasing nitrogen use efficiency, or increasing water use efficiency in a plant comprising producing a plant comprising a recombinant DNA construct comprising a heterologous promoter functional in a plant cell and operably linked to:
a) a polynucleotide that comprises a nucleotide sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity, or 100% identity to a sequence selected from the group consisting of SEQ ID NOs: 1-27 and 55; b) a DNA encoding RNA for suppressing the expression of a target mRNA transcribed from a polynucleotide having a nucleic acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity, or 100% identity to a sequence selected from the group consisting of SEQ ID NOs: 6 and 55-60; c) a polynucleotide that encodes a polypeptide having an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity, or 100% identity to a sequence selected from the group consisting of SEQ ID NOs: 28-54, 61, 79-96; or d) a DNA encoding RNA for suppressing the expression of a target protein having an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity, or 100% identity to a sequence selected from the group consisting of SEQ ID NOs: 33, 61-66, 82, and 96.
[0025] In another aspect, the disclosure provides a method for producing a plant by transforming a plant cell or tissue with the recombinant DNA construct of the present disclosure and regenerating a plant from said cell or tissue containing said recombinant DNA construct. In another aspect, the disclosure provides a method for producing a plant by crossing said plant through breeding with:
[0026] a) itself;
[0027] b) a second plant from the same plant line;
[0028] c) a wild type plant; or
[0029] d) a second plant from a different line of plants to produce a seed, growing said seed to produce a plurality of progeny plants; and selecting a progeny plant with increased yield, increased nitrogen use efficiency, or increased water use efficiency as compared to a control plant.
DETAILED DESCRIPTION OF THE INVENTION
[0030] In the attached sequence listing:
[0031] SEQ ID NOs 1 to 27 are nucleotide sequences of the coding strand of the DNA used in the recombinant DNA constructs imparting an enhanced trait in plants, each representing a coding sequence for a protein.
[0032] SEQ ID NOs 28 to 54 are amino acid sequences of the cognate proteins of the DNA molecules with nucleotide sequences of SEQ ID NOs 1 to 27 respectively in the same order.
[0033] SEQ ID NOs: 55 to 60 are nucleotide sequences, each representing a coding sequence of a suppression target gene.
[0034] SEQ ID NOs 61 to 66 are amino acid sequences of the cognate proteins of the DNA molecules with nucleotide sequences of SEQ ID NOs 55 to 60 respectively in the same order.
[0035] SEQ ID NOs 67 to 72 are nucleotide sequences of DNA molecules used in the recombinant DNA constructs imparting an enhanced trait or altered phenotype in plants, each representing an engineered miRNA precursor sequence.
[0036] SEQ ID NOs: 73 to 78 are nucleotide sequences of the target recognition sites of the engineered miRNA precursors with nucleotide sequences of SEQ ID NOs 67 to 72 respectively in the same order.
[0037] SEQ ID NOs 79 to 96 are amino acid sequences of proteins homologous to the proteins with amino acid sequences of SEQ ID NOs 28 to 54, and 61 to 66.
[0038] SEQ ID NOs 97 to 100 are nucleotide sequences of DNA molecules used in the recombinant DNA constructs imparting an enhanced trait or altered phenotype in plants, each representing a promoter with a specific expression pattern.
[0039] SEQ ID NOs 101 to 104 are nucleotide sequences of variants of a rice MIR gene.
[0040] Unless otherwise stated, nucleic acid sequences in the text of this specification are given, when read from left to right, in the 5' to 3' direction. One of skill in the art would be aware that a given DNA sequence is understood to define a corresponding RNA sequence which is identical to the DNA sequence except for replacement of the thymine (T) nucleotide of the DNA with uracil (IU) nucleotide. Thus, providing a specific DNA sequence is understood to define the exact RNA equivalent. A given first polynucleotide sequence, whether DNA or RNA, further defines the sequence of its exact complement (which can be DNA or RNA), i. e., a second polynucleotide that hybridizes perfectly to the first polynucleotide by forming Watson-Crick base-pairs. By "essentially identical" or "essentially complementary" to a target gene or a fragment of a target gene is meant that a polynucleotide strand (or at least one strand of a double-stranded polynucleotide) is designed to hybridize (generally under physiological conditions such as those found in a living plant or animal cell) to a target gene or to a fragment of a target gene or to the transcript of the target gene or the fragment of a target gene; one of skill in the art would understand that such hybridization does not necessarily require 100% sequence identity or complementarity. A first nucleic acid sequence is "operably" connected or "linked" with a second nucleic acid sequence when the first nucleic acid sequence is placed in a functional relationship with the second nucleic acid sequence. For example, a promoter sequence is "operably linked" to DNA if the promoter provides for transcription or expression of the DNA. Generally, operably linked DNA sequences are contiguous.
[0041] As used herein, the term "expression" refers to the production of a polynucleotide or a protein by a plant, plant cell or plant tissue which can give rise to an altered phenotype or enhanced trait. Expression can also refer to the process by which information from a gene is used in the synthesis of functional gene products, which may include but are not limited to other polynucleotides or proteins which may serve, e.g., an enzymatic, structural or regulatory function. Gene products having a regulatory function include but are not limited to elements that affect the occurrence or level of transcription or translation of a target protein. In some cases, the expression product is a non-coding functional RNA.
[0042] "Modulation" of expression refers to the process of effecting either overexpression or suppression of a polynucleotide or a protein.
[0043] The term "suppression" as used herein refers to a lower expression level of a target polynucleotide or target protein in a plant, plant cell or plant tissue, as compared to the expression in a wild-type or control plant, cell or tissue, at any developmental or temporal stage for the gene. The term "target protein" as used in the context of suppression refers to a protein which is suppressed; similarly, "target mRNA" refers to a polynucleotide which can be suppressed or, once expressed, degraded so as to result in suppression of the target protein it encodes. The term "target gene" as used in the context of suppression refers to either "target protein" or "target mRNA". In alternate non-limiting embodiments, the target protein or target polynucleotide is one the suppression of which can give rise to an enhanced trait or altered phenotype directly or indirectly. In one exemplary embodiment, the target protein is one which can indirectly increase or decrease the expression of one or more other proteins, the increased or decreased expression, respectively, of which is associated with an enhanced trait or an altered phenotype. In another exemplary embodiment, the target protein can bind to one or more other proteins associated with an altered phenotype or enhanced trait to enhance or inhibit their function and thereby affect the altered phenotype or enhanced trait indirectly.
[0044] Suppression can be applied using numerous approaches. Non limiting examples include: suppressing an endogenous gene(s) or a subset of genes in a pathway, suppressing one or more mutation that has resulted in decreased activity of a protein, suppressing the production of an inhibitory agent, to elevate, reducing or eliminating the level of substrate that an enzyme requires for activity, producing a new protein, activating a normally silent gene; or accumulating a product that does not normally increase under natural conditions.
[0045] Conversely, the term "overexpression" as used herein refers to a greater expression level of a polynucleotide or a protein in a plant, plant cell or plant tissue, compared to expression in a wild-type plant, cell or tissue, at any developmental or temporal stage for the gene. Overexpression can take place in plant cells normally lacking expression of polypeptides functionally equivalent or identical to the present polypeptides. Overexpression can also occur in plant cells where endogenous expression of the present polypeptides or functionally equivalent molecules normally occurs, but such normal expression is at a lower level. Overexpression thus results in a greater than normal production, or "overproduction" of the polypeptide in the plant, cell or tissue.
[0046] The term "target protein" as used herein in the context of overexpression refers to a protein which is overexpressed; "target mRNA" refers to an mRNA which encodes and is translated to produce the target protein, which can also be overexpressed. The term "target gene" as used in the context of overexpression refers to either "target protein" or "target mRNA". In alternative embodiments, the target protein can effect an enhanced trait or altered phenotype directly or indirectly. In the latter case it may do so, for example, by affecting the expression, function or substrate available to one or more other proteins. In an exemplary embodiment, the target protein can bind to one or more other proteins associated with an altered phenotype or enhanced trait to enhance or inhibit their function.
[0047] Overexpression can be achieved using numerous approaches. In one embodiment, overexpression can be achieved by placing the DNA sequence encoding one or more polynucleotides or polypeptides under the control of a promoter, examples of which include but are not limited to endogenous promoters, heterologous promoters, inducible promoters and tissue specific promoters. In one exemplary embodiment, the promoter is a constitutive promoter, for example, the cauliflower mosaic virus 35S transcription initiation region. Thus, depending on the promoter used, overexpression can occur throughout a plant, in specific tissues of the plant, or in the presence or absence of different inducing or inducible agents, such as hormones or environmental signals.
[0048] Gene Suppression Elements: The gene suppression element can be transcribable DNA of any suitable length, and generally includes at least about 19 to about 27 nucleotides (for example 19, 20, 21, 22, 23, or 24 nucleotides) for every target gene that the recombinant DNA construct is intended to suppress. In many embodiments the gene suppression element includes more than 23 nucleotides (for example, more than about 30, about 50, about 100, about 200, about 300, about 500, about 1000, about 1500, about 2000, about 3000, about 4000, or about 5000 nucleotides) for every target gene that the recombinant DNA construct is intended to suppress.
[0049] Suitable gene suppression elements useful in the recombinant DNA constructs of the invention include at least one element (and, in some embodiments, multiple elements) selected from the group consisting of:
(a) DNA that includes at least one anti-sense DNA segment that is anti-sense to at least one segment of the at least one first target gene; (b) DNA that includes multiple copies of at least one anti-sense DNA segment that is anti-sense to at least one segment of the at least one first target gene; (c) DNA that includes at least one sense DNA segment that is at least one segment of the at least one first target gene; (d) DNA that includes multiple copies of at least one sense DNA segment that is at least one segment of the at least one first target gene: (e) DNA that transcribes to RNA for suppressing the at least one first target gene by forming double-stranded RNA and includes at least one anti-sense DNA segment that is anti-sense to at least one segment of the at least one target gene and at least one sense DNA segment that is at least one segment of the at least one first target gene; (f) DNA that transcribes to RNA for suppressing the at least one first target gene by forming a single double-stranded RNA and includes multiple serial anti-sense DNA segments that are anti-sense to at least one segment of the at least one first target gene and multiple serial sense DNA segments that are at least one segment of the at least one first target gene; (g) DNA that transcribes to RNA for suppressing the at least one first target gene by forming multiple double strands of RNA and includes multiple anti-sense DNA segments that are anti-sense to at least one segment of the at least one first target gene and multiple sense DNA segments that are at least one segment of the at least one first target gene, and wherein said multiple anti-sense DNA segments and the multiple sense DNA segments are arranged in a series of inverted repeats; (h) DNA that includes nucleotides derived from a miRNA, preferably a plant miRNA; (i) DNA that includes nucleotides of a siRNA; j) DNA that transcribes to an RNA aptamer capable of binding to a ligand; and (k) DNA that transcribes to an RNA aptamer capable of binding to a ligand, and DNA that transcribes to regulatory RNA capable of regulating expression of the first target gene, wherein the regulation is dependent on the conformation of the regulatory RNA, and the conformation of the regulatory RNA is allosterically affected by the binding state of the RNA aptamer.
[0050] Any of these gene suppression elements, whether transcribing to a single double-stranded RNA or to multiple double-stranded RNAs, can be designed to suppress more than one target gene, including, for example, more than one allele of a target gene, multiple target genes (or multiple segments of at least one target gene) from a single species, or target genes from different species.
[0051] Anti-Sense DNA Segments: In one embodiment, the at least one anti-sense DNA segment that is anti-sense to at least one segment of the at least one first target gene includes DNA sequence that is anti-sense or complementary to at least a segment of the at least one first target gene, and can include multiple anti-sense DNA segments, that is, multiple copies of at least one anti-sense DNA segment that is anti-sense to at least one segment of the at least one first target gene. Multiple anti-sense DNA segments can include DNA sequence that is anti-sense or complementary to multiple segments of the at least one first target gene, or to multiple copies of a segment of the at least one first target gene, or to segments of multiple first target genes, or to any combination of these. Multiple anti-sense DNA segments can be fused into a chimera, e.g., including DNA sequences that are anti-sense to multiple segments of one or more first target genes and fused together.
[0052] The anti-sense DNA sequence that is anti-sense or complementary to (that is, can form Watson-Crick base-pairs with) at least a segment of the at least one first target gene has at least about 80%, or at least about 85%, or at least about 90%, or at least about 95% complementarity to at least a segment of the at least one first target gene. In one embodiment, the DNA sequence that is anti-sense or complementary to at least a segment of the at least one first target gene has between about 95% to about 100% complementarity to at least a segment of the at least one first target gene. Where the at least one anti-sense DNA segment includes multiple anti-sense DNA segments, the degree of complementarity can be, but need not be, identical for all of the multiple anti-sense DNA segments.
[0053] Sense DNA Segments: In another embodiment, the at least one sense DNA segment that is at least one segment of the at least one first target gene includes DNA sequence that corresponds to (that is, has a sequence that is identical or substantially identical to) at least a segment of the at least one first target gene, and can include multiple sense DNA segments, that is, multiple copies of at least one sense DNA segment that corresponds to (that is, has the nucleotide sequence of) at least one segment of the at least one first target gene. Multiple sense DNA segments can include DNA sequence that is or that corresponds to multiple segments of the at least one first target gene, or to multiple copies of a segment of the at least one first target gene, or to segments of multiple first target genes, or to any combination of these. Multiple sense DNA segments can be fused into a chimera, that is, can include DNA sequences corresponding to multiple segments of one or more first target genes and fused together.
[0054] The sense DNA sequence that corresponds to at least a segment of the target gene has at least about 80%, or at least about 85%, or at least about 90%, or at least about 95% sequence identity to at least a segment of the target gene. In one embodiment, the DNA sequence that corresponds to at least a segment of the target gene has between about 95% to about 100% sequence identity to at least a segment of the target gene. Where the at least one sense DNA segment includes multiple sense DNA segments, the degree of sequence identity can be, but need not be, identical for all of the multiple sense DNA segments.
[0055] Multiple Copies: Where the gene suppression element includes multiple copies of anti-sense or multiple copies of sense DNA sequence, these multiple copies can be arranged serially in tandem repeats. In some embodiments, these multiple copies can be arranged serially end-to-end, that is, in directly connected tandem repeats. In some embodiments, these multiple copies can be arranged serially in interrupted tandem repeats, where one or more spacer DNA segment can be located adjacent to one or more of the multiple copies. Tandem repeats, whether directly connected or interrupted or a combination of both, can include multiple copies of a single anti-sense or multiple copies of a single sense DNA sequence in a serial arrangement or can include multiple copies of more than one anti-sense DNA sequence or of more than one sense DNA sequence in a serial arrangement.
[0056] Double-stranded RNA: In those embodiments wherein the gene suppression element includes either at least one anti-sense DNA segment that is anti-sense to at least one segment of the at least one target gene or at least one sense DNA segment that is at least one segment of the at least one target gene, RNA transcribed from either the at least one anti-sense or at least one sense DNA may become double-stranded by the action of an RNA-dependent RNA polymerase. See, for example, U.S. Pat. No. 5,283,184, which is incorporated by reference herein.
[0057] In yet other embodiments, the gene suppression element can include DNA that transcribes to RNA for suppressing the at least one first target gene by forming double-stranded RNA and includes at least one anti-sense DNA segment that is anti-sense to at least one segment of the at least one target gene (as described above under the heading "Anti-sense DNA Segments") and at least one sense DNA segment that is at least one segment of the at least one first target gene (as described above under the heading "Sense DNA Segments"). Such a gene suppression element can further include spacer DNA segments. Each at least one anti-sense DNA segment is complementary to at least part of a sense DNA segment in order to permit formation of double-stranded RNA by intramolecular hybridization of the at least one anti-sense DNA segment and the at least one sense DNA segment. Such complementarity between an anti-sense DNA segment and a sense DNA segment can be, but need not be, 100% complementarity; in some embodiments, this complementarity can be preferably at least about 80%, or at least about 85%, or at least about 90%, or at least about 95% complementarity.
[0058] The double-stranded RNA can be in the form of a single dsRNA "stem" (region of base-pairing between sense and anti-sense strands), or can have multiple dsRNA "stems". In one embodiment, the gene suppression element can include DNA that transcribes to RNA for suppressing the at least one first target gene by forming essentially a single double-stranded RNA and includes multiple serial anti-sense DNA segments that are anti-sense to at least one segment of the at least one first target gene and multiple serial sense DNA segments that are at least one segment of the at least one first target gene; the multiple serial anti-sense and multiple serial sense segments can form a single double-stranded RNA "stem" or multiple "stems" in a serial arrangement (with or without non-base paired spacer DNA separating the multiple "stems"). In another embodiment, the gene suppression element includes DNA that transcribes to RNA for suppressing the at least one first target gene by forming multiple dsRNA "stems" of RNA and includes multiple anti-sense DNA segments that are anti-sense to at least one segment of the at least one first target gene and multiple sense DNA segments that are at least one segment of the at least one first target gene, and wherein said multiple anti-sense DNA segments and the multiple sense DNA segments are arranged in a series of dsRNA "stems" (such as, but not limited to "inverted repeats"). Such multiple dsRNA "stems" can further be arranged in series or clusters to form tandem inverted repeats, or structures resembling "hammerhead" or "cloverleaf" shapes. Any of these gene suppression elements can further include spacer DNA segments found within a dsRNA "stem" (for example, as a spacer between multiple anti-sense or sense DNA segments or as a spacer between a base-pairing anti-sense DNA segment and a sense DNA segment) or outside of a double-stranded RNA "stem" (for example, as a loop region separating a pair of inverted repeats). In cases where base-pairing anti-sense and sense DNA segment are of unequal length, the longer segment can act as a spacer.
[0059] miRNAs: In a further embodiment, the gene suppression element can include DNA that includes nucleotides derived from a miRNA (microRNA), that is, a DNA sequence that corresponds to a miRNA native to a virus or a eukaryote of interest (including plants and animals, especially invertebrates), or a DNA sequence derived from such a native miRNA but modified to include nucleotide sequences that do not correspond to the native miRNA. While miRNAs have not to date been reported in fungi, fungal miRNAs, should they exist, are also suitable for use in the invention. An embodiment includes a gene suppression element containing DNA that includes nucleotides derived from a viral or plant miRNA.
[0060] In a non-limiting example, the nucleotides derived from a miRNA can include DNA that includes nucleotides corresponding to the loop region of a native miRNA and nucleotides that are selected from a target gene sequence. In another non-limiting example, the nucleotides derived from a miRNA can include DNA derived from a miRNA precursor sequence, such as a native pri-miRNA or pre-miRNA sequence, or nucleotides corresponding to the regions of a native miRNA and nucleotides that are selected from a target gene sequence number such that the overall structure (e.g., the placement of mismatches in the stem structure of the pre-miRNA) is preserved to permit the pre-miRNA to be processed into a mature miRNA. In yet another embodiment, the gene suppression element can include DNA that includes nucleotides derived from a miRNA and capable of inducing or guiding in-phase cleavage of an endogenous transcript into trans-acting siRNAs, as described by Allen et al. (2005) Cell, 121:207-221, which is incorporated by reference in its entirety herein. Thus, the DNA that includes nucleotides derived from a miRNA can include sequence naturally occurring in a miRNA or a miRNA precursor molecule, synthetic sequence, or both.
[0061] siRNAs: In yet another embodiment, the gene suppression element can include DNA that includes nucleotides of a small interfering RNA (siRNA). The siRNA can be one or more native siRNAs (such as siRNAs isolated from a non-transgenic eukaryote or from a transgenic eukaryote), or can be one or more DNA sequences predicted to have siRNA activity (such as by use of predictive tools known in the art, see, for example. Reynolds et al. (2004) Nature Biotechnol., 22:326-330, which is incorporated by reference in its entirety herein). Multiple native or predicted siRNA sequences can be joined in a chimeric siRNA sequence for gene suppression. Such a DNA that includes nucleotides of a siRNA includes at least 19 nucleotides, and in some embodiments includes at least 20, at least 21, at least 22, at least 23, or at least 24 nucleotides. In other embodiments, the DNA that includes nucleotides of a siRNA can contain substantially more than 21 nucleotides, for example, more than about 50, about 100, about 300, about 500, about 1000, about 3000, or about 5000 nucleotides or greater.
[0062] Engineered miRNAs and trans-acting siRNAs (ta-siRNAs) are useful for gene suppression with increased specificity. The invention provides recombinant DNA constructs, each including a transcribable engineered miRNA precursor designed to suppress a target sequence, wherein the transcribable engineered miRNA precursor is derived from the fold-back structure of a MIR gene, preferably a plant MIR sequence. These miRNA precursors are also useful for directing in-phase production of siRNAs (e.g., heterologous sequence designed to be processed in a trans-acting siRNA suppression mechanism in planta). The invention further provides a method to suppress expression of a target sequence in a plant cell, including transcribing in a plant cell a recombinant DNA including a transcribable engineered miRNA precursor designed to suppress a target sequence, wherein the transcribable engineered miRNA precursor is derived from the fold-back structure of a MIR gene, preferably a plant MIR sequence, whereby expression of the target sequence is suppressed relative to its expression in the absence of transcription of the recombinant DNA construct. In specifically claimed embodiments, the transcribable engineered miRNA precursor is derived from the fold-back structure of a rice MIR sequence selected from the group consisting of SEQ ID NOs. 101-104, and their complements.
[0063] The mature miRNAs produced, or predicted to be produced, from these miRNA precursors may be engineered for use in suppression of a target gene, e.g., in transcriptional suppression by the miRNA, or to direct in-phase production of siRNAs in a trans-acting siRNA suppression mechanism (see Allen et al. (2005) Cell, 121:207-221, Vaucheret (2005) Science STKE, 2005:pe43, and Yoshikawa et al. (2005) Genes Dev., 19:2164-2175, all of which are incorporated by reference herein). Plant miRNAs generally have near-perfect complementarity to their target sequences (see, for example. Llave et al. (2002) Science. 297:2053-2056, Rhoades et al. (2002) Cell, 110:513-520, Jones-Rhoades and Bartel (2004) Mol. Cell, 14:787-799, all of which are incorporated by reference herein). Thus, the mature miRNAs can be engineered to serve as sequences useful for gene suppression of a target sequence, by replacing nucleotides of the mature miRNA sequence with nucleotides of the sequence that is targeted for suppression; see, for example, methods disclosed by Parizotto et al. (2004) Genes Dev., 18:2237-2242 and especially U.S. Patent Application Publications US2004/0053411A1, US2004/0268441A1, US2005/0144669, and US2005/0037988 all of which are incorporated by reference herein. When engineering a novel miRNA to target a specific sequence, one strategy is to select within the target sequence a region with sequence that is as similar as possible to the native miRNA sequence. Alternatively, the native miRNA sequence can be replaced with a region of the target sequence, preferably a region that meets structural and thermodynamic criteria believed to be important for miRNA function (see, for example, U.S. Patent Application Publication US2005/0037988). Sequences are preferably engineered such that the number and placement of mismatches in the stem structure of the fold-back region or pre-miRNA is preserved. Thus, an engineered miRNA or engineered miRNA precursor can be derived from any of the mature miRNA sequences, or their corresponding miRNA precursors (including the fold-back portions of the corresponding MIR genes) disclosed herein. The engineered miRNA precursor can be cloned and expressed (transiently or stably) in a plant cell or tissue or intact plant.
[0064] The construction and description of recombinant DNA constructs to modulate small non-coding RNA activities are disclosed in US Patent Application Publication US 2009/0070898 A1, US2011/0296555 A1, US2011/0035839 A1, all of which are incorporated herein by reference in their entirety. In particular, with respect to US2011/0035839 A1, see e.g., sections under the headings "Gene Suppression Elements" in paragraphs 122 to 135, and "Engineered Heterologous miRNA for Controlling Gene Expression in paragraphs 188 to 190.
[0065] As used herein a "plant" includes a whole plant, a transgenic plant, meristematic tissue, a shoot organ/structure (for example, leaf, stem and tuber), a root, a flower, a floral organ/structure (for example, a bract, a sepal, a petal, a stamen, a carpel, an anther and an ovule), a seed (including an embryo, endosperm, and a seed coat) and a fruit (the mature ovary), plant tissue (for example, vascular tissue, ground tissue, and the like) and a cell (for example, guard cell, egg cell, pollen, mesophyll cell, and the like), and progeny of same. The classes of plants that can be used in the disclosed methods are generally as broad as the classes of higher and lower plants amenable to transformation and breeding techniques, including angiosperms (monocotyledonous and dicotyledonous plants), gymnosperms, ferns, horsetails, psilophytes, lycophytes, bryophytes, and multicellular algae.
[0066] As used herein a "transgenic plant cell" means a plant cell that is transformed with stably-integrated, recombinant DNA, for example, by Agrobacterium-mediated transformation or by bombardment using microparticles coated with recombinant DNA or by other means. A plant cell of this disclosure can be an originally-transformed plant cell that exists as a microorganism or as a progeny plant cell that is regenerated into differentiated tissue, for example, into a transgenic plant with stably-integrated, recombinant DNA, or seed or pollen derived from a progeny transgenic plant.
[0067] As used herein a "control plant" means a plant that does not contain the recombinant DNA of the present disclosure that imparts an enhanced trait or altered phenotype. A control plant is used to identify and select a transgenic plant that has an enhanced trait or altered phenotype. A suitable control plant can be a non-transgenic plant of the parental line used to generate a transgenic plant, for example, a wild type plant devoid of a recombinant DNA. A suitable control plant can also be a transgenic plant that contains recombinant DNA that imparts other traits, for example, a transgenic plant having enhanced herbicide tolerance. A suitable control plant can in some cases be a progeny of a hemizygous transgenic plant line that does not contain the recombinant DNA, known as a negative segregant, or a negative isogenic line.
[0068] As used herein a "propagule" includes all products of meiosis and mitosis, including but not limited to, plant, seed and part of a plant able to propagate a new plant. Propagules include whole plants, cells, pollen, ovules, flowers, embryos, leaves, roots, stems, shoots, meristems, grains or seeds, or any plant part that is capable of growing into an entire plant. Propagule also includes graft where one portion of a plant is grafted to another portion of a different plant (even one of a different species) to create a living organism. Propagule also includes all plants and seeds produced by cloning or by bringing together meiotic products, or allowing meiotic products to come together to form an embryo or a fertilized egg (naturally or with human intervention).
[0069] As used herein a "progeny" includes any plant, seed, plant cell, and/or regenerable plant part comprising a recombinant DNA of the present disclosure derived from an ancestor plant. A progeny can be homozygous or heterozygous for the transgene. Progeny can be grown from seeds produced by a transgenic plant comprising a recombinant DNA of the present disclosure, and/or from seeds produced by a plant fertilized with pollen or ovule from a transgenic plant comprising a recombinant DNA of the present disclosure.
[0070] As used herein a "trait" is a physiological, morphological, biochemical, or physical characteristic of a plant or particular plant material or cell. In some instances, this characteristic is visible to the human eye, such as seed or plant size, or can be measured by biochemical techniques, such as detecting the protein, starch, certain metabolites, or oil content of seed or leaves, or by observation of a metabolic or physiological process, for example, by measuring tolerance to water deprivation or particular salt or sugar concentrations, or by the measurement of the expression level of a gene or genes, for example, by employing Northern analysis, RT-PCR, microarray gene expression assays, or reporter gene expression systems, or by agricultural observations such as hyperosmotic stress tolerance or yield. Any technique can be used to measure the amount of, comparative level of, or difference in any selected chemical compound or macromolecule in the transgenic plants, however.
[0071] As used herein an "enhanced trait" means a characteristic of a transgenic plant as a result of stable integration and expression of a recombinant DNA in the transgenic plant. Such traits include, but are not limited to, an enhanced agronomic trait characterized by enhanced plant morphology, physiology, growth and development, yield, nutritional enhancement, disease or pest resistance, or environmental or chemical tolerance. In some specific aspects of this disclosure an enhanced trait is selected from the group consisting of drought tolerance, increased water use efficiency, cold tolerance, increased nitrogen use efficiency and increased yield as shown in Tables 7 and 9, and altered phenotypes as shown in Tables 4-6. In another aspect of the disclosure the trait is increased yield under non-stress conditions or increased yield under environmental stress conditions. Stress conditions can include both biotic and abiotic stress, for example, drought, shade, fungal disease, viral disease, bacterial disease, insect infestation, nematode infestation, cold temperature exposure, heat exposure, osmotic stress, reduced nitrogen nutrient availability, reduced phosphorus nutrient availability and high plant density. "Yield" can be affected by many properties including without limitation, plant height, plant biomass, pod number, pod position on the plant, number of internodes, incidence of pod shatter, grain size, efficiency of nodulation and nitrogen fixation, efficiency of nutrient assimilation, resistance to biotic and abiotic stress, carbon assimilation, plant architecture, resistance to lodging, percent seed germination, seedling vigor, and juvenile traits. Yield can also be affected by efficiency of germination (including germination in stressed conditions), growth rate (including growth rate in stressed conditions), ear number, ear size, ear weight, seed number per ear or pod, seed size, composition of seed (starch, oil, protein) and characteristics of seed fill.
[0072] Also used herein, the term "trait modification" encompasses altering the naturally occurring trait by producing a detectable difference in a characteristic in a plant comprising a recombinant DNA of the present disclosure relative to a plant not comprising the recombinant DNA, such as a wild-type plant, or a negative segregant. In some cases, the trait modification can be evaluated quantitatively. For example, the trait modification can entail an increase or decrease, in an observed trait as compared to a control plant. It is known that there can be natural variations in a modified trait. Therefore, the trait modification observed entails a change of the normal distribution and magnitude of the trait in the plants as compared to a control plant.
[0073] The present disclosure relates to a plant with improved economically important characteristics, more specifically increased yield. More specifically the present disclosure relates to a plant comprising a polynucleotide of this disclosure, wherein the plant has increased yield as compared to a control plant. Many plants of this disclosure exhibited increased yield as compared to a control plant. In an embodiment, a plant of the present disclosure exhibited an improved trait that is related to yield, including but not limited to increased nitrogen use efficiency, increased nitrogen stress tolerance, increased water use efficiency and increased drought tolerance, as defined and discussed infra.
[0074] Yield can be defined as the measurable produce of economic value from a crop. Yield can be defined in the scope of quantity and/or quality. Yield can be directly dependent on several factors, for example, the number and size of organs, plant architecture (such as the number of branches, plant biomass, etc.), seed production and more. Root development, photosynthetic efficiency, nutrient uptake, stress tolerance, early vigor, delayed senescence and functional stay green phenotypes can be important factors in determining yield. Optimizing the above mentioned factors can therefore contribute to increasing crop yield.
[0075] Reference herein to an increase in yield-related traits can also be taken to mean an increase in biomass (weight) of one or more parts of a plant, which can include above ground and/or below ground (harvestable) plant parts. In particular, such harvestable parts are seeds, and performance of the methods of the disclosure results in plants with increased yield and in particular increased seed yield relative to the seed yield of suitable control plants. The term "yield" of a plant can relate to vegetative biomass (root and/or shoot biomass), to reproductive organs, and/or to propagules (such as seeds) of that plant.
[0076] Increased yield of a plant of the present disclosure can be measured in a number of ways, including test weight, seed number per plant, seed weight, seed number per unit area (for example, seeds, or weight of seeds, per acre), bushels per acre, tons per acre, or kilo per hectare. For example, corn yield can be measured as production of shelled corn kernels per unit of production area, for example in bushels per acre or metric tons per hectare. This is often also reported on a moisture adjusted basis, for example at 15.5 percent moisture. Increased yield can result from improved utilization of key biochemical compounds, such as nitrogen, phosphorous and carbohydrate, or from improved responses to environmental stresses, such as cold, heat, drought, salt, shade, high plant density, and attack by pests or pathogens. This disclosure can also be used to provide plants with improved growth and development, and ultimately increased yield, as the result of modified expression of plant growth regulators or modification of cell cycle or photosynthesis pathways. Also of interest is the generation of plants that demonstrate increased yield with respect to a seed component that may or may not correspond to an increase in overall plant yield.
[0077] In an embodiment, "alfalfa yield" can also be measured in forage yield, the amount of above ground biomass at harvest. Factors leading contributing to increased biomass include increased vegetative growth, branches, nodes and internodes, leaf area, and leaf area index.
[0078] In another embodiment, "canola yield" can also be measured in pod number, number of pods per plant, number of pods per node, number of internodes, incidence of pod shatter, seeds per silique, seed weight per silique, improved seed, oil, or protein composition.
[0079] Additionally, "corn or maize yield" can also be measured as production of shelled corn kernels per unit of production area, ears per acre, number of kernel rows per ear and number of kernels per row, kernel number or weight per ear, weight per kernel, ear number, ear weight, fresh or dry ear biomass (weight).
[0080] In yet another embodiment. "cotton yield" can be measured as bolls per plant, size of bolls, fiber quality, seed cotton yield in g/plant, seed cotton yield in lb/acre, lint yield in lb/acre, and number of bales.
[0081] Specific embodiment for "rice yield" can also include panicles per hill, grain per hill, and filled grains per panicle.
[0082] Still further embodiment for "soybean yield" can also include pods per plant, pods per acre, seeds per plant, seeds per pod, weight per seed, weight per pod, pods per node, number of nodes, and the number of internodes per plant.
[0083] In still further embodiment, "sugarcane yield" can be measured as cane yield (tons per acre; kg/hectare), total recoverable sugar (pounds per ton), and sugar yield (tons/acre).
[0084] In yet still further embodiment, "wheat yield" can include: cereal per unit area, grain number, grain weight, grain size, grains per head, seeds per head, seeds per plant, heads per acre, number of viable tillers per plant, composition of seed (for example, carbohydrates, starch, oil, and protein) and characteristics of seed fill.
[0085] The terms "yield", "seed yield" are defined above for a number of core crops. The terms "increased", "improved", "enhanced" are interchangeable and are defined herein.
[0086] In another embodiment, the present disclosure provides a method for the production of plants having increased yield; performance of the method gives plants increased yield. "Increased yield" can manifest as one or more of the following: (i) increased plant biomass (weight) of one or more parts of a plant, particularly aboveground (harvestable) parts, of a plant, increased root biomass (increased number of roots, increased root thickness, increased root length) or increased biomass of any other harvestable part; or (ii) increased early vigor, defined herein as an improved seedling aboveground area approximately three weeks post-germination. "Early vigor" refers to active healthy plant growth especially during early stages of plant growth, and can result from increased plant fitness due to, for example, the plants being better adapted to their environment (for example, optimizing the use of energy resources, uptake of nutrients and partitioning carbon allocation between shoot and root). Early vigor in corn, for example, is a combination of the ability of corn seeds to germinate and emerge after planting and the ability of the young corn plants to grow and develop after emergence. Plants having early vigor also show increased seedling survival and better establishment of the crop, which often results in highly uniform fields with the majority of the plants reaching the various stages of development at substantially the same time, which often results in increased yield. Therefore early vigor can be determined by measuring various factors, such as kernel weight, percentage germination, percentage emergence, seedling growth, seedling height, root length, root and shoot biomass, canopy size and color and others.
[0087] Further, increased yield can also manifest as (iii) increased total seed yield, which may result from one or more of an increase in seed biomass (seed weight) due to an increase in the seed weight on a per plant and/or on an individual seed basis an increased number of panicles per plant; an increased number of pods; an increased number of nodes; an increased number of flowers ("florets") per panicle/plant; increased seed fill rate; an increased number of filled seeds; increased seed size (length, width, area, perimeter), which can also influence the composition of seeds; and/or increased seed volume, which can also influence the composition of seeds.
[0088] Increased yield can also (iv) result in modified architecture, or can occur because of modified plant architecture.
[0089] Increased yield can also manifest as (v) increased harvest index, which is expressed as a ratio of the yield of harvestable parts, such as seeds, over the total biomass
[0090] Increased yield can also manifest as (vi) increased kernel weight, which is extrapolated from the number of filled seeds counted and their total weight. An increased kernel weight can result from an increased seed size and/or seed weight, an increase in embryo size, increased endosperm size, aleurone and/or scutellum, or an increase with respect to other parts of the seed that result in increased kernel weight.
[0091] Increased yield can also manifest as (vii) increased ear biomass, which is the weight of the ear and can be represented on a per ear, per plant or per plot basis.
[0092] In one embodiment, increased yield can be increased seed yield, and is selected from one of the following: (i) increased seed weight; (ii) increased number of filled seeds; and (iii) increased harvest index.
[0093] The disclosure also extends to harvestable parts of a plant such as, but not limited to, seeds, leaves, fruits, flowers, bolls, stems, rhizomes, tubers and bulbs. The disclosure furthermore relates to products derived from a harvestable part of such a plant, such as dry pellets, powders, oil, fat and fatty acids, starch or proteins.
[0094] The present disclosure provides a method for increasing "yield" of a plant or "broad acre yield" of a plant or plant part defined as the harvestable plant parts per unit area, for example seeds, or weight of seeds, per acre, pounds per acre, bushels per acre, tones per acre, tons per acre, kilo per hectare.
[0095] This disclosure further provides a method of increasing yield in a plant by producing a plant comprising a polynucleic acid sequence of this disclosure where the plant can be crossed with itself, a second plant from the same plant line, a wild type plant, or a plant from a different line of plants to produce a seed. The seed of the resultant plant can be harvested from fertile plants and be used to grow progeny generations of plant(s) of this disclosure. In addition to direct transformation of a plant with a recombinant DNA construct, transgenic plants can be prepared by crossing a first plant having a stably integrated recombinant DNA construct with a second plant lacking the DNA. For example, recombinant DNA can be introduced into a first plant line that is amenable to transformation to produce a transgenic plant which can be crossed with a second plant line to introgress the recombinant DNA into the second plant line.
[0096] Selected transgenic plants transformed with a recombinant DNA construct and having the polynucleotide of this disclosure provides the enhanced trait of increased yield compared to a control plant. Use of genetic markers associated with the recombinant DNA can facilitate production of transgenic progeny that is homozygous for the desired recombinant DNA. Progeny plants carrying DNA for both parental traits can be back-crossed into a parent line multiple times, for example usually 6 to 8 generations, to produce a progeny plant with substantially the same genotype as the one reoccurring original transgenic parental line but having the recombinant DNA of the other transgenic parental line. The term "progeny" denotes the offspring of any generation of a parent plant prepared by the methods of this disclosure containing the recombinant polynucleotides as described herein.
[0097] As used herein "nitrogen use efficiency" refers to the processes which lead to an increase in the plant's yield, biomass, vigor, and growth rate per nitrogen unit applied. The processes can include the uptake, assimilation, accumulation, signaling, sensing, retranslocation (within the plant) and use of nitrogen by the plant.
[0098] As used herein "nitrogen limiting conditions" refers to growth conditions or environments that provide less than optimal amounts of nitrogen needed for adequate or successful plant metabolism, growth, reproductive success and/or viability.
[0099] As used herein the "increased nitrogen stress tolerance" refers to the ability of plants to grow, develop, or yield normally, or grow, develop, or yield faster or better when subjected to less than optimal amounts of available/applied nitrogen, or under nitrogen limiting conditions.
[0100] As used herein "increased nitrogen use efficiency" refers to the ability of plants to grow, develop, or yield faster or better than normal when subjected to the same amount of available/applied nitrogen as under normal or standard conditions; ability of plants to grow, develop, or yield normally, or grow, develop, or yield faster or better when subjected to less than optimal amounts of available/applied nitrogen, or under nitrogen limiting conditions.
[0101] Increased plant nitrogen use efficiency can be translated in the field into either harvesting similar quantities of yield, while supplying less nitrogen, or increased yield gained by supplying optimal/sufficient amounts of nitrogen. The increased nitrogen use efficiency can improve plant nitrogen stress tolerance, and can also improve crop quality and biochemical constituents of the seed such as protein yield and oil yield. The terms "increased nitrogen use efficiency", "enhanced nitrogen use efficiency", and "nitrogen stress tolerance" are used inter-changeably in the present disclosure to refer to plants with improved productivity under nitrogen limiting conditions.
[0102] As used herein "water use efficiency" refers to the amount of carbon dioxide assimilated by leaves per unit of water vapor transpired. It constitutes one of the most important traits controlling plant productivity in dry environments. "Drought tolerance" refers to the degree to which a plant is adapted to arid or drought conditions. The physiological responses of plants to a deficit of water include leaf wilting, a reduction in leaf area, leaf abscission, and the stimulation of root growth by directing nutrients to the underground parts of the plants. Plants are more susceptible to drought during flowering and seed development (the reproductive stages), as plant's resources are deviated to support root growth. In addition, abscisic acid (ABA), a plant stress hormone, induces the closure of leaf stomata (microscopic pores involved in gas exchange), thereby reducing water loss through transpiration, and decreasing the rate of photosynthesis. These responses improve the water-use efficiency of the plant on the short term. The terms "increased water use efficiency", "enhanced water use efficiency", and "increased drought tolerance" are used inter-changeably in the present disclosure to refer to plants with improved productivity under water-limiting conditions.
[0103] As used herein "increased water use efficiency" refers to the ability of plants to grow, develop, or yield faster or better than normal when subjected to the same amount of available/applied water as under normal or standard conditions; ability of plants to grow, develop, or yield normally, or grow, develop, or yield faster or better when subjected to reduced amounts of available/applied water (water input) or under conditions of water stress or water deficit stress.
[0104] As used herein "increased drought tolerance" refers to the ability of plants to grow, develop, or yield normally, or grow, develop, or yield faster or better than normal when subjected to reduced amounts of available/applied water and/or under conditions of acute or chronic drought; ability of plants to grow, develop, or yield normally when subjected to reduced amounts of available/applied water (water input) or under conditions of water deficit stress or under conditions of acute or chronic drought.
[0105] As used herein "drought stress" refers to a period of dryness (acute or chronic/prolonged) that results in water deficit and subjects plants to stress and/or damage to plant tissues and/or negatively affects grain/crop yield; a period of dryness (acute or chronic/prolonged) that results in water deficit and/or higher temperatures and subjects plants to stress and/or damage to plant tissues and/or negatively affects grain/crop yield.
[0106] As used herein "water deficit" refers to the conditions or environments that provide less than optimal amounts of water needed for adequate/successful growth and development of plants.
[0107] As used herein "water stress" refers to the conditions or environments that provide improper (either less/insufficient or more/excessive) amounts of water than that needed for adequate/successful growth and development of plants/crops thereby subjecting the plants to stress and/or damage to plant tissues and/or negatively affecting grain/crop yield.
[0108] As used herein "water deficit stress" refers to the conditions or environments that provide less/insufficient amounts of water than that needed for adequate/successful growth and development of plants/crops thereby subjecting the plants to stress and/or damage to plant tissues and/or negatively affecting grain yield.
[0109] As used herein a "polynucleotide" is a nucleic acid molecule comprising a plurality of polymerized nucleotides. A polynucleotide may be referred to as a nucleic acid, a oligonucleotide, or any fragment thereof. In many instances, a polynucleotide encodes a polypeptide (or protein) or a domain or a fragment thereof. Additionally, a polynucleotide can comprise a promoter, an intron, an enhancer region, a polyadenylation site, a translation initiation site, 5' or 3' untranslated regions, a reporter gene, a selectable marker, a scorable marker, or the like. A polynucleotide can be single-stranded or double-stranded DNA or RNA.
[0110] A polynucleotide optionally comprises modified bases or a modified backbone. A polynucleotide can be, for example, genomic DNA or RNA, a transcript (such as an mRNA), a cDNA, a PCR product, a cloned DNA, a synthetic DNA or RNA, or the like. A polynucleotide can be combined with carbohydrate(s), lipid(s), protein(s), or other materials to perform a particular activity such as transformation or form a composition such as a peptide nucleic acid (PNA). A polynucleotide can comprise a sequence in either sense or antisense orientations. "Oligonucleotide" is substantially equivalent to the terms amplimer, primer, oligomer, element, target, and probe and is preferably single-stranded.
[0111] As used herein a "recombinant polynucleotide" or "recombinant DNA" is a polynucleotide that is not in its native state, for example, a polynucleotide comprises a series of nucleotides (represented as a nucleotide sequence) not found in nature, or a polynucleotide is in a context other than that in which it is naturally found; for example, separated from polynucleotides with which it typically is in proximity in nature, or adjacent (or contiguous with) polynucleotides with which it typically is not in proximity. The "recombinant polynucleotide" or "recombinant DNA" refers to polynucleotide or DNA which has been genetically engineered and constructed outside of a cell including DNA containing naturally occurring DNA or cDNA or synthetic DNA. For example, the polynucleotide at issue can be cloned into a vector, or otherwise recombined with one or more additional nucleic acids.
[0112] As used herein a "polypeptide" comprises a plurality of consecutive polymerized amino acid residues for example, at least about 15 consecutive polymerized amino acid residues. In many instances, a polypeptide comprises a series of polymerized amino acid residues that is a transcriptional regulator or a domain or portion or fragment thereof. Additionally, the polypeptide can comprise: (i) a localization domain; (ii) an activation domain; (iii) a repression domain; (iv) an oligomerization domain; (v) a protein-protein interaction domain; (vi) a DNA-binding domain; or the like. The polypeptide optionally comprises modified amino acid residues, naturally occurring amino acid residues not encoded by a codon, non-naturally occurring amino acid residues.
[0113] As used herein "protein" refers to a series of amino acids, oligopeptide, peptide, polypeptide or portions thereof whether naturally occurring or synthetic.
[0114] As used herein a "recombinant polypeptide" is a polypeptide produced by translation of a recombinant polynucleotide.
[0115] A "synthetic polypeptide" is a polypeptide created by consecutive polymerization of isolated amino acid residues using methods known in the art.
[0116] An "isolated polypeptide", whether a naturally occurring or a recombinant polypeptide, is more enriched in (or out of) a cell than the polypeptide in its natural state in a wild-type cell, for example, more than about 5% enriched, more than about 10% enriched, or more than about 20%, or more than about 50%, or more, enriched, for example, alternatively denoted: 105%, 110%, 120%, 150% or more, enriched relative to wild type standardized at 100%. Such enrichment is not the result of a natural response of a wild-type plant. Alternatively, or additionally, the isolated polypeptide is separated from other cellular components, with which it is typically associated, for example, by any of the various protein purification methods.
[0117] As used herein, a "functional fragment" refers to a portion of a polypeptide provided herein which retains full or partial molecular, physiological or biochemical function of the full length polypeptide. A functional fragment often contains the domain(s), such as Pfam domains (see below), identified in the polypeptide provided in the sequence listing.
[0118] A "recombinant DNA construct" as used in the present disclosure comprises at least one expression cassette having a promoter operable in plant cells and a polynucleotide of the present disclosure. DNA constructs can be used as a means of delivering recombinant DNA constructs to a plant cell in order to effect stable integration of the recombinant molecule into the plant cell genome. In one embodiment, the polynucleotide can encode a protein or variant of a protein or fragment of a protein that is functionally defined to maintain activity in transgenic host cells including plant cells, plant parts, explants and whole plants. In another embodiment, the polynucleotide can encode a non-coding RNA that interferes with the functioning of endogenous classes of small RNAs that regulate expression, including but not limited to taRNAs, siRNAs and miRNAs. Recombinant DNA constructs are assembled using methods known to persons of ordinary skill in the art and typically comprise a promoter operably linked to DNA, the expression of which provides the enhanced agronomic trait.
[0119] Other construct components can include additional regulatory elements, such as 5' leaders and introns for enhancing transcription, 3' untranslated regions (such as polyadenylation signals and sites), and DNA for transit or targeting or signal peptides.
[0120] Percent identity describes the extent to which polynucleotides or protein segments are invariant in an alignment of sequences, for example nucleotide sequences or amino acid sequences. An alignment of sequences is created by manually aligning two sequences, for example, a stated sequence, as provided herein, as a reference, and another sequence, to produce the highest number of matching elements, for example, individual nucleotides or amino acids, while allowing for the introduction of gaps into either sequence. An "identity fraction" for a sequence aligned with a reference sequence is the number of matching elements, divided by the full length of the reference sequence, not including gaps introduced by the alignment process into the reference sequence. "Percent identity" ("% identity") as used herein is the identity fraction times 100.
[0121] As used herein, a "homolog" or "homologues" means a protein in a group of proteins that perform the same biological function, for example, proteins that belong to the same Pfam protein family and that provide a common enhanced trait in transgenic plants of this disclosure. Homologs are expressed by homologous genes. With reference to homologous genes, homologs include orthologs, for example, genes expressed in different species that evolved from common ancestral genes by speciation and encode proteins retain the same function, but do not include paralogs, i.e., genes that are related by duplication but have evolved to encode proteins with different functions. Homologous genes include naturally occurring alleles and artificially-created variants.
[0122] Degeneracy of the genetic code provides the possibility to substitute at least one base of the protein encoding sequence of a gene with a different base without causing the amino acid sequence of the polypeptide produced from the gene to be changed. When optimally aligned, homolog proteins, or their corresponding nucleotide sequences, have typically at least about 60% identity, in some instances at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 92%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or even at least about 99.5% identity over the full length of a protein or its corresponding nucleotide sequence identified as being associated with imparting an enhanced trait or altered phenotype when expressed in plant cells. In one aspect of the disclosure homolog proteins have at least about 80%, at least about 85%, at least about 90%, at least about 92%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 99.5% identity to a consensus amino acid sequence of proteins and homologs that can be built from sequences disclosed herein.
[0123] Homologs are inferred from sequence similarity, by comparison of protein sequences, for example, manually or by use of a computer-based tool using known sequence comparison algorithms such as BLAST and FASTA. A sequence search and local alignment program, for example, BLAST, can be used to search query protein sequences of a base organism against a database of protein sequences of various organisms, to find similar sequences, and the summary Expectation value (E-value) can be used to measure the level of sequence similarity. Because a protein hit with the lowest E-value for a particular organism may not necessarily be an ortholog or be the only ortholog, a reciprocal query is used to filter hit sequences with significant E-values for ortholog identification. The reciprocal query entails search of the significant hits against a database of protein sequences of the base organism. A hit can be identified as an ortholog, when the reciprocal query's best hit is the query protein itself or a paralog of the query protein. With the reciprocal query process orthologs are further differentiated from paralogs among all the homologs, which allows for the inference of functional equivalence of genes. A further aspect of the homologs encoded by DNA useful in the transgenic plants of the invention are those proteins that differ from a disclosed protein as the result of deletion or insertion of one or more amino acids in a native sequence.
[0124] Other functional homolog proteins differ in one or more amino acids from those of a trait-improving protein disclosed herein as the result of one or more of known conservative amino acid substitutions, for example, valine is a conservative substitute for alanine and threonine is a conservative substitute for serine. Conservative substitutions for an amino acid within the native sequence can be selected from other members of a class to which the naturally occurring amino acid belongs. Representative amino acids within these various classes include, but are not limited to: (1) acidic (negatively charged) amino acids such as aspartic acid and glutamic acid; (2) basic (positively charged) amino acids such as arginine, histidine, and lysine; (3) neutral polar amino acids such as glycine, serine, threonine, cysteine, tyrosine, asparagine, and glutamine; and (4) neutral nonpolar (hydrophobic) amino acids such as alanine, leucine, isoleucine, valine, proline, phenylalanine, tryptophan, and methionine. Conserved substitutes for an amino acid within a native protein or polypeptide can be selected from other members of the group to which the naturally occurring amino acid belongs. For example, a group of amino acids having aliphatic side chains is glycine, alanine, valine, leucine, and isoleucine; a group of amino acids having aliphatic-hydroxyl side chains is serine and threonine; a group of amino acids having amide-containing side chains is asparagine and glutamine; a group of amino acids having aromatic side chains is phenylalanine, tyrosine, and tryptophan; a group of amino acids having basic side chains is lysine, arginine, and histidine; and a group of amino acids having sulfur-containing side 30 chains is cysteine and methionine. Naturally conservative amino acids substitution groups are: valine-leucine, valine-isoleucine, phenylalanine-tyrosine, lysine-arginine, alaninevaline, aspartic acid-glutamic acid, and asparagine-glutamine. A further aspect of the disclosure includes proteins that differ in one or more amino acids from those of a described protein sequence as the result of deletion or insertion of one or more amino acids in a native sequence.
[0125] In general, the term "variant" refers to molecules with some differences, generated synthetically or naturally, in their nucleotide or amino acid sequences as compared to a reference (native) polynucleotides or polypeptides, respectively. These differences include substitutions, insertions, deletions or any desired combinations of such changes in a native polynucleotide or amino acid sequence.
[0126] With regard to polynucleotide variants, differences between presently disclosed polynucleotides and polynucleotide variants are limited so that the nucleotide sequences of the former and the latter are similar overall and, in many regions, identical. Due to the degeneracy of the genetic code, differences between the former and the latter nucleotide sequences may be silent (for example, the amino acids encoded by the polynucleotide are the same, and the variant polynucleotide sequence encodes the same amino acid sequence as the presently disclosed polynucleotide). Variant nucleotide sequences can encode different amino acid sequences, in which case such nucleotide differences will result in amino acid substitutions, additions, deletions, insertions, truncations or fusions with respect to the similarly disclosed polynucleotide sequences. These variations can result in polynucleotide variants encoding polypeptides that share at least one functional characteristic. The degeneracy of the genetic code also dictates that many different variant polynucleotides can encode identical and/or substantially similar polypeptides.
[0127] As used herein "gene" or "gene sequence" refers to the partial or complete coding sequence of a gene, its complement, and its 5' and/or 3' untranslated regions (UTRs) and their complements. A gene is also a functional unit of inheritance, and in physical terms is a particular segment or sequence of nucleotides along a molecule of DNA (or RNA, in the case of RNA viruses) involved in producing a polypeptide chain. The latter can be subjected to subsequent processing such as chemical modification or folding to obtain a functional protein or polypeptide. By way of example, a transcriptional regulator gene encodes a transcriptional regulator polypeptide, which can be functional or require processing to function as an initiator of transcription.
[0128] As used herein, the term "promoter" refers generally to a DNA molecule that is involved in recognition and binding of RNA polymerase II and other proteins (trans-acting transcription factors) to initiate transcription. A promoter can be initially isolated from the 5' untranslated region (5' UTR) of a genomic copy of a gene. Alternately, promoters can be synthetically produced or manipulated DNA molecules. Promoters can also be chimeric, that is a promoter produced through the fusion of two or more heterologous DNA molecules. Plant promoters include promoter DNA obtained from plants, plant viruses, fungi and bacteria such as Agrobacterium and Bradyrhizobium bacteria.
[0129] Promoters which initiate transcription in all or most tissues of the plant are referred to as "constitutive" promoters. Promoters which initiate transcription during certain periods or stages of development are referred to as "developmental" promoters. Promoters whose expression is enhanced in certain tissues of the plant relative to other plant tissues are referred to as "tissue enhanced" or "tissue preferred" promoters. Promoters which express within a specific tissue of the plant, with little or no expression in other plant tissues are referred to as "tissue specific" promoters. A promoter that expresses in a certain cell type of the plant, for example a microspore mother cell, is referred to as a "cell type specific" promoter. An "inducible" promoter is a promoter in which transcription is initiated in response to an environmental stimulus such as cold, drought or light; or other stimuli such as wounding or chemical application. Many physiological and biochemical processes in plants exhibit endogenous rhythms with a period of about 24 hours. A "diurnal promoter" is a promoter which exhibits altered expression profiles under the control of a circadian oscillator. Diurnal regulation is subject to environmental inputs such as light and temperature and coordination by the circadian clock.
[0130] Sufficient expression in plant seed tissues is desired to affect improvements in seed composition. Exemplary promoters for use for seed composition modification include promoters from seed genes such as napin as disclosed in U.S. Pat. No. 5,420,034, maize L3 oleosin as disclosed in U.S. Pat. No. 6,433,252, zein Z27 as disclosed by Russell et al. (1997) Transgenic Res. 6(2):157-166, globulin 1 as disclosed by Belanger et al (1991) Genetics 129:863-872, glutelin 1 as disclosed by Russell (1997) supra, and peroxiredoxin antioxidant (Perl) as disclosed by Stacy et al. (1996) Plant Mol Biol. 31(6):1205-1216.
[0131] As used herein, the term "leader" refers to a DNA molecule isolated from the untranslated 5' region (5' UTR) of a genomic copy of a gene and is defined generally as a nucleotide segment between the transcription start site (TSS) and the protein coding sequence start site. Alternately, leaders can be synthetically produced or manipulated DNA elements. A leader can be used as a 5' regulatory element for modulating expression of an operably linked transcribable polynucleotide molecule. As used herein, the term "intron" refers to a DNA molecule that can be isolated or identified from the genomic copy of a gene and can be defined generally as a region spliced out during mRNA processing prior to translation. Alternately, an intron can be a synthetically produced or manipulated DNA element. An intron can contain enhancer elements that effect the transcription of operably linked genes. An intron can be used as a regulatory element for modulating expression of an operably linked transcribable polynucleotide molecule. A DNA construct can comprise an intron, and the intron may or may not be with respect to the transcribable polynucleotide molecule.
[0132] As used herein, the term "enhancer" or "enhancer element" refers to a cis-acting transcriptional regulatory element, a.k.a. cis-element, which confers an aspect of the overall expression pattern, but is usually insufficient alone to drive transcription, of an operably linked polynucleotide. Unlike promoters, enhancer elements do not usually include a transcription start site (TSS) or TATA box or equivalent sequence. A promoter can naturally comprise one or more enhancer elements that affect the transcription of an operably linked polynucleotide. An isolated enhancer element can also be fused to a promoter to produce a chimeric promoter cis-element, which confers an aspect of the overall modulation of gene expression. A promoter or promoter fragment can comprise one or more enhancer elements that effect the transcription of operably linked genes. Many promoter enhancer elements are believed to bind DNA-binding proteins and/or affect DNA topology, producing local conformations that selectively allow or restrict access of RNA polymerase to the DNA template or that facilitate selective opening of the double helix at the site of transcriptional initiation. An enhancer element can function to bind transcription factors that regulate transcription. Some enhancer elements bind more than one transcription factor, and transcription factors can interact with different affinities with more than one enhancer domain.
[0133] Expression cassettes of this disclosure can include a "transit peptide" or "targeting peptide" or "signal peptide" molecule located either 5' or 3' to or within the gene(s). These terms generally refer to peptide molecules that when linked to a protein of interest directs the protein to a particular tissue, cell, subcellular location, or cell organelle. Examples include, but are not limited to, chloroplast transit peptides (CTPs), chloroplast targeting peptides, mitochondrial targeting peptides, nuclear targeting signals, nuclear exporting signals, vacuolar targeting peptides, and vacuolar sorting peptides. For description of the use of chloroplast transit peptides see U.S. Pat. Nos. 5,188,642 and 5,728,925. For description of the transit peptide region of an Arabidopsis EPSPS gene in the present disclosure, see Klee, H. J. Et al (MGG (1987) 210:437-442. Expression cassettes of this disclosure can also include an intron or introns. Expression cassettes of this disclosure can contain a DNA near the 3' end of the cassette that acts as a signal to terminate transcription from a heterologous nucleic acid and that directs polyadenylation of the resultant mRNA. These are commonly referred to as "3'-untranslated regions" or "3'-non-coding sequences" or "3'-UTRs". The "3' non-translated sequences" means DNA sequences located downstream of a structural nucleotide sequence and include sequences encoding polyadenylation and other regulatory signals capable of affecting mRNA processing or gene expression. The polyadenylation signal functions in plants to cause the addition of polyadenylate nucleotides to the 3' end of the mRNA precursor. The polyadenylation signal can be derived from a natural gene, from a variety of plant genes, or from T-DNA. An example of a polyadenylation sequence is the nopaline synthase 3' sequence (nos 3'; Fraley et al., Proc. Natl. Acad. Sci. USA 80: 4803-4807, 1983). The use of different 3' non-translated sequences is exemplified by Ingelbrecht et al., Plant Cell 1:671-680, 1989.
[0134] Expression cassettes of this disclosure can also contain one or more genes that encode selectable markers and confer resistance to a selective agent such as an antibiotic or an herbicide. A number of selectable marker genes are known in the art and can be used in the present disclosure: selectable marker genes conferring tolerance to antibiotics like kanamycin and paromomycin (nptII), hygromycin B (aph IV), spectinomycin (aadA), U.S. Patent Publication 2009/0138985A1 and gentamycin (aac3 and aacC4) or tolerance to herbicides like glyphosate (for example, 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), U.S. Pat. Nos. 5,627,061; 5,633,435; 6,040,497; 5,094,945), sulfonyl herbicides (for example, acetohydroxyacid synthase or acetolactate synthase conferring tolerance to acetolactate synthase inhibitors such as sulfonylurea, imidazolinone, triazolopyrimidine, pyrimidyloxybenzoates and phthalide (U.S. Pat. Nos. 6,225,105; 5,767,366; 4,761,373; 5,633,437; 6,613,963; 5,013,659; 5,141,870; 5,378,824; 5,605,011)), bialaphos or phosphinothricin or derivatives (e, g., phosphinothricin acetyltransferase (bar) tolerance to phosphinothricin or glufosinate (U.S. Pat. Nos. 5,646,024; 5,561,236; 5,276,268; 5,637,489; 5,273,894); dicamba (dicamba monooxygenase, Patent Application Publications US2003/0115626A1), or sethoxydim (modified acetyl-coenzyme A carboxylase for conferring tolerance to cyclohexanedione (sethoxydim)), and aryloxyphenoxypropionate (haloxyfop, U.S. Pat. No. 6,414,222).
[0135] Transformation vectors of this disclosure can contain one or more "expression cassettes", each comprising a native or non-native plant promoter operably linked to a polynucleotide sequence of interest, which is operably linked to a 3' UTR termination signal, for expression in an appropriate host cell. It also typically comprises sequences required for proper translation of the polynucleotide or transgene. As used herein, the term "transgene" refers to a polynucleotide molecule artificially incorporated into a host cell's genome. Such a transgene can be heterologous to the host cell. The term "transgenic plant" refers to a plant comprising such a transgene. The coding region usually codes for a protein of interest but can also code for a functional RNA of interest, for example an antisense RNA, a nontranslated RNA, in the sense or antisense direction, a miRNA, a noncoding RNA, or a synthetic RNA used in either suppression or over expression of target gene sequences. The expression cassette comprising the nucleotide sequence of interest can be chimeric, meaning that at least one of its components is heterologous with respect to at least one of its other components. As used herein the term "chimeric" refers to a DNA molecule that is created from two or more genetically diverse sources, for example a first molecule from one gene or organism and a second molecule from another gene or organism.
[0136] Recombinant DNA constructs in this disclosure generally include a 3' element that typically contains a polyadenylation signal and site. Known 3' elements include those from Agrobacterium tumefaciens genes such as nos 3', tml 3', tmr 3', tms 3', ocs 3', tr7 3', for example disclosed in U.S. Pat. No. 6,090,627; 3' elements from plant genes such as wheat (Triticum aesevitum) heat shock protein 17 (Hsp17 3'), a wheat ubiquitin gene, a wheat fructose-1,6-biphosphatase gene, a rice glutelin gene, a rice lactate dehydrogenase gene and a rice beta-tubulin gene, all of which are disclosed in US Patent Application Publication 2002/0192813 A1; and the pea (Pisum sativum) ribulose biphosphate carboxylase gene (rbs 3'), and 3' elements from the genes within the host plant.
[0137] As used herein "operably linked" means the association of two or more DNA fragments in a recombinant DNA construct so that the function of one, for example, protein-encoding DNA, is controlled by the other, for example, a promoter.
[0138] Transgenic plants can comprise a stack of one or more polynucleotides disclosed herein resulting in the production of multiple polypeptide sequences. Transgenic plants comprising stacks of polynucleotides can be obtained by either or both of traditional breeding methods or through genetic engineering methods. These methods include, but are not limited to, crossing individual transgenic lines each comprising a polynucleotide of interest, transforming a transgenic plant comprising a first gene disclosed herein with a second gene, and co-transformation of genes into a single plant cell. Co-transformation of genes can be carried out using single transformation vectors comprising multiple genes or genes carried separately on multiple vectors.
[0139] Transgenic plants comprising or derived from plant cells of this disclosure transformed with recombinant DNA can be further enhanced with stacked traits, for example, a crop plant having an enhanced trait resulting from expression of DNA disclosed herein in combination with herbicide and/or pest resistance traits. For example, genes of the current disclosure can be stacked with other traits of agronomic interest, such as a trait providing herbicide resistance, or insect resistance, such as using a gene from Bacillus thuringensis to provide resistance against lepidopteran, coliopteran, homopteran, hemiopteran, and other insects, or improved quality traits such as improved nutritional value. Herbicides for which transgenic plant tolerance has been demonstrated and the method of the present disclosure can be applied include, but are not limited to, glyphosate, dicamba, glufosinate, sulfonylurea, bromoxynil and norflurazon herbicides. Polynucleotide molecules encoding proteins involved in herbicide tolerance known in the art and include, but are not limited to, a polynucleotide molecule encoding 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) disclosed in U.S. Pat. Nos. 5,094,945; 5,627,061; 5,633,435 and 6,040,497 for imparting glyphosate tolerance; polynucleotide molecules encoding a glyphosate oxidoreductase (GOX) disclosed in U.S. Pat. No. 5,463,175 and a glyphosate-N-acetyl transferase (GAT) disclosed in US Patent Application Publication 2003/0083480 A1 also for imparting glyphosate tolerance; dicamba monooxygenase disclosed in US Patent Application Publication 2003/0135879 A1 for imparting dicamba tolerance; a polynucleotide molecule encoding bromoxynil nitrilase (Bxn) disclosed in U.S. Pat. No. 4,810,648 for imparting bromoxynil tolerance; a polynucleotide molecule encoding phytoene desaturase (crtI) described in Misawa et al, (1993) Plant J. 4:833-840 and in Misawa et al, (1994) Plant J. 6:481-489 for norflurazon tolerance; a polynucleotide molecule encoding acetohydroxyacid synthase (AHAS, aka ALS) described in Sathasiivan et al. (1990) Nucl. Acids Res. 18:2188-2193 for imparting tolerance to sulfonylurea herbicides; polynucleotide molecules known as bar genes disclosed in DeBlock, et al. (1987) EMBO J. 6:2513-2519 for imparting glufosinate and bialaphos tolerance; polynucleotide molecules disclosed in US Patent Application Publication 2003/010609 A1 for imparting N-amino methyl phosphonic acid tolerance; polynucleotide molecules disclosed in U.S. Pat. No. 6,107,549 for imparting pyridine herbicide resistance; molecules and methods for imparting tolerance to multiple herbicides such as glyphosate, atrazine, ALS inhibitors, isoxoflutole and glufosinate herbicides are disclosed in U.S. Pat. No. 6,376,754 and US Patent Application Publication 2002/0112260. Molecules and methods for imparting insect/nematode/virus resistance are disclosed in U.S. Pat. Nos. 5,250,515; 5,880,275; 6,506,599; 5,986,175 and US Patent Application Publication 2003/0150017 A1.
Plant Cell Transformation Methods
[0140] Numerous methods for transforming chromosomes in a plant cell with recombinant DNA are known in the art and are used in methods of producing a transgenic plant cell and plant. Two effective methods for such transformation are Agrobacterium-mediated transformation and microprojectile bombardment-mediated transformation. Microprojectile bombardment methods are illustrated in U.S. Pat. No. 5,015,580 (soybean); U.S. Pat. No. 5,550,318 (corn); U.S. Pat. No. 5,538,880 (corn); U.S. Pat. No. 5,914,451 (soybean); U.S. Pat. No. 6,160,208 (corn); U.S. Pat. No. 6,399,861 (corn); U.S. Pat. No. 6,153,812 (wheat) and U.S. Pat. No. 6,365,807 (rice). Agrobacterium-mediated transformation methods are described in U.S. Pat. No. 5,159,135 (cotton); U.S. Pat. No. 5,824,877 (soybean); U.S. Pat. No. 5,463,174 (canola); U.S. Pat. No. 5,591,616 (corn); U.S. Pat. No. 5,846,797 (cotton); U.S. Pat. No. 8,044,260 (cotton); U.S. Pat. No. 6,384,301 (soybean), U.S. Pat. No. 7,026,528 (wheat) and U.S. Pat. No. 6,329,571 (rice), US Patent Application Publication 2004/0087030 A1 (cotton), and US Patent Application Publication 2001/0042257 A1 (sugar beet), all of which are incorporated herein by reference in their entirety. Transformation of plant material is practiced in tissue culture on nutrient media, for example a mixture of nutrients that allow cells to grow in vitro. Recipient cell targets include, but are not limited to, meristem cells, shoot tips, hypocotyls, calli, immature or mature embryos, and gametic cells such as microspores, pollen, sperm and egg cells. Callus can be initiated from tissue sources including, but not limited to, immature or mature embryos, hypocotyls, seedling apical meristems, microspores and the like. Cells containing a transgenic nucleus are grown into transgenic plants.
[0141] In addition to direct transformation of a plant material with a recombinant DNA construct, a transgenic plant can be prepared by crossing a first plant comprising a recombinant DNA with a second plant lacking the recombinant DNA. For example, recombinant DNA can be introduced into a first plant line that is amenable to transformation, which can be crossed with a second plant line to introgress the recombinant DNA into the second plant line. A transgenic plant with recombinant DNA providing an enhanced trait, for example, enhanced yield, can be crossed with a transgenic plant line having other recombinant DNA that confers another trait, for example herbicide resistance or pest resistance, to produce progeny plants having recombinant DNA that confers both traits. Typically, in such breeding for combining traits the transgenic plant donating the additional trait is a male line and the transgenic plant carrying the base traits is the female line. The progeny of this cross will segregate such that some of the plants will carry the DNA for both parental traits and some will carry DNA for one parental trait; such plants can be identified by markers associated with parental recombinant DNA, for example, marker identification by analysis for recombinant DNA or, in the case where a selectable marker is linked to the recombinant, by application of the selecting agent such as a herbicide for use with a herbicide tolerance marker, or by selection for the enhanced trait. Progeny plants carrying DNA for both parental traits can be crossed back into the female parent line multiple times, for example usually 6 to 8 generations, to produce a progeny plant with substantially the same genotype as the original transgenic parental line but for the recombinant DNA of the other transgenic parental line.
[0142] For transformation, DNA is typically introduced into only a small percentage of target plant cells in any one transformation experiment. Marker genes are used to provide an efficient system for identification of those cells that are stably transformed by receiving and integrating a recombinant DNA construct into their genomes. Preferred marker genes provide selective markers which confer resistance to a selective agent, such as an antibiotic or an herbicide. Any of the herbicides to which plants of this disclosure can be resistant is an agent for selective markers. Potentially transformed cells are exposed to the selective agent. In the population of surviving cells are those cells where, generally, the resistance-conferring gene is integrated and expressed at sufficient levels to permit cell survival. Cells can be tested further to confirm stable integration of the exogenous DNA. Commonly used selective marker genes include those conferring resistance to antibiotics such as kanamycin and paromomycin (npII), hygromycin B (aph IV), spectinomycin (aadA) and gentamycin (aac3 and aacC4) or resistance to herbicides such as glufosinate (bar or pat), dicamba (DMO) and glyphosate (aroA or EPSPS). Examples of such selectable markers are illustrated in U.S. Pat. Nos. 5,550,318; 5,633,435; 5,780,708; 6,118,047 and 8,030,544. Markers which provide an ability to visually screen transformants can also be employed, for example, a gene expressing a colored or fluorescent protein such as a luciferase or green fluorescent protein (GFP) or a gene expressing a beta-glucuronidase or uidA gene (GUS) for which various chromogenic substrates are known.
[0143] Plant cells that survive exposure to a selective agent, or plant cells that have been scored positive in a screening assay, may be cultured in vitro to regenerate plantlets. Developing plantlets regenerated from transformed plant cells can be transferred to plant growth mix, and hardened off, for example, in an environmentally controlled chamber at about 85% relative humidity, 600 ppm CO.sub.2, and 25-250 microeinsteins m.sup.-2 s.sup.-1 of light, prior to transfer to a greenhouse or growth chamber for maturation. Plants are regenerated from about 6 weeks to 10 months after a transformant is identified, depending on the initial tissue, and plant species. Plants can be pollinated using conventional plant breeding methods known to those of skill in the art to produce seeds, for example self-pollination is commonly used with transgenic corn. The regenerated transformed plant or its progeny seed or plants can be tested for expression of the recombinant DNA and selected for the presence of an enhanced agronomic trait.
Transgenic Plants and Seeds
[0144] Transgenic plants derived from transgenic plant cells having a transgenic nucleus of this disclosure are grown to generate transgenic plants having an enhanced trait as compared to a control plant, and produce transgenic seed and haploid pollen of this disclosure. Such plants with enhanced traits are identified by selection of transformed plants or progeny seed for the enhanced trait. For efficiency a selection method is designed to evaluate multiple transgenic plants (events) comprising the recombinant DNA, for example multiple plants from 2 to 20 or more transgenic events. Transgenic plants grown from transgenic seeds provided herein demonstrate improved agronomic traits that contribute to increased yield or other traits that provide increased plant value, including, for example, improved seed quality. Of particular interest are plants having increased water use efficiency or drought tolerance, enhanced high temperature or cold tolerance, increased yield, and increased nitrogen use efficiency.
[0145] Table 1 provides a list of sequences of protein-encoding genes as recombinant DNA for production of transgenic plants with enhanced traits. The elements of Table 1 are described by reference to:
[0146] "NUC SEQ ID NO." which identifies a DNA sequence.
[0147] "PEP SEQ ID NO." which identifies an amino acid sequence.
[0148] "Gene ID" which refers to an arbitrary identifier.
[0149] "Gene Name and Description" which is a common name and functional description of the gene.
TABLE-US-00001 TABLE 1 Sequences for Protein-Coding Genes NUC PEP SEQ SEQ ID ID NO. NO. Gene ID Gene Name and Description 1 28 TRDX4-01 Arabidopsis mitochondrial import receptor subunit TOMS homolog (TOM5) 2 29 TRDX4-02 Arabidopsis K+ independent Asparaginase 3 30 TRDX4-03 Arabidopsis plasma membrane (PM)- localized cyclic nucleotide gated channels (CNGCs) 4 31 TRDX4-04 Arabidopsis receptor like kinase 5 32 TRDX4-05 Rice gibberellin receptor gene GID1 6 33 TRDX4-06 Corn plastidial phosphoenolpyruvate (PEP) phosphate translocator (PPT) 7 34 TRDX4-07 Arabidopsis sulfolipid biosynthesis protein SQD1 8 35 TRDX4-08 Arabidopsis cytochrome P450 family protein 9 36 TRDX4-09 Pseudomonas syringae phosphoglycerate kinase 10 37 TRDX4-10 Corn phospholipase A (PLA1) 11 38 TRDX4-11 Arabidopsis plastidal glycolate/ glycerate translocator 1 (PLGG1) 12 39 TRDX4-12 Corn coiled coil domain protein 13 40 TRDX4-13 Corn iron-phytosiderophore transporter protein yellow stripe 1 (YS1) 14 41 TRDX4-14 Arabidopsis ACT domain-containing protein 3 (ACR3) 15 42 TRDX4-15 E. coli arginine-insensitive acetylglutarnate kinase (NAGK) 16 43 TRDX4-16 Soybean NOS1 (mitochondrial constitutive NOS) 17 44 TRDX4-17 Corn thylakoid lumen protein CYP38 18 45 TRDX4-18 Arabidopsis glutaredoxin family protein 19 46 TRDX4-19 E. coli aminobutyrate aminotransferase 20 47 TRDX4-20 Synechocystis sp. gene of unknown function 21 48 TRDX4-21 Corn putative forever young oxidoreductase 22 49 TRDX4-22 Corn MSI-12 gene 23 50 TRDX4-23 Arabidopsis mitogen-activated protein kinase kinase kinase 19 (MAPKKK19) 24 51 TRDX4-24 Arabidopsis carbamoyl phosphate synthase EC 6.33.5 large subunit 25 52 TRDX4-25 Soybean gene improving Nitrogen Utilization Efficiency (NUE) 26 53 TRDX4-26 Arabidopsis casparian strip membrane protein 1 (CASP1) 27 54 TRDX4-27 E. coli codon redesigned asparagine synthetase A (AsnA) gene
[0150] Table 2 provides a list of sequences for suppression of target protein-coding genes, as recombinant DNA for production of transgenic plants with enhanced traits. The elements of Table 2 are described by reference to:
[0151] "Target NUC SEQ ID NO." which identifies a nucleotide coding sequence of the suppression target gene.
[0152] "Target PEP SEQ ID NO." which identifies an amino acid sequence of the suppression target gene.
[0153] "Target Gene ID" which is an arbitrary identifier of the suppression target gene.
[0154] "Engineered miRNA precursor SEQ ID NO." which identifies a nucleotide sequence of the miRNA construct.
[0155] "miRNA recognition site SEQ ID NO." which identifies a nucleotide sequence of the miRNA recognition site.
[0156] "Target Gene Name and Description" which is a common name and functional description of the suppression target gene.
TABLE-US-00002 TABLE 2 Sequences for Gene Suppression Engi- neered miRNA Target Target miRNA recog- NUC PEP pre- nition SEQ SEQ Target cursor site Target ID ID Gene SEQ SEQ Gene Name NO. NO. ID ID NO. ID NO. and Description 55 61 TRDX4-1T 67 73 corn homolog of NOX1 gene, Plastidial phosphoenol- pyruvate (PEP) phosphate translocator PPT) 56 62 TRDX4-3T 68 74 soybean SOUL gene 57 63 TRDX4-4T 69 75 soybean Elongated Hypocotyl 5 (Hy5) 58 64 TRDX4-5T 70 76 corn Proliferating cell nuclear antigen 2 (PCNA2) 59 65 TRDX4-6T 71 77 corn putative dolichyl-di- phosphooligo- saccharide protein 60 66 TRDX4-7T 72 78 corn Peroxi- somal_fatty_acid_beta- oxidation
Selection Methods for Transgenic Plants with Enhanced Traits
[0157] Within a population of transgenic plants each regenerated from a plant cell with recombinant DNA many plants that survive to fertile transgenic plants that produce seeds and progeny plants will not exhibit an enhanced agronomic trait. Selection from the population is necessary to identify one or more transgenic plants with an enhanced trait. Transgenic plants having enhanced traits are selected from populations of plants regenerated or derived from plant cells transformed as described herein by evaluating the plants in a variety of assays to detect an enhanced trait, for example, increased water use efficiency or drought tolerance, enhanced high temperature or cold tolerance, increased yield, increased nitrogen use efficiency, enhanced seed composition such as enhanced seed protein and enhanced seed oil. These assays can take many forms including, but not limited to, direct screening for the trait in a greenhouse or field trial or by screening for a surrogate trait. Such analyses can be directed to detecting changes in the chemical composition, biomass, physiological property, or morphology of the plant. Changes in chemical compositions such as nutritional composition of grain can be detected by analysis of the seed composition and content of protein, free amino acids, oil, free fatty acids, starch or tocopherols. Changes in chemical compositions can also be detected by analysis of contents in leaves, such as chlorophyll or carotenoid contents. Changes in biomass characteristics can be evaluated on greenhouse or field grown plants and can include plant height, stem diameter, root and shoot dry weights, canopy size; and, for corn plants, ear length and diameter. Changes in physiological properties can be identified by evaluating responses to stress conditions, for example assays using imposed stress conditions such as water deficit, nitrogen deficiency, cold growing conditions, pathogen or insect attack or light deficiency, or increased plant density. Changes in morphology can be measured by visual observation of tendency of a transformed plant to appear to be a normal plant as compared to changes toward bushy, taller, thicker, narrower leaves, striped leaves, knotted trait, chlorosis, albino, anthocyanin production, or altered tassels, ears or roots. Other selection properties include days to pollen shed, days to silking, leaf extension rate, chlorophyll content, leaf temperature, stand, seedling vigor, internode length, plant height, leaf number, leaf area, tillering, brace roots, stay green or delayed senescence, stalk lodging, root lodging, plant health, bareness/prolificacy, green snap, and pest resistance. In addition, phenotypic characteristics of harvested grain can be evaluated, including number of kernels per row on the ear, number of rows of kernels on the ear, kernel abortion, kernel weight, kernel size, kernel density and physical grain quality.
[0158] Assays for screening for a desired trait are readily designed by those practicing in the art. The following illustrates screening assays for corn traits using hybrid corn plants. The assays can be adapted for screening other plants such as canola, wheat, cotton and soybean either as hybrids or inbreds.
[0159] Transgenic corn plants having increased nitrogen use efficiency can be identified by screening transgenic plants in the field under the same and sufficient amount of nitrogen supply as compared to control plants, where such plants provide higher yield as compared to control plants. Transgenic corn plants having increased nitrogen use efficiency can also be identified by screening transgenic plants in the field under reduced amount of nitrogen supply as compared to control plants, where such plants provide the same or similar yield as compared to control plants.
[0160] Transgenic corn plants having increased yield are identified by screening using progenies of the transgenic plants over multiple locations for several years with plants grown under optimal production management practices and maximum weed and pest control or standard agronomic practices (SAP). Selection methods can be applied in multiple and diverse geographic locations, for example up to 16 or more locations, over one or more planting seasons, for example at least two planting seasons, to statistically distinguish yield improvement from natural environmental effects.
[0161] Transgenic corn plants having increased water use efficiency or drought tolerance are identified by screening plants in an assay where water is withheld for a period to induce stress followed by watering to revive the plants. For example, a selection process imposes 3 drought/re-water cycles on plants over a total period of 15 days after an initial stress free growth period of 11 days. Each cycle consists of 5 days, with no water being applied for the first four days and a water quenching on the 5th day of the cycle. The primary phenotypes analyzed by the selection method are the changes in plant growth rate as determined by height and biomass during a vegetative drought treatment.
[0162] Although the plant cells and methods of this disclosure can be applied to any plant cell, plant, seed or pollen, for example, any fruit, vegetable, grass, tree or ornamental plant, the various aspects of the disclosure are applied to corn, soybean, cotton, canola, rice, barley, oat, wheat, turf grass, alfalfa, sugar beet, sunflower, quinoa and sugar cane plants.
Example 1. Corn Transformation
[0163] This example illustrates transformation methods in producing a transgenic corn plant cell, seed, and plant having altered phenotypes as shown in Tables 4-6, or an enhanced trait, for example, increased water use efficiency, increased nitrogen use efficiency, and increased yield as shown in Tables 7 and 9.
[0164] For Agrobacterium-mediated transformation of corn embryo cells corn plants were grown in the greenhouse and ears were harvested when the embryos were 1.5 to 2.0 mm in length. Ears were surface-sterilized by spraying or soaking the ears in 80% ethanol, followed by air drying. Immature embryos were isolated from individual kernels on surface-sterilized ears. Shortly after excision, immature maize embryos were inoculated with overnight grown Agrobacterium cells, and incubated at room temperature with Agrobacterium for 5-20 minutes. Inoculated immature embryos were then co-cultured with Agrobacterium for 1 to 3 days at 23.degree. C. in the dark. Co-cultured embryos were transferred to selection media and cultured for approximately two weeks to allow embryogenic callus to develop. Embryogenic calli were transferred to culture medium containing glyphosate and subcultured at about two week intervals. Transformed plant cells were recovered 6 to 8 weeks after initiation of selection.
[0165] For Agrobacterium-mediated transformation of maize callus immature embryos are cultured for approximately 8-21 days after excision to allow callus to develop. Callus is then incubated for about 30 minutes at room temperature with the Agrobacterium suspension, followed by removal of the liquid by aspiration. The callus and Agrobacterium are co-cultured without selection for 3-6 days followed by selection on paromomycin for approximately 6 weeks, with biweekly transfers to fresh media. Paromomycin resistant calli are identified about 6-8 weeks after initiation of selection.
[0166] To regenerate transgenic corn plants individual transgenic calli resulting from transformation and selection were placed on media to initiate shoot and root development into plantlets. Plantlets were transferred to potting soil for initial growth in a growth chamber at 26.degree. C. followed by a mist bench before transplanting to 5 inch pots where plants were grown to maturity. The regenerated plants were self-fertilized and seeds were harvested for use in one or more methods to select seeds, seedlings or progeny second generation transgenic plants (R2 plants) or hybrids, for example, by selecting transgenic plants exhibiting an enhanced trait as compared to a control plant.
[0167] The above process can be repeated to produce multiple events of transgenic corn plants from cells that were transformed with recombinant DNA from the genes identified in Table 1 or with recombinant DNA from Table 2 that is transcribed into a non-coding miRNA. Progeny transgenic plants and seeds of the transformed plants were screened for the presence and single copy of the inserted gene, and for increased water use efficiency, increased yield, increased nitrogen use efficiency, and altered phenotypes as shown in Tables 4-6. From each group of multiple events of transgenic plants with a specific recombinant DNA from Table 1 or Table 2, the event(s) that showed increased yield, increased water use efficiency, increased nitrogen use efficiency, and altered phenotypes was (were) identified.
Example 2. Soybean Transformation
[0168] This example illustrates plant transformation in producing a transgenic soybean plant cell, seed, and plant having altered phenotypes, or an enhanced trait, for example, increased water use efficiency or drought tolerance and increased yield as shown in Tables 7 and 9.
[0169] For Agrobacterium mediated transformation, soybean seeds were imbibed overnight and the meristem explants excised. Soybean explants were mixed with induced Agrobacterium cells containing plasmid DNA with the gene of interest cassette and a plant selectable marker cassette no later than 14 hours from the time of initiation of seed imbibition, and wounded using sonication. Following wounding, explants were placed in co-culture for 2-5 days at which point they were transferred to selection media to allow selection and growth of transgenic shoots. Resistant shoots were harvested in approximately 6-8 weeks and placed into selective rooting media for 2-3 weeks. Shoots producing roots were transferred to the greenhouse and potted in soil. Shoots that remained healthy on selection, but did not produce roots were transferred to non-selective rooting media for an additional two weeks. Roots from any shoots that produced roots off selection were tested for expression of the plant selectable marker before they were transferred to the greenhouse and potted in soil.
[0170] The above process can be repeated to produce multiple events of transgenic soybean plants from cells that were transformed with recombinant DNA from the genes identified in Table 1 or recombinant DNA transcribed into a miRNA identified in Table 2. Progeny transgenic plants and seed of the transformed plant cells were screened for the presence and single copy of the inserted gene, and for increased water use efficiency and increased yield as shown in Tables 7 and 9.
Example 3. Identification of Altered Phenotypes in Automated Greenhouse
[0171] This example illustrates screening and identification of transgenic plants for altered phenotypes in an automated greenhouse (AGH). The apparatus and the methods for automated phenotypic screening of plants are disclosed in US Patent publication No. US20110135161 (filed on Nov. 10, 2010), which is incorporated by reference herein in its entirety.
Screening and Identification of Transgenic Corn Plants for Altered Phenotypes.
[0172] Corn plants were tested in 3 screens in AGH under different conditions including non-stress, nitrogen deficit and water deficit stress conditions. All screens began with a non-stress condition during day 0-5 germination phase, after which the plants were grown for 22 days under screen specific conditions as shown in Table 3.
TABLE-US-00003 TABLE 3 Description of the 3 AGH screens for corn plants Germination Screen specific phase phase Screen Description (5 days) (22 days) Non-stress well watered 55% VWC 55% VWC sufficient nitrogen water 8 mM nitrogen Water deficit limited watered 55% VWC 30% VWC sufficient nitrogen water 8 mM nitrogen Nitrogen deficit well watered 55% VWC 55% VWC low nitrogen water 2 mM nitrogen
[0173] Water deficit is defined as a specific Volumetric Water Content (VWC) that is lower than the VWC of non-stress plant. For example, a non-stressed plant might be maintained at 55% VWC and the VWC for a water-deficit assay might be defined around 30% VWC as shown in Table 3. Data were collected using visible light and hyperspectral imaging as well as direct measurement of pot weight and amount of water and nutrient applied to individual plants on a daily basis.
[0174] Nitrogen deficit is defined in part as a specific mM concentration of nitrogen that is lower than the nitrogen concentration of non-stress plants. For example, a non-stress plant might be maintained at 8 mM nitrogen while the nitrogen concentration applied in a nitrogen-deficit assay might be maintained at a concentration of 2 mM.
[0175] Up to ten parameters were measured for each screen. The visible light color imaging based measurements are: biomass, canopy area and plant height. Biomass (Bmass) is defined as estimated shoot fresh weight (g) of the plant obtained from images acquired from multiple angles of view. Canopy Area (Cnop) is defined as area of leaf as seen in top-down image (mm.sup.2). Plant Height (PlntH) refers to the distance from the top of the pot to the highest point of the plant derived from side image (mm). Anthocyanin score and area, chlorophyll score and concentration, and water content score are hyperspectral imaging based parameters. Anthocyanin Score (AntS) is an estimate of anthocyanin in the leaf canopy obtained from a top-down hyperspectral image. Anthocyanin Area (AntA) is an estimate of anthocyanin in the stem obtained from a side-view hyperspectral image. Chlorophyll Score (ClrpS) and Cholrophyll Concentration (ClrpC) are both measurements of chlorophyll in the leaf canopy obtained from a top-down hyperspectral image, where Chlorophyll Score measures in relative units and is done for soybean plants, and Chlorophyll Concentration measures in ppm units and is done for corn plants. Water Content Score (WtrCt) is a measurement of water in the leaf canopy obtained from a top-down hyperspectral image. Water Use Efficiency (WUE) is derived from the grams of plant biomass per liter of water added. Water Applied (WtrAp) is a direct measurement of water added to a pot (pot with no hole) during the course of an experiment.
[0176] These physiological screen runs were set up so that tested transgenic lines were compared to a control line. The collected data were analyzed against the control using % delta and certain p-value cutoff. Tables 4-6 are summaries of transgenic corn plants comprising the disclosed recombinant DNA constructs with altered phenotypes under non stress, nitrogen deficit, and water deficit conditions, respectively.
[0177] The test results are represented by three numbers: the first number before letter "p" denotes number of events with an increase in the tested parameter at p.ltoreq.0.1; the second number before letter "n" denotes number of events with an decrease in the tested parameter at p.ltoreq.0.1; the third number before letter "t" denotes total number of transgenic events tested for a given parameter in a specific screen. The increase or decrease is measured in comparison to non-transgenic control plants. A "-" means that it has not been tested. For example, 2p1n5t indicates that 5 transgenic plant events were screened, of which 2 events showed increase and 1 showed decrease of the measured parameter. Note that two constructs of gene TRDX4-19 were tested, and the results are listed as TRDX4-19 and TRDX4-19x.
TABLE-US-00004 TABLE 4 Summary of transgenic corn plants with altered phenotypes in AGH non-stress screens Gene_ID AntA AntS Bmass ClrpC ClrpS Cnop PlntH WUE WtrAp WtrCt TRDX4-01 0p0n5t 1p0n5t 0p2n5t 0p1n5t -- 0p3n5t 0p0n5t 0p1n5t 0p3n5t -- TRDX4-02 0p1n5t 0p1n5t 0p2n5t 0p2n5t -- 0p3n5t 0p2n5t 0p2n5t 0p3n5t -- TRDX4-03 -- 0p2n5t 0p1n5t 1p0n5t -- 0p2n5t 0p1n5t 0p2n5t 0p1n5t -- TRDX4-04 1p0n5t 0p0n5t 0p2n5t 0p0n5t -- 0p1n5t 0p3n5t 0p1n5t 0p1n5t -- TRDX4-05 1p0n5t 0p0n5t 0p0n5t 0p0n5t -- 0p0n5t 0p1n5t 0p0n5t 0p1n5t -- TRDX4-07 -- 0p0n5t 1p0n5t -- 1p0n5t 1p0n5t 0p1n5t 0p0n5t 3p0n5t 1p0n5t TRDX4-09 0p0n5t 0p0n5t 0p0n5t 0p0n5t -- 0p0n5t 1p0n5t 0p2n5t 1p0n5t -- TRDX4-11 0p1n5t 0p0n5t 0p1n5t 0p0n5t -- 0p0n5t 0p1n5t 0p1n5t 1p1n5t -- TRDX4-12 0p2n5t 0p2n5t 0p0n5t 1p0n5t -- 0p0n5t 0p0n5t 0p0n5t 2p0n5t -- TRDX4-13 1p0n5t 0p0n5t 0p0n5t 0p1n5t -- 0p1n5t 0p1n5t 0p0n5t 0p0n5t -- TRDX4-14 0p0n5t 1p0n5t 0p2n5t 1p0n5t -- 0p2n5t 0p4n5t 0p2n5t 0p2n5t -- TRDX4-16 0p0n5t 0p0n5t 0p0n5t 0p0n5t -- 0p1n5t 0p1n5t 0p0n5t 0p0n5t -- TRDX4-17 0p1n5t 0p0n5t 1p0n5t 1p0n5t -- 2p0n5t 0p0n5t 0p0n5t 4p0n5t -- TRDX4-18 0p2n5t 0p0n5t 2p1n5t 0p0n5t -- 1p1n5t 0p1n5t 1p1n5t 3p1n5t -- TRDX4-19 0p1n4t 0p0n4t 0p1n4t 0p0n4t -- 0p1n4t 0p1n4t 0p1n4t 0p1n4t -- TRDX4-19x 1p0n5t 1p0n5t 0p0n5t 0p0n5t -- 0p3n5t 0p1n5t 0p0n5t 0p1n5t -- TRDX4-1T 0p0n5t 0p0n5t 0p1n5t 3p0n5t -- 0p1n5t 0p1n5t 0p0n5t 0p1n5t -- TRDX4-20 0p0n3t 0p1n3t 0p1n3t 0p1n3t -- 0p1n3t 0p1n3t 0p1n3t 0p1n3t -- TRDX4-21 1p1n5t 0p0n5t 0p0n5t 0p0n5t -- 1p0n5t 0p0n5t 0p0n5t 1p0n5t -- TRDX4-22 0p0n3t 1p0n3t 0p0n3t 0p0n3t -- 0p1n3t 0p0n3t 0p0n3t 0p1n3t -- TRDX4-23 0p0n5t 1p0n5t 0p0n5t 1p0n5t -- 0p1n5t 0p0n5t 0p0n5t 0p1n5t -- TRDX4-25 1p0n5t 0p0n5t 0p0n5t 2p0n5t -- 0p1n5t 0p0n5t 0p0n5t 0p0n5t -- TRDX4-26 0p0n5t 0p0n5t 0p2n5t 0p1n5t -- 0p2n5t 0p1n5t 0p3n5t 0p0n5t -- TRDX4-27 0p0n7t 0p0n7t 0p1n7t 0p0n7t -- 0p1n7t 0p1n7t 0p0n7t 0p0n7t -- TRDX4-5T 0p0n3t 0p0n3t 0p1n3t 0p0n3t -- 0p1n3t 0p0n3t 0p1n3t 0p1n3t -- TRDX4-6T -- 0p1n5t 0p4n5t 0p0n5t -- 0p3n5t 0p1n5t 0p4n5t 0p2n5t -- TRDX4-7T 1p0n2t 0p0n2t 0p0n2t 0p0n2t -- 0p0n2t 0p1n2t 0p0n2t 0p0n2t --
TABLE-US-00005 TABLE 5 Summary of transgenic corn plants with altered phenotypes in AGH nitrogen-deficit screens Gene_ID AntA AntS Bmass ClrpC ClrpS Cnop PlntH WUE WtrAp WtrCt TRDX4-01 3p0n5t 0p1n5t 0p1n5t 0p0n5t -- 0p3n5t 0p2n5t 0p2n5t 1p2n5t -- TRDX4-02 0p1n5t 0p1n5t 5p0n5t 3p0n5t -- 4p0n5t 3p1n5t 5p0n5t 5p0n5t -- TRDX4-03 -- 0p0n5t 0p3n5t 0p0n5t -- 0p4n5t 0p2n5t 0p2n5t 0p3n5t -- TRDX4-04 0p0n5t 0p0n5t 0p0n5t 1p0n5t -- 0p0n5t 0p2n5t 0p0n5t 1p0n5t -- TRDX4-05 5p0n5t 0p1n5t 0p2n5t 0p0n5t -- 0p1n5t 0p0n5t 0p3n5t 0p4n5t -- TRDX4-07 -- 0p0n5t 1p0n5t -- 0p1n5t 0p0n5t 0p1n5t 1p0n5t 1p0n5t 0p0n5t TRDX4-09 0p0n5t 0p0n5t 0p0n5t 0p0n5t -- 0p0n5t 0p0n5t 0p0n5t 0p1n5t -- TRDX4-11 0p0n5t 0p0n5t 0p2n5t 0p1n5t -- 0p0n5t 0p0n5t 0p0n5t 0p3n5t -- TRDX4-12 0p4n5t 0p0n5t 1p0n5t 1p0n5t -- 2p0n5t 0p0n5t 1p0n5t 0p0n5t -- TRDX4-13 0p2n5t 0p0n5t 4p0n5t 0p1n5t -- 1p0n5t 3p0n5t 3p0n5t 3p0n5t -- TRDX4-14 3p0n5t 0p0n5t 0p2n5t 0p0n5t -- 0p1n5t 0p4n5t 0p1n5t 0p3n5t -- TRDX4-16 1p0n5t 0p0n5t 0p2n5t 0p0n5t -- 0p1n5t 1p2n5t 0p2n5t 0p1n5t -- TRDX4-17 0p2n5t 1p0n5t 0p0n5t 1p1n5t -- 2p0n5t 0p1n5t 0p0n5t 0p4n5t -- TRDX4-18 0p3n5t 0p1n5t 2p0n5t 0p0n5t -- 1p0n5t 0p0n5t 3p0n5t 0p1n5t -- TRDX4-19 0p1n3t 0p0n3t 1p0n3t 0p0n3t -- 2p0n3t 1p0n3t 0p0n3t 2p0n3t -- TRDX4-19x 0p0n5t 0p2n5t 0p0n5t 0p0n5t -- 0p1n5t 0p0n5t 1p0n5t 0p1n5t -- TRDX4-1T 1p0n5t 0p1n5t 1p1n5t 0p0n5t -- 0p1n5t 0p2n5t 1p1n5t 1p0n5t -- TRDX4-20 0p2n3t 0p2n3t 2p0n3t 0p0n3t -- 1p0n3t 1p0n3t 2p0n3t 2p0n3t -- TRDX4-21 0p2n5t 0p0n5t 1p1n5t 2p0n5t -- 1p1n5t 0p1n5t 2p0n5t 0p3n5t -- TRDX4-22 0p2n3t 0p0n3t 2p0n3t 0p0n3t -- 1p0n3t 2p0n3t 2p0n3t 3p0n3t -- TRDX4-23 0p0n5t 0p0n5t 1p0n5t 0p0n5t -- 1p0n5t 1p0n5t 1p0n5t 1p0n5t -- TRDX4-25 0p0n5t 0p0n5t 0p0n5t 1p0n5t -- 0p1n5t 0p2n5t 1p0n5t 0p2n5t -- TRDX4-26 0p0n5t 0p2n5t 0p0n5t 4p0n5t -- 0p1n5t 0p5n5t 0p0n5t 0p5n5t -- TRDX4-27 1p1n7t 0p1n7t 0p0n7t 1p0n7t -- 0p1n7t 0p1n7t 0p0n7t 1p1n7t -- TRDX4-5T 0p0n3t 0p0n3t 0p1n3t 0p0n3t -- 0p2n3t 0p1n3t 0p2n3t 0p3n3t -- TRDX4-6T -- 0p2n5t 5p0n5t 5p0n5t -- 3p0n5t 4p0n5t 5p0n5t 5p0n5t -- TRDX4-7T 0p1n3t 0p1n3t 3p0n3t 0p0n3t -- 3p0n3t 0p0n3t 3p0n3t 2p0n3t --
TABLE-US-00006 TABLE 6 Summary of transgenic corn plants with altered phenotypes in AGH water-deficit screens Gene_ID AntA AntS Bmass ClrpC ClrpS Cnop PlntH WUE WtrAp WtrCt TRDX4-01 0p0n5t 1p1n5t 1p0n5t 1p0n5t -- 1p1n5t 1p1n5t 0p0n5t 1p0n5t -- TRDX4-02 0p0n5t 3p0n5t 2p0n5t 0p0n5t -- 0p0n5t 1p1n5t 1p1n5t 0p3n5t -- TRDX4-03 -- 0p1n5t 0p0n5t 1p0n5t -- 1p0n5t 0p0n5t 0p1n5t 1p0n5t -- TRDX4-04 0p2n5t 0p0n5t 1p0n5t 0p0n5t -- 1p0n5t 0p2n5t 1p0n5t 0p0n5t -- TRDX4-05 1p0n5t 0p0n5t 0p1n5t 0p0n5t -- 0p1n5t 1p0n5t 0p1n5t 1p2n5t -- TRDX4-07 -- 4p0n5t 0p1n5t -- 3p1n5t 0p2n5t 0p2n5t 0p1n5t 0p4n5t 4p1n5t TRDX4-09 1p0n5t 0p0n5t 0p0n5t 0p1n5t -- 0p1n5t 0p1n5t 0p0n5t 1p4n5t -- TRDX4-11 2p0n5t 0p0n5t 0p0n5t 2p0n5t -- 0p0n5t 0p0n5t 0p0n5t 1p2n5t -- TRDX4-12 1p1n5t 0p1n5t 1p1n5t 0p0n5t -- 2p1n5t 2p0n5t 1p1n5t 2p0n5t -- TRDX4-13 4p0n5t 0p1n5t 0p5n5t 0p0n5t -- 0p5n5t 0p5n5t 0p3n5t 0p5n5t -- TRDX4-14 0p0n5t 0p0n5t 0p1n5t 0p1n5t -- 0p1n5t 1p1n5t 0p0n5t 0p2n5t -- TRDX4-16 0p0n5t 0p0n5t 0p0n5t 0p0n5t -- 0p0n5t 1p0n5t 0p0n5t 0p0n5t -- TRDX4-17 0p1n5t 0p0n5t 4p0n5t 2p0n5t -- 2p0n5t 4p0n5t 1p0n5t 4p0n5t -- TRDX4-18 0p2n5t 0p0n5t 3p0n5t 1p0n5t -- 1p0n5t 1p2n5t 2p1n5t 4p0n5t -- TRDX4-19 0p1n4t 1p0n4t 0p2n4t 0p0n4t -- 1p1n4t 0p2n4t 0p1n4t 0p1n4t -- TRDX4-19x 0p1n5t 0p0n5t 0p0n5t 0p0n5t -- 0p0n5t 0p0n5t 0p0n5t 1p2n5t -- TRDX4-1T 0p1n2t 0p0n2t 1p0n2t 0p0n2t -- 0p0n2t 0p0n2t 1p0n2t 0p0n2t -- TRDX4-20 0p1n3t 0p0n3t 0p0n3t 0p0n3t -- 0p0n3t 0p1n3t 0p0n3t 0p0n3t -- TRDX4-21 0p1n5t 0p0n5t 3p1n5t 2p0n5t -- 3p0n5t 2p0n5t 1p0n5t 4p1n5t -- TRDX4-22 0p0n1t 0p0n1t 1p0n1t 0p1n1t -- 0p0n1t 0p0n1t 0p0n1t 1p0n1t -- TRDX4-23 0p3n5t 0p1n5t 4p0n5t 1p0n5t -- 4p0n5t 0p0n5t 1p0n5t 4p0n5t -- TRDX4-25 1p0n5t 1p0n5t 1p0n5t 1p0n5t -- 1p0n5t 0p0n5t 0p0n5t 0p1n5t -- TRDX4-26 1p0n5t 3p0n5t 5p0n5t 0p0n5t -- 5p0n5t 2p0n5t 3p0n5t 1p0n5t -- TRDX4-27 4p0n7t 0p0n7t 0p2n7t 1p1n7t -- 1p0n7t 0p2n7t 0p1n7t 0p1n7t -- TRDX4-5T 2p0n3t 0p0n3t 0p3n3t 0p0n3t -- 0p3n3t 0p3n3t 0p3n3t 0p3n3t -- TRDX4-6T -- 0p1n5t 0p0n5t 0p0n5t -- 0p0n5t 1p0n5t 0p0n5t 0p0n5t -- TRDX4-7T 0p0n3t 0p0n3t 0p0n3t 0p1n3t -- 3p0n3t 0p0n3t 1p0n3t 0p0n3t --
Example 4. Phenotypic Evaluation of Transgenic Plants for Increased Nitrogen Use Efficiency, Increased Water Use Efficiency and Increased Yield
[0178] Corn field trials were conducted to identify genes that can improve nitrogen use efficiency (NUE) under nitrogen limiting conditions leading to increased yield performance as compared to non transgenic controls. For the Nitrogen field trial results shown in Tables 7 and 9, each field was planted under nitrogen limiting condition (60 lbs/acre) and corn ear weight or yield was compared to non transgenic control plants.
[0179] Corn field trials were conducted to identify genes that can improve water use efficiency (WUE) under water limiting conditions leading to increased yield performance as compared to non transgenic controls. The water use efficiency trials for results shown in Tables 7 and 9 were conducted under managed water limiting conditions, and the corn ear weight or yield was compared to non transgenic control plants.
[0180] Corn and soybean field trials were conducted to identify genes that can improve broad-acre yield (BAY) under standard agronomic practice. The broad-acre yield trials for results shown in Tables 7 and 9 were conducted under standard agronomic practice, and the corn or soybean yield was compared to non transgenic control plants.
[0181] Table 7 provides a list of genes for producing transgenic plants with increased nitrogen use efficiency (NUE), increased water use efficiency (WUE), and increased broad-acre yield (BAY) as compared to a control plant. Polynucleotide sequences in constructs with at least one event showing significant yield or ear weight increase across multiple locations at p.ltoreq.0.2 are included. The genes were expressed with constitutive promoters unless noted otherwise under "Specific Expression Pattern". Promoter of specific expression pattern was chosen over constitutive promoter, based on the understanding of the gene function, or based on the observed lack of significant yield increase when the gene was expressed with constitutive promoter. The elements of Table 7 are described by reference to:
[0182] "Crop" which refers to the crop in trial, which is either corn or soybean;
[0183] "Condition" which refers to the type of field trial, which is BAY for broad acre yield trial under standard agronomic practice (SAP), WUE for water use efficiency trial, and NUE for nitrogen use efficiency trial;
[0184] "Specific Expression Pattern" which refers to the expected expression pattern or promoter type, instead of constitutive;
[0185] "Gene ID" which refers to the gene identifier as defined in Table 1;
[0186] "Yield results" which refers to the recombinant DNA in a construct with at least one event showing significant yield increase at p.ltoreq.0.2 across locations. The first number refers to the number of events with significant yield or ear weight increase, whereas the second number refers to the total number of events tested for each recombinant DNA in the construct.
TABLE-US-00007 TABLE 7 Recombinant DNA with protein-coding genes for increased nitrogen use efficiency, increased water use efficiency and increased yield Specific Expression Yield Crop Condition Pattern Gene ID Results corn BAY leaf preferred TRDX4-01 2/15 corn BAY seed preferred TRDX4-02 1/13 corn BAY TRDX4-03 2/8 corn BAY TRDX4-04 1/23 corn BAY TRDX4-05 1/23 corn BAY TRDX4-06 1/7 corn WUE TRDX4-07 2/4 corn BAY cold inducible TRDX4-08 4/26 corn BAY TRDX4-09 1/25 corn NUE TRDX4-10 1/12 corn BAY TRDX4-11 1/20 corn BAY TRDX4-12 8/20 corn BAY TRDX4-13 2/32 corn BAY TRDX4-14 2/20 soybean BAY TRDX4-15 4/13 corn BAY TRDX4-16 1/20 corn BAY leaf preferred TRDX4-17 2/13 corn BAY TRDX4-18 2/19 corn WUE leaf preferred TRDX4-19 1/5 corn BAY TRDX4-19 2/22 corn BAY leaf preferred TRDX4-20 1/20 corn BAY TRDX4-21 4/19 corn NUE TRDX4-22 2/10 corn BAY leaf preferred TRDX4-23 1/19 soybean BAY TRDX4-24 3/15 corn BAY TRDX4-25 2/18 corn BAY root preferred TRDX4-26 3/21 corn BAY TRDX4-27 3/24
[0187] Table 8 provides a list of polynucleotide sequences of promoters with specific expression patterns. To convey the specific expression patterns, choices of promoters are not limited to those listed in Table 8.
TABLE-US-00008 TABLE 8 Promoter sequences and expression patterns Nucleotde SEQ Promoter Expression ID NO. Pattern 97 Cold inducible 98 Seed preferred 99 Leaf preferred 100 Leaf preferred
[0188] Table 9 provides a list of suppression target genes and miRNA construct elements provided as recombinant DNA for production of transgenic corn or soybean plants with increased nitrogen use efficiency, increased water use efficiency and increased yield. The elements of Table 9 are described by reference to:
[0189] "Crop" which refers to the crop in trial, which is either corn or soy;
[0190] "Condition" which refers to the type of field trial, which is BAY for broad acre yield trial under standard agronomic practice, WUE for water use efficiency trial, and NUE for nitrogen use efficiency trial;
[0191] "Target Gene ID" which refers to the suppression target gene identifier as defined in Table 2;
[0192] "Engineered miRNA precursor SEQ ID NO." which identifies a nucleotide sequence of the miRNA construct;
[0193] "Yield results" which refers to the recombinant DNA in a construct with at least one event showing significant yield increase at p.ltoreq.0.2 across locations. The first number refers to the number of events with significant yield or ear weight increase, whereas the second number refers to the total number of events tested for each sequence in the construct.
TABLE-US-00009 TABLE 9 miRNA Recombinant DNA constructs suppressing targeted genes for increased nitrogen use efficiency, increased water use efficiency and increased yield Engineered Target miRNA precursor Yield Crop Condition Gene ID SEQ ID NO. Results corn BAY TRDX4-1T 67 1/13 soybean BAY TRDX4-3T 68 4/15 soybean BAY TRDX4-4T 69 3/14 corn BAY TRDX4-5T 70 2/20 corn NUE TRDX4-6T 71 1/6 corn WUE TRDX4-6T 71 1/6 corn BAY TRDX4-6T 71 2/20 corn WUE TRDX4-7T 72 1/5
Example 5. Homolog Identification
[0194] This example illustrates the identification of homologs of proteins encoded by the DNA identified in Table 1 which were used to provide transgenic seed and plants having enhanced agronomic traits. From the sequences of the homolog proteins, corresponding homologous DNA sequences can be identified for preparing additional transgenic seeds and plants with enhanced agronomic traits.
[0195] An "All Protein Database" was constructed of known protein sequences using a proprietary sequence database and the National Center for Biotechnology Information (NCBI) non-redundant amino acid database (nr.aa). For each organism from which a polynucleotide sequence provided herein was obtained, an "Organism Protein Database" was constructed of known protein sequences of the organism; it is a subset of the All Protein Database based on the NCBI taxonomy ID for the organism.
[0196] The All Protein Database was queried using amino acid sequences provided in Table 1 using NCBI "blastp" program with E-value cutoff of 1e-8. Up to 1000 top hits were kept, and separated by organism names. For each organism other than that of the query sequence, a list was kept for hits from the query organism itself with a more significant E-value than the best hit of the organism. The list contains likely duplicated genes of the polynucleotides provided herein, and is referred to as the Core List. Another list was kept for all the hits from each organism, sorted by E-value, and referred to as the Hit List.
[0197] The Organism Protein Database was queried using polypeptide sequences provided in Table 1 using NCBI "blastp" program with E-value cutoff of 1e-4. Up to 1000 top hits were kept. A BLAST searchable database was constructed based on these hits, and is referred to as "SubDB". SubDB is queried with each sequence in the Hit List using NCBI "blastp" program with E-value cutoff of 1e-8. The hit with the best E-value was compared with the Core List from the corresponding organism. The hit is deemed a likely ortholog if it belongs to the Core List, otherwise it is deemed not a likely ortholog and there is no further search of sequences in the Hit List for the same organism. Homologs with at least 95% identity over 95% of the length of the polypeptide sequences provided in Table 1 are reported below in Tables 10 and 11.
[0198] Table 10 provides a list of homolog genes, of which the elements are described by reference to:
[0199] "PEP SEQ ID NO." which identifies an amino acid sequence.
[0200] "Homolog ID" which refers to an alphanumeric identifier, the numeric part of which is the NCBT Genbank GI number.
[0201] "Gene Name and Description" which is a common name and functional description of the gene.
TABLE-US-00010 TABLE 10 Homolog genes information PEP SEQ ID NO. Homolog ID Gene Name and Description 79 gi_735918 gi | 735918 | emb | CAA84367.1 | asparaginase [Arabidopsis thaliana] 80 gi_110742427 gi | 110742427 | dbj | BAE99132.1 | cyclic nucleotide-gated cation channel [Arabidopsis thaliana] 81 gi_215261267 gi | 215261267 | pdb | 3EBL | A Chain A, Crystal Structure Of Rice Gid1 Complexed With Ga4 [Oryza sativa Japonica group] 82 gi_193211383 gi | 193211383 | ref | NP_001105952.1 | plastid phosphate/phosphoenolpyruvate translocator1 [Zea mays] 83 gi_3164136 gi | 3164136 | dbj | BAA28535.1 | cytochrorne P450 monooxygenase [Arabidopsis thaliana] 84 gi_28867617 gi | 28867617 | ref | NP_790236.1 | phosphoglycerate kinase [Pseudomonas syringae pv. tomato str. DC3000] 85 gi_71734219 gi | 71734219 | ref | YP_276916.1 | phosphoglycerate kinase [Pseudomonas syringae pv. phaseolicola 1448A] 86 gi_66048014 gi | 66048014 | ref | YP_237855.1 | phosphoglycerate kinase [Pseudomonas syringae pv. syringae 8728a] 87 gi_242053823 gi | 242053823 | ref | XP_002456057.1 | hypothetical protein SORBIDRAFT_03g029630 [Sorghum bicolor] 88 gi_21593232 gi | 21593232 | gb | AAM65181.1 | unknown [Arabidopsis thaliana] 89 gi_226510490 gi | 22651.0490 | ref | NP_001148910.1 | LOC100282530 [Zea mays] gi | 195623174 | gb | ACG33417.1 | pre-mRNA-splicing factor 1SY1 [Zea mays] 90 gi_242065688 gi | 242065688 | ref | XP_002454133.1 | hypothetical protein SORBIDRAFT_04g025200 [Sorghum bicolor] 91 gi_21593552 gi | 21593552 | gb | AAM65519.1 | unknown [Arabidopsis thaliana] 92 gi_21593833 gi | 21593833 | gb | AAM65800.1 | glutaredoxin-like protein [Arabidopsis thaliana] 93 gi_2265066.54 gi | 226506654 | ref | NP_001146301.1 | DNA mismatch repair protein MSH2 [Zea mays] 94 gi_242050756 gi | 242050756 | ref | XP_002463122.1 | hypothetical protein SORBIDRAFT_02g038230 [Sorghum bicolor] 95 gi_255639875 gi | 255639875 | gb | ACU20230.1 | unknown [Glycine max] 96 gi_195623972 gi | 195623972 | gb | ACG33816.1 | triose phosphate/phosphate translocator, non-green plastid, chloroplast precursor [Zea mays]
[0202] Table 11 describes the correspondence between the protein-coding genes in Table 1, suppression target genes in Table 2, and their homologs, and the level of protein sequence alignment between the gene and its homolog. Note that homologs can be from Table 1, 2 or 10.
TABLE-US-00011 TABLE 11 Correspondence of Genes and Homologs Percent Percent Gene Homolog Percent Gene ID Homolog ID Coverage Coverage Identity TRDX4-02 gi_735918 100 100 99 TRDX4-03 gi_110742427 100 100 99 TRDX4-05 gi_215261267 100 97 100 TRDX4-06 TRDX4-1T 100 100 99 TRDX4-06 gi_193211383 100 100 99 TRDX4-08 gi_3164136 100 100 99 TRDX4-09 gi_28867617 99 100 100 TRDX4-09 gi_71734219 99 100 96 TRDX4-09 gi_66048014 99 100 96 TRDX4-10 gi_242053823 100 100 95 TRDX4-11 gi_21593232 100 100 99 TRDX4-12 gi_226510490 100 100 99 TRDX4-12 gi_242065688 100 100 98 TRDX4-14 gi_21593552 100 100 99 TRDX4-18 gi_21593833 99 100 100 TRDX4-22 gi_226506654 100 100 99 TRDX4-22 gi_242050756 100 100 95 TRDX4-25 gi_255639875 100 100 99 TRDX4-1T TRDX4-06 100 100 99 TRDX4-1T gi_195623972 100 100 99 TRDX4-1T gi_193211383 100 100 99
Example 6. Use of Suppression Methods to Suppress Expression of Target Genes
[0203] This example illustrates monocot and dicot plant transformation with recombinant DNA constructs that are useful for stable integration into plant chromosomes in the nuclei of plant cells to provide transgenic plants having enhanced traits by suppression of the expression of target genes.
[0204] Various recombinant DNA constructs for use in suppressing the expression of a target gene in transgenic plants are constructed based on the nucleotide sequence of the gene encoding the protein that has an amino acid sequence selected from the group consisting of SEQ ID NOs: 61-66, where the DNA constructs are designed to express (a) a miRNA that targets the gene for suppression, (b) an RNA that is a messenger RNA for a target protein and has a synthetic miRNA recognition site that results in down modulation of the target protein, (c) an RNA that forms a dsRNA and that is processed into siRNAs that effect down regulation of the target protein. (d) a ssRNA that forms a transacting siRNA which results in the production of siRNAs that effect down regulation of the target protein.
[0205] Each of the various types of recombinant DNA constructs is used in transformation of a corn cell using the vector and method of Examples 1 and 2 to produce multiple events of transgenic corn cell. Such events are regenerated into transgenic corn plants and are screened to confirm the presence of the recombinant DNA and its expression of RNA for suppression of the target protein. The population of transgenic plants from multiple transgenic events are also screened to identify the transgenic plants that exhibit altered phenotype or enhanced trait.
Sequence CWU
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1
1041165DNAArabidopsis thaliana 1atggtgaaca acgttgtctc tatcgaaaag
atgaaagcac tctggcactc cgaggttcat 60gatgaacaaa aatgggcggt gaacatgaaa
cttctgcgag cacttggtat gtttgcagga 120ggagtcgtcc tcatgcgtag ctatggggat
ctcatgggag tttga 1652948DNAArabidopsis thaliana
2atggtggggt gggcgattgc gctacacggc ggtgccggag acattccgat cgatctcccc
60gacgagcgac gtatccctcg tgagagcgcc ctccgtcact gcctcgatct tggcatctcc
120gccctcaaat ccggcaagcc tcccttggac gtcgccgaac ttgtcgttcg tgaacttgag
180aaccacccgg acttcaatgc gggtaaagga tctgtcttaa ctgcacaagg cactgttgaa
240atggaagctt ccattatgga cggtaaaacc aaaagatgtg gagctgtctc cggcttgacc
300actgttgtta atcccatttc tttagctcgc ctcgtcatgg agaaaactcc tcatatatat
360cttgcattcg atgctgctga agcttttgca agagcacatg gtgttgagac ggtagattct
420agccatttca taactcctga aaacattgca aggctaaagc aggccaaaga attcaatcga
480gtccagttgg attacacagt ccctagtccg aaagtaccgg acaattgcgg tgacagccaa
540ataggaacgg tcggatgtgt agctgtggac agtgctggaa atctagcttc ggctacatca
600acgggcggtt atgtcaacaa aatggttggc agaattgggg atacgccagt cattggcgca
660ggaacttacg ctaaccacct ttgtgccatc tcagccacag gtaaaggaga ggatatcatc
720cgtggaaccg tggctagaga cgtggctgca ctcatggaat ataaaggctt gtctttgact
780gaggcagcgg cttatgttgt tgaccaatct gttcccagag gaagctgtgg actcgttgct
840gtctctgcca atggtgaagt cacaatgccg tttaacacta ccggaatgtt cagggcttgt
900gctagcgaag atggttactc tgagatcgca atctggccaa acaattga
94832181DNAArabidopsis thaliana 3atgccctctc accccaactt catcttcagg
tggattggac tgttttccga taagttccgt 60cgacaaacga ctgggatcga tgaaaacagt
aacctccaaa tcaacggtgg agattcgagc 120agcagcggca gcgatgagac gccggtgcta
agctccgtcg agtgttacgc ttgcacacaa 180gtaggcgtcc cagctttcca ttcaactagc
tgcgatcaag ctcacgcgcc ggagtggcgt 240gcctccgccg gctcttctct agttccgatc
caggaaggat ctgtccctaa cccagcccga 300accagattcc gacgtctcaa aggtccgttt
ggtgaagttc tcgatcctag gagcaagcgc 360gtgcagagat ggaaccgcgc gttgctttta
gctcgtggga tggctttagc ggtggatccg 420ctcttcttct acgcgctttc catcggccga
actaccggac cggcgtgtct ttacatggat 480ggtgcgttcg ccgcggtggt cacggtgctc
cgcacgtgtc tcgatgctgt tcatctttgg 540cacgtgtggc ttcaattcag actggcctac
gtctcgagag agtcgcttgt cgttggttgt 600gggaagctcg tttgggatcc acgcgccatc
gcgtctcact acgcacgctc tctcactggc 660ttctggtttg atgttatcgt catcctccct
gtccctcagg cagtgttttg gttagttgtg 720ccgaaactga taagagaaga gaaggttaag
ctgataatga cgattctgct gctaatattc 780ttgttccagt tcctccccaa gatttatcac
tgcatctgtt tgatgagaag gatgcagaag 840gtcactggtt acatttttgg aactatttgg
tggggttttg ctcttaatct catcgcatat 900ttcatcgctt ctcatgttgc tgggggatgt
tggtatgttc tcgcaataca gcgtgttgct 960tcttgcataa gacaacaatg tatgagaacc
gggaactgca atctgagtct ggcttgcaaa 1020gaagaggtct gttaccaatt tgtgtcaccg
acaagcacag ttggatatcc atgcttatct 1080ggaaacctta ccagtgtggt caataagcct
atgtgcttag actctaacgg accattccga 1140tatggtatct accgttgggc acttccagtc
atctccagca actctcttgc ggttaagatc 1200ctttacccca tcttctgggg cctaatgact
ctcagcacat ttgcgaatga tcttgagccc 1260acaagcaact ggctcgaggt tattttcagt
atagttatgg ttctaagtgg cttgttactt 1320ttcacgctgt tgataggaaa cattcaggtg
tttttgcatg cggtaatggc gaaaaaaagg 1380aaaatgcaga tacggtgtag ggatatggaa
tggtggatga aacgtaggca gttaccttcc 1440cggttaagac agagggttag gcgatttgag
cggcagagat ggaatgcctt gggtggtgaa 1500gacgagctag aacttataca tgatttgcct
ccgggtcttc gaagagatat caaacgatat 1560ctttgctttg atctcattaa caaggtgcca
ttgttcaggg gcatggacga cttgatcctc 1620gacaacattt gcgatcgggc taagcctcga
gtcttctcta aagacgaaaa gatcatccgt 1680gaaggagatc ctgtacagag aatgatattc
atcatgcgtg gacgagtcaa acgtatacag 1740agcctaagca aaggcgtcct agccactagt
acactagaac caggcggtta cttgggcgac 1800gagctactct catggtgcct acgtcgcccg
tttctggacc gtcttccccc ttcctcagca 1860acatttgtct gcctagaaaa catcgaggca
ttctccctcg gatccgaaga tcttaggtac 1920attaccgatc atttccgtta taaattcgcg
aacgagcggc ttaagcggac cgcaagatac 1980tattcctcaa actggaggac gtgggcagcg
gtaaatattc agatggcgtg gcgccggcgt 2040aggaaaagaa cccgtggtga aaacatcggc
ggttcgatga gtcctgtgtc ggagaatagc 2100attgaaggta acagtgaacg ccggttactt
cagtatgcag ctatgttcat gtccattcga 2160ccgcatgatc atctcgaata a
218141404DNAArabidopsis thaliana
4atgattcttg atttgggttt tccttgtttt gttcctcctc gaaccagctc tcgtgaggac
60aacaaagctt ggcttctggc tgaaacagag ccgaagctta ttgactcaga acaacattcg
120ttgcagtctt cgtttaggtt tagtctttgc tcacagttgg agctggagaa gattaaaaag
180gagaaacctt cgttgtctta tcggaatttt ccagtgtctg aaggatcaga gacggttctg
240ctagtgaatc tggagaatga gacaggagaa ttgacaggtg agatgaattg gtcgagaggc
300ctttcactgg agaagagtat ttctccggtg gccgattctt tgatccgatt cagttaccgc
360gaactcctca ctgccacgcg caatttctca aaacggaggg ttttgggaag aggagcttgt
420agctatgttt ttaagggaag aatcgggatt tggcgtaaag ccgtggccat caaaagactt
480gataaaaaag ataaagaatc tccaaagtcg ttttgcagag agttgatgat tgcaagctct
540cttaatagcc ccaacgttgt gcctctgcta ggtttctgta tcgatcccga tcaagggctt
600ttcttggtgt acaagtatgt gtctggtggc agcctcgaac gctttttaca tgataagaag
660aaaaagaaga gtaggaagac ccccttgaat ctgccttggt ctacaaggta caaggttgcc
720ttaggtattg cagatgccat agcctattta cataatggca ctgagcaatg cgttgtgcat
780agagacatta aaccctcaaa tattcttctt tcctcaaaca aaattccaaa gttgtgtgat
840tttgggttgg ctacttggac cgctgcgcct tcggttcctt tcctctgtaa aaccgtgaaa
900ggaacttttg gttatctggc tcctgagtat ttccaacacg gcaagatatc tgacaagacc
960gatgtttacg catttggggt cgtgttgctt gagctaataa ctggtcggaa gccaattgaa
1020gcaagaagac catctggtga agaaaatttg gtagtttggg caaaaccgtt gttgcataga
1080gggatagaag ctacagagga gttgctagat ccaaggctga aatgtactag aaaaaactcg
1140gcttcgatgg agcgtatgat ccgagctgcg gcagcgtgtg tgatcaatga ggaatcacga
1200agaccgggga tgaaggagat actttcaatc ctaaaaggcg gtgaagggat agaactaagg
1260acgttatcaa gccggaagaa atcaaatctt ccgggtataa tggactgtta tccgcagttg
1320caacggacaa aatctgagat gaagagtcat cttacgcttg cgatgctcgg agtaacggaa
1380tttgaagctg atgatctttt gtag
140451065DNAOryza sativa 5atggccggca gcgacgaggt caaccgcaac gagtgcaaga
cggtggtgcc gctccacaca 60tgggtgctca tctccaactt caagctgtcg tacaacattc
tgcggcgggc ggacgggacg 120ttcgagcggg acctcgggga gtacctggac aggagggtgc
cggcgaacgc gcggccgctg 180gagggggtgt cgtcgttcga ccacatcatc gaccagtcgg
tggggctgga ggtgcgcatc 240taccgggcgg cggcggaggg tgacgcggag gagggggcgg
cggcggtgac gcggcccatc 300cttgagttcc tgacggacgc gccagcggcg gagccgttcc
cggtgatcat attcttccac 360ggcggcagct tcgtgcactc gtcggccagc tcgaccatct
acgacagtct gtgccgccgg 420ttcgtgaagc tgagcaaggg cgtcgtggtg tccgtcaact
accggcgcgc gccggagcac 480cgctacccgt gcgcgtacga cgacgggtgg accgcgctca
agtgggtcat gtcgcagccg 540ttcatgcgca gcggcggcga cgcgcaggcc cgcgtgttcc
tctccggcga cagctccggc 600ggcaacatcg cccaccacgt cgccgtccgc gccgccgacg
agggcgtcaa ggtctgcggc 660aacatcctgc tcaacgccat gttcggcggc accgagcgca
cggagtcgga gcggcggctc 720gacggcaagt acttcgtgac gctccaggac agggactggt
actggaaggc gtacctgccg 780gaggacgccg accgggacca tccggcgtgc aacccgttcg
gcccgaacgg ccggcggctc 840gggggcctcc ccttcgccaa gagcctcatc atcgtgtcgg
gcctggacct cacctgcgac 900cggcagctcg cctacgccga cgccctccgg gaggacggcc
accacgtcaa ggttgtccaa 960tgcgagaacg ccacggtggg gttctacctg ttgcccaaca
ccgtccacta ccacgaggtc 1020atggaggaga tctccgactt cctcaacgct aacctctact
actag 106561173DNAZea mays 6atgcagagcg cggctgccat
cgggctccta cggccatgtg ccgcgcggcc gctcgccgcc 60tacactagcc cacgccgcgg
cgccggcgcg tgcagcggcg gcacccagcc gatcatcacg 120ccccgcggca tccgcctctc
cgcccgcccc ggtctcgtgc cggcctcgcc gctggaggag 180aaggagaacc ggagatgcag
ggccagtatg cacgcggcgg cgtcggccgg agaggaagct 240gggggagggc tcgccaagac
gctgcagctg ggggcgcttt tcgggctctg gtacctcttc 300aacatctact tcaacatcta
caacaagcag gttctgaagg ttttgccata ccctataaac 360atcacaacgg tgcagtttgc
tgttggaagt gccattgctt tgttcatgtg gatcactggt 420atccataaaa ggccaaagat
ttcgggtgcc cagcttttcg ctatccttcc tctagctatt 480gtccatacca tgggcaatct
tttcacaaac atgagccttg gaaaggtggc agtgtcattt 540acacatacta taaaggccat
ggaacctttc ttctcagttc tcctttcagc aattttcctt 600ggggagttgc ctacgccatg
ggttgtgttg tctcttcttc cgattgttgg tggtgtagct 660ttggcatccc ttactgaggc
ctcctttaac tgggctggat tttggagtgc aatggcttca 720aatgtaacct tccagtcaag
gaatgtgcta agcaagaaac ttatggtgaa gaaagaggaa 780tctctcgaca acattaacct
attctcgatc attacagtca tgtcattctt cctgttggcc 840ccagtaacct tacttacaga
aggtgttaaa gttagtccag cagtgttgca gtctgctggt 900ttgaacttga aacaggtata
cacaaggtca ttgattgctg cattctgctt ccatgcatac 960caacaggtgt catacatgat
cctcgccagg gtatccccag tcacacattc agtgggcaat 1020tgcgtcaagc gtgtggtggt
cattgtgacc tctgttctgt tcttcaggac ccctgtttct 1080cccatcaact ctcttggtac
cgggatcgct cttgctggag ttttcctata ctcgcaattg 1140aagagactta agcccaagcc
caagactgct tag 117371434DNAArabidopsis
thaliana 7atggcgcatc tactttcagc ttcatgccct tcagttatct cacttagcag
cagcagcagc 60aagaattcag ttaagccgtt tgtttcaggg cagaccttct tcaatgctca
gcttctttca 120agatcttctc tcaaaggact tctcttccaa gagaagaaac cgagaaaaag
ctgcgttttc 180agagcaactg ctgtacctat aacccaacaa gcaccacccg aaacatctac
caataactca 240tcctctaaac caaagcgtgt tatggtcatt ggtggagatg gttattgcgg
ttgggctact 300gctctccact tgtccaagaa gaattacgaa gtttgcattg ttgacaacct
tgtaagacgt 360cttttcgacc accagcttgg acttgagtca ttgactccta ttgcctccat
tcatgaccga 420atcagccgat ggaaggcttt gacagggaaa tcaattgagt tgtacgttgg
tgatatctgt 480gatttcgaat tcttagctga gtctttcaag tcttttgagc cggattcagt
tgtccacttt 540ggggaacaga gatccgctcc ttactcgatg attgaccggt ccagagcagt
ttatacacag 600cacaacaatg tgattgggac tctcaacgtt ctctttgcta taaaagagtt
tggagaggag 660tgtcatcttg taaaacttgg gacgatgggt gagtatggaa ctccaaatat
tgacatcgag 720gaaggttata taaccataac ccacaacggt agaactgaca ctttgccata
ccccaagcaa 780gctagctcct tttatcatct tagcaaagtt catgattcgc acaacattgc
ttttacttgc 840aaggcttggg gtattagagc cactgatctc aaccaaggag ttgtttatgg
agtgaagact 900gatgagacag agatgcatga ggaactccgt aaccgactgg attacgatgc
tgtgtttggt 960acagcactta accggttctg tgtgcaagct gctgttggtc acccacttac
agtttatggt 1020aaaggtggtc agacgagagg ctacctcgat ataagagaca cggttcaatg
tgttgagatc 1080gctatagcaa acccggcaaa agctggtgag ttccgggtct tcaaccaatt
tacagaacag 1140ttttcagtca atgaactggc ttcactcgtc actaaagcgg gttcaaagct
tgggctagac 1200gtgaaaaaga tgacggtgcc taacccgaga gtggaggcag aagaacatta
ctacaacgca 1260aagcacacta agctgatgga acttggactt gagcctcact atctatctga
ctcacttctt 1320gattcgttgc tcaactttgc tgttcagttt aaagatcgtg tggacacgaa
acaaatcatg 1380cctagtgttt cctggaagaa gattggcgtc aagactaagt ccatgaccac
atag 143481515DNAArabidopsis thaliana 8atggtgagtc ttctatcttt
tttcttgctt ctactcgtcc ccattttctt cttgttaatc 60ttcaccaaga agatcaagga
gtcaaaacaa aatcttcctc ctggcccagc aaagcttccg 120atcatcggaa acctacacca
gctccaaggg ttgcttcata aatgtcttca cgatctctcc 180aagaaacacg gacctgtgat
gcatctccgt ctagggtttg ccccaatggt cgtaatttca 240tcaagtgaag cagctgaaga
agctcttaaa acacatgacc ttgagtgttg ttcaagacct 300atcactatgg cctcaagggt
tttttcgcgt aacggtaaag acatcggatt tggggtttac 360ggtgatgaat ggagagagct
gcgtaagctt tcggttcgcg aattctttag cgtgaaaaaa 420gttcaatcct tcaagtatat
tagagaggaa gagaatgact tgatgatcaa gaaactgaaa 480gaattggctt cgaagcaatc
tccggtggat ttgagcaaaa tcctctttgg tctcactgcg 540agtatcatat tcagaaccgc
ctttggacaa agtttctttg ataacaagca tgtcgatcag 600gaaagcatca aagaactgat
gtttgaatct ctgagcaata tgacttttag attctctgat 660tttttcccta ctgctggtct
taaatggttt ataggctttg tgtcaggcca acataagagg 720ctttacaacg tcttcaacag
ggttgatact ttttttaatc atatagttga tgatcatcac 780tcgaagaaag caactcaaga
tcgtcctgat atggtcgacg ctatcttaga tatgatagat 840aatgaacaac aatatgcatc
tttcaagctc accgttgatc atctcaaagg agtcctctca 900aatatatatc acgctggaat
tgacacaagc gccatcacct tgatttgggc gatggcagag 960ctcgttagaa acccgcgggt
aatgaagaaa gctcaagacg agatccgaac ttgcattgga 1020atcaaacagg aaggaagaat
catggaagaa gatcttgata agcttcaata cttgaagctt 1080gtggtgaaag aaaccttaag
actacaccca gcagctcctc ttttacttcc tcgagaaaca 1140atggctgata tcaagattca
aggctacgac attcctcaga aaagagctct tcttgttaat 1200gcatggtcta taggacgaga
tccggaatcc tggaaaaatc ctgaagagtt taacccggag 1260aggtttattg attgtcctgt
ggattacaag ggacatagct ttgagttgtt accatttggt 1320tctggtcgga gaatttgtcc
aggaatagct atggcgatcg caaccattga attggggctc 1380ttgaatttgc tctacttctt
tgattggaat atgcctgaga agaagaaaga tatggacatg 1440gaagaagctg gtgatctcac
tgttgataag aaagttcctc ttgagcttct gccagttatt 1500cgcatcagtt tgtag
151591170DNAPseudomonas
syringae pv. tomato str. DC3000 9atggtcatga ccgtcttgaa gatgaccgac
ctcgatctgc aaggtaaacg tgtactgatc 60cgcgaagacc tcaacgtccc gataaaggac
ggcgttgtca gcagcgatgc acgtattctt 120gcttcgctgc cgaccatcag gctggcgctg
gaaaaaggcg cggctgtcat ggtctgctcg 180caccttggcc gtccgaccga gggcgagttt
tctgctgaaa acagcctcaa gccggttgct 240gaatacctga gcaaggcatt gggtcgtgac
gttccgctgg tcgccgatta cctggacggc 300gttgacgtca aggcgggcga tatcgtgctg
ttcgagaacg ttcgcttcaa caagggcgag 360aaaaagaacg ccgacgagct ggcgcagaag
tacgcggccc tgtgcgacgt gttcgtgatg 420gacgcttttg gcaccgctca ccgcgctgaa
ggctcgaccc acggcgtggc caaatacgcc 480aaggttgccg ctgctggccc gttgctggct
gccgaactgg aagcgctggg caaggcgctg 540ggcgctccgg ctcagccaat ggctgctatt
gttgccggct ccaaagtgtc caccaagctg 600gacgtgctca acagcctgag cgcgatctgc
gatcagttga ttgttggcgg cgggattgcc 660aacacctttc tggctgcagc cggtcacaag
gtcggtaaat cgctttacga gccagacctg 720ctcgacaccg cgcgcgccat tgccgccaag
gtcagcgtgc cgttgccgac tgacgtggtg 780gttgccaagg aattcgccga gagtgccact
gcaaccgtca agctgatcgc cgatgtggcc 840gacgacgaca tgattctgga tattggtcca
cagactgctg cgcacttcgc cgaactgttg 900aaatcttccg ggactatcct gtggaacggt
ccggttggcg tgttcgaatt cgaccagttc 960ggtgaaggca ccaaaacgct ggccaaggcc
attgctgaaa gcaaagcgtt ctccatcgcg 1020ggtggtggcg acaccctggc cgcgatcgac
aagtacggtg tggcagatca gatttcctat 1080atttcgaccg gtggcggtgc gttcctcgaa
ttcgtggaag gcaaggttct gccagcggtt 1140gaaatgctcg aacaacgtgc cagggcctag
1170101188DNAZea mays 10atgagtctca
taagagggat gggcaacgtt gccaagagat ggaaagaact caatggcttg 60aattactgga
agggcctagt tgatccgctc gacctcgacc tccgtaggaa catcatcaac 120tacggtgagc
tctcccaggc aacctacacc gggctgaaca gggagagaag atcaaggtac 180gctgggtctt
gcctcttcaa ccgcagagac ttcctcagca gggtggatgt atcaaacccg 240aacctgtatg
agatcacgaa gttcatatac gcgatgtgca ctgtcagctt acctgacggg 300ttcatggtca
agtctctctc aaaggctgca tggagcaggc agtcgaattg gatggggttt 360gttgcagtag
ctacggacga gggcaaggaa ctgcttggga ggcgggacgt ggtggtggcg 420tggcgtggca
ccataaggat ggtagagtgg gtcgatgatc ttgatatttc cttggtgcct 480gcttcggaaa
tagttcttcc aggcagcgca gccaacccct gtgtgcatgg agggtggctt 540tcagtctaca
cgagtgctga tccagggtca cagtacaaca aagagagcgc aagacatcag 600gtgttaaacg
aggtgaaaag gatacaggat ctgtacaagc cagaggagac gagcatcacc 660ataacaggcc
acagcctagg agcagcactt gccaccatca acgcaaccga catcgtctcc 720aatggctaca
acaggagctg ctgccctgtg tccgcgttcg tattcgggag ccccagagtc 780ggaaaccctg
atttccagaa ggcgttcgac agcgcggcgg acctgaggct gctccgcgtc 840cggaactctc
ccgacgtggt ccccaaatgg ccaaagctag ggtacagcga tgtcggcaca 900gagctgatga
tcgacacagg agaatcgccg tacctgaagg cccctggaaa ccccctgaca 960tggcatgaca
tggagtgcta catgcacggg gtcgctgggg ctcaggggag cagcggaggg 1020ttcgagctgt
tggtcgatcg ggacgttgct ttggtgaaca agcatgaaga tgcgctgaga 1080aatgagttcg
ctgtcccacc gtcgtggtgg gtggtgcaga acaaaggtat ggtgaaaggc 1140aaggatggcc
ggtggcatct ggccgaccat gaggaggatg atgactag
1188111539DNAArabidopsis thaliana 11atggctactc ttttagccac tcctatcttc
tctcctttag cttcttctcc agcaaggaac 60cgtctttctt gctctaagat ccgtttcggt
tccaaaaatg ggaaaattct caattctgat 120ggtgcccaga agttgaatct ctcaaaattc
cgtaaacccg atggccaaag atttctacaa 180atgggttctt ctaaagagat gaactttgag
agaaaactct cagtccaagc tatggatggt 240gcaggaacag gaaacacatc aacgatctct
cgtaacgtaa ttgcgataag tcacttgttg 300gtatcacttg ggatcattct tgctgcagac
tatttcttga agcaggcgtt tgtagcagcg 360tctattaagt tcccaagtgc tttgtttggg
atgttctgta ttttctctgt tcttatgata 420tttgattcgg ttgttcctgc tgctgcaaat
ggtttgatga atttcttcga gcctgcgttt 480ctgtttatcc aaagatggct tcctttgttc
tatgttcctt ctcttgttgt tcttcctctt 540tctgttagag atattccggc tgcttcaggt
gtcaaaatct gctacattgt agccggtgga 600tggttggcgt cactttgtgt agcagggtac
acagctattg cagtgagaaa aatggtgaaa 660accgaaatga cggaagccga gcctatggca
aaaccatcac cattttcaac acttgagcta 720tggagttgga gtggaatctt tgttgtgtcg
tttgttggtg ctctgtttta ccctaattca 780ttggggacaa gtgcaagaac ttctctccct
ttccttcttt cttcaactgt gctaggttac 840attgtaggtt ctgggttgcc atcttctatt
aagaaagttt tccatccgat aatctgctgc 900gcgctatctg cagtacttgc tgctctagct
tttgggtatg cttcaggatc tggacttgat 960cctgttttag gaaactacct taccaaagta
gcatcagatc ctggtgctgg tgacatctta 1020atgggttttc ttggctctgt cattctctct
ttcgctttct ccatgttcaa acaaagaaag 1080ctcgtgaaga ggcacgcagc tgagatcttc
acatctgtga tagtttcaac ggtattctcg 1140ctctactcca ctgctcttgt tggacgttta
gtcggtttag aaccttcttt aacggtttca 1200atcctacctc gctgcatcac ggttgcattg
gcccttagca ttgtatcact ctttgaaggg 1260accaattcgt ctcttacagc agctgtagtc
gttgtgactg gtctgattgg agctaacttt 1320gtacaagttg ttcttgacaa actgcgttta
cgtgatccaa ttgctcgggg aattgcaact 1380gcttcaagtg ctcatggact tggaacagca
gctttgtcgg ctaaggagcc agaggctctt 1440cccttttgtg caatagctta tgctcttacc
ggaatcttcg gatcgttact gtgttctgtt 1500cctgccgtcc gacagagttt gctagcggtc
gtcggctag 153912936DNAZea mays 12atggctcgca
acgaggagaa ggcgcagtca atgctgaacc gcttcatcac gatgaagcag 60gaggagaagc
gcaagccccg agagcgccgg ccctacctcg cctccgagtg ccgcgacctc 120gccgacgccg
agcgctggcg ctctgagatc ctccgcgaga tcggcgccaa ggtcgccgag 180atccagaacg
agggtctcgg cgagcaccgc ctccgcgacc tcaacgacga gatcaacaaa 240ctcctccgcg
agcgcggcca ctgggagcgc cgcatcgtcg agctcggcgg ccgcgactac 300tcccgcagct
ccaacgcgcc gctcatgacc gacctcgacg gcaacatagt cgccgtcccc 360aacccctcgg
gtcgcggacc ggggtaccgc tactttggcg cggccaggaa gctccctggc 420gtgcgggagc
tcttcgacaa gccgcctgag atgcggaagc gacgcacccg ctacgagatc 480cacaagcgca
tcaacgccgg gtactacgga tactatgacg atgaggacgg cgtgctagag 540cgccttgagg
gccctgccgg gaagcgcatg cgggaggaga ttgtttcaga gtggcaccgt 600gtggaacggg
tgcggcggga ggccatgaag ggggtgatga gcggtgaggt ggctgcggct 660ggagggcgca
gcggggaggc tgctagagag gtgctgtttg agggggtgga ggaggaggtc 720gaagaggaga
ggaagcgtga ggaagagaag agggagaggg agaaaggcga ggaagttggg 780agggaattcg
ttgcacatgt gccgctacct gatgaaaagg agattgagcg catggtatta 840gagaggaaga
agaaggagct gcttagcaag tatgccagtg attccctgct ggttgagcag 900gaggaggcca
aggagatgct caatgtccga cgctag 936132049DNAZea
mays 13atggaccttg cacggagagg cggtgccgca ggcgcggacg acgaggggga gatcgagagg
60cacgagccgg cgcccgagga catggagtcc gaccccgcag cggcgcgcga gaaggagctg
120gagctggagc gggtgcagtc gtggcgggag caggtgactc tgcgcggcgt ggtggcggcg
180ctgctgatcg gcttcatgta cagcgtgatc gtgatgaaga tcgcgctcac cacggggctg
240gtgcccacgc tgaacgtctc cgcggcgctg atggcgttcc tggcgctccg cgggtggacg
300cgcgtgctgg agcgcctcgg cgtggcgcac cgccccttca cgcgccagga gaactgcgtc
360atcgagacct gcgccgtcgc gtgctacacc atcgcgttcg gcggtgggtt cggctccacg
420ctgctgggcc tggacaagaa gacgtacgag ctggccgggg cctcgccggc caacgttccg
480ggcagctaca aggaccctgg gttcggctgg atggccggat tcgtcgcggc gatcagcttc
540gccggcctcc taagcctgat ccccctcaga aaggttctgg tcattgacta caagctaact
600tacccaagcg ggactgcgac cgctgttctc ataaacgggt tccacaccaa gcaaggagac
660aagaacgcaa ggatgcaagt ccgagggttc ctcaagtact ttgggctcag cttcgtgtgg
720agctttttcc agtggttcta cacaggcggt gaagtttgcg gctttgttca gtttcctacg
780ttcggtctga aggcctggaa gcagacgttc ttctttgatt ttagcctcac gtacgttggt
840gcggggatga tctgttcgca cctcgtgaac atctccaccc tccttggtgc catcctgtca
900tgggggatac tgtggccact catcagcaag cagaaagggg agtggtaccc tgcgaacata
960cctgagagta gcatgaaaag cttatacggt tacaaggcct tcctctgcat agctctgatc
1020atgggagacg gtacatacca cttctttaaa gtcttcggtg tcactgttaa gagtctgcat
1080caacggctga gccgcaaacg tgctaccaac agagtggcaa acggtggaga cgaaatggcc
1140gcgcttgacg acctacagcg tgacgagatc ttcagcgacg ggtctttccc cgcctgggca
1200gcttacgccg ggtacgcggc gctgaccgtc gtctcagcgg tcatcatccc gcacatgttc
1260cggcaggtca agtggtacta cgtgatcgtg gcctacgtcc tcgcccctct cctcggcttc
1320gccaactcct acggcacggg gctcaccgac atcaacatgg cctacaacta cggcaagatc
1380gcgctcttca tcttcgcggc ctgggccggc agggacaacg gcgtcatcgc gggcctcgcc
1440ggcggcaccc tggtgaagca gctggtgatg gcgtccgcgg acctgatgca cgacttcaag
1500acgggccacc tgaccatgac gtcgcccagg tccctgctcg tggcgcagtt catcgggacg
1560gccatgggct gcgtcgtcgc gcccctcacg ttcctgctct tctacaacgc gttcgacatc
1620gggaacccca ccgggtactg gaaggcgccg tacggcctca tctaccgcaa catggcgatc
1680ctcggcgtgg agggcttctc cgtgctgccc aggcactgcc tcgcgctctc cgctgggttc
1740ttcgccttcg ccttcgtctt cagcgtcgcc cgggacgtcc tgccgcggaa gtacgccagg
1800ttcgtgcccc tgcccatggc catggccgtg ccgttcctcg tgggcgggag cttcgcgatc
1860gatatgtgcg tcgggagcct ggccgtcttt gtctgggaga aggtgaacag gaaggaggcc
1920gtgttcatgg tgcctgcggt tgcgtccggt ttgatctgtg gagacggcat atggaccttc
1980ccgtcttcca ttctcgctct ggccaagatc aagccaccga tttgcatgaa gttcactcct
2040ggaagctag
2049141362DNAArabidopsis thaliana 14atggctaaag tttattggcc ttatttcgat
cctgaatatg agaacttgag ctccagaatc 60aatcctccaa gtgtttctat agataacact
agctgcaaag aatgcactct tgtcaaggtg 120gacagtatga acaaacctgg aatactactt
gaagttgtgc aagtcctaac cgatctcgat 180ctcactatca ctaaagctta tatctcttct
gatggtggat ggttcatgga cgtattccat 240gtcaccgatc aacaaggaaa caaggttact
gatagcaaaa ccatcgatta catcgagaag 300gtgttaggac caaagggtca tgcttcggct
tcacaaaaca cttggcctgg taaaagagtc 360ggtgtccatt cattaggcga ccacacatcg
atagagatta ttgctcgtga tcgtcctggt 420ctcttgtcgg aggtttcagc cgtactagca
gacctcaaca ttaatgtggt ggcagctgaa 480gcatggactc acaaccgtag gattgcgtgt
gtcctctatg tgaatgacaa tgcaacttct 540agagccgttg atgatccaga aagattgtct
tccatggaag aacagcttaa caatgtgctg 600cgtgggtgcg aagaacaaga tgagaaattt
gctcggacga gtctctccat tgggtcgact 660cacgttgatc gaaggcttca tcagatgttt
ttcgctgata gagactacga agcagtgact 720aagcttgatg attctgcttc ttgcggattc
gagcccaaaa tcacggttga gcattgtgaa 780gagaaaggtt actccgtgat aaacgtgagc
tgcgaggatc gaccaaagct catgtttgac 840attgtatgca cgcttacgga tatgcaatac
attgtgtttc acgccacgat ttcatcaagc 900ggctctcatg cttctcagga gtatttcatc
agacacaaag acggttgcac tcttgacaca 960gaaggagaga aagagagagt tgtcaaatgt
ctggaagctg caatccatag acgagtcagc 1020gagggttgga gtttggagct ctgcgcaaag
gacagagttg gattactgtc ggaagtgaca 1080aggattctga gagagcacgg gctatcagtg
tcgagagctg gtgtgacaac agtaggagaa 1140caagccgtca acgttttcta tgtgaaagat
gcttcaggga atccagtgga tgtgaagacg 1200attgaggcgt tacgcggaga gattggacac
agtatgatga ttgacttcaa gaataaagtt 1260ccgagcagaa aatggaaaga agaaggtcaa
gccggaacag gaggaggatg ggccaaaacc 1320agtttcttct ttgggaattt gctggagaag
ttactgcctt ag 1362151005DNAEscherichia coli
15atggcgcaag ttagcagaat ctgcaatggt gtgcagaacc catctcttat ctccaatctc
60tcgaaatcca gtcaacgcaa atctccctta tcggtttctc tgaagacgca gcagcatcca
120cgagcttatc cgatttcgtc gtcgtgggga ttgaagaaga gtgggatgac gttaattggc
180tctgagcttc gtcctcttaa ggtcatgtct tctgtttcca cggcgtgcat gatgaatcca
240ttaattatca aactgggcgg cgtactgctg gatagtgaag aggcgctgga acgtctgttt
300agcgcactgg tgaattatcg tgagtcacat cagcgtccgc tggtgattgt gcacggcggc
360ggttgcgtgg tggatgagct gatgaaaggg ctgaatctgc cggtgaaaaa gaaaaacggc
420ctgcgggtga cgcctgctga tcagatagac attatcaccg gagcactggc gggaacggca
480aataaaaccc tgttggcatg ggcgaagaaa catcagattg cggccgtagg tttgtttctc
540ggtgacggcg acagcgtcaa agtgacccag cttgatgaag agttaggtca tgttggactg
600gcgcagccag gttcgcctaa gcttatcaac tccttgctgg agaacggtta tctgccggtg
660gtcagctcca ttggcgtaac agacgaaggg caactgatga acgtcaatgc cgaccaggcg
720gcaacggcgc tggcggcaac gctgggcgcg gatctgattt tgctctccga cgtcagcggc
780attctcgacg gcaaagggca acgcattgcc gaaatgaccg ccgcgaaagc agaacaactg
840attgagcagg gcattattac tgacggcatg atagtgaaag tgaacgcggc gctggatgcg
900gcccgcacgc tgggccgtcc ggtagatatc gcctcctggc gtcatgcgga gcagcttccg
960gcactgttta acggtatgcc gatgggtacg cggattttag cttag
1005161674DNAGlycine max 16atggcgctta aaaccctatc cactttcctc tcacctcttt
ctcttcccaa caccaaattc 60ccgcaattcc tcaccaccaa gccttccctc attctctgcg
agttccctcg ctctcagaaa 120tcgcgtttgc tcgccgccga ttcggaaggc accggcgccg
ccgctccttc tcccggcgag 180aagttcctcg aacgccagca gtccttcgaa gatgctaaga
tcattctcaa agaaaacaag 240aagaagagaa agaagaaaga caatgctata aaagcttcta
gagccgtcgc ttcttgctac 300ggctgcggcg ctccgttaca cacttccgat gccgatgccc
ctggctacgt tgatcccgaa 360acctatgaat tgaagaagaa acaccaccag cttcgaaccg
ttctgtgtag gcggtgccgg 420cttttgtctc atggcaagat gataactgcc gttggagggc
acggaggata tcctggcggt 480aaattattcg tcactgctga agagcttcga gaaaagttgt
ctcacctgcg tcacgagaaa 540gctctaatcg tcaaattggt tgatattgtt gacttcaatg
gcagtttttt gtctcgtgtg 600cgagatcttg ctggttctaa tccaataata ttggtggtga
ctaaggttga tctccttcct 660agagatactg atcttcattg tgttggggat tgggttgtag
aggctactat gagaaagaag 720ctaaatgttc tcagtgtcca tctgaccagt tccaaatcat
tggttggaat aactggagtg 780atatcggaaa tccagaaaga gaagaaggga cgagatgttt
acattctggg ttcagctaat 840gttgggaaat ctgctttcat caatgcttta ctaaaaacaa
tggctataaa tgatccagtg 900gctgcatctg cacaaagata caaaccaata caatctgcag
ttcctggaac taccttaggg 960ccaattcaaa ttaatgcttt cctaggagga gggaaattgt
acgacactcc tggagttcat 1020ctctaccata ggcaaactgc agttgttcat tctgaagatc
tacccatcct tgctccacaa 1080agccgactga ggggcctgtc tttcccaagt tctatattat
cttcagtaga ggaaggagct 1140tccaccatag tgaatggctt gaatgcattt tcaatatttt
ggggaggtct tgttagaatt 1200gatgtcttga aggttctccc agaaacttgt ttgacatttt
atggacccaa gagaatacca 1260attcatatgg tacccacaga gcaagcagtt gaattttatc
agacagaact tggagttctg 1320ctgaccccac caagtggagg agaaaatgct gagaactgga
aaggacttga atcagaacga 1380aaattgcaaa ttaaatttga agatgtggac agttatgatc
ccaaaccagc ttgtgatata 1440gctatatcag gtctaggatg gtttactgtt gagccagtta
gtcggtcact caaaatctca 1500caaccaaaac cggtagagac tgctggggaa ttgattttgg
ctgtgcacgt ccccaaggct 1560gttgagattt ttgtgaggtc accaatacca gtaggcaagg
ctggagcaga gtggtaccag 1620tatgtagaat taacagagaa acaagaggaa atgagaccaa
aatggtactt ttag 1674171281DNAZea mays 17atggcggcgg cgctcgcctc
ctcccgctac tgctggagcc gcccgtcgct gccgccccaa 60ccgacccgcg gccgccgctc
cgtcactagc tgcgcgctct ccggacgaga gaaaagaaac 120tcctttagct ggagagagtg
tgcaatttct gttgcattgt cagttggact aatcactggt 180gcaccaacgt ttggaccacc
ggcctatgct tcttctcttg aacctgttct tccagatgtg 240tctgttctta tctctggacc
tcccattaaa gatccaggtg ctttattgag atatgcttta 300ccaatagata ataaagctat
ccgtgaagtt caaaagccgc tggaggatat cactgacagc 360ctcaaggttg ctggtgttag
agccttggat tcagttgaaa gaaatgtcag acaagcatcg 420aaagcactga acaatgggag
aagcttaatt cttgctggcc ttgctgaacc aaaaagagca 480aatggagaag agttgttgaa
taagttggct gttggatttg aggagcttca aagaattgtg 540gaagacagaa atagggatgc
agtagctcca aagcagaaag agcttctcca gtatgttgga 600actgtagaag aagacatggt
cgatggcttt ccctttgaaa taccagaaga gtacagcaac 660atgcctcttc tcaaaggaag
agctactgtg gatatgaagg ttaagattaa ggacaatccc 720aacatggaag actgtgtatt
taggatagtt ctggatggat ataatgctcc tgtgactgct 780gggaacttcg tagatcttgt
caaacggaaa ttctatgatg gcatggaaat ccaaagagct 840gatggctttg ttgttcaaac
tggagatcca gaggggccag ctgagggctt tatcgatccc 900agcaccggca aaatccgtac
ggtacctctt gaaattatgg ttgatggtga taaggcgcct 960gtatatggtg aaacacttga
agaacttggt cgctacaagg ctcaaacaaa actccctttc 1020aacgcttttg gaacaatggc
tatggcaaga gaagaatttg atgacaattc tgcttctagc 1080caagtatttt ggctcttgaa
agagagtgag ctaacaccaa gcaatgccaa tatattggac 1140gggcggtacg cagtatttgg
atatgtaact gagaatgagg actacctggc tgacgtcaaa 1200gttggagatg tcatcgaatc
aatccaagtc gtctcaggct tggacaacct tgtcaaccca 1260agctacaaga ttgtaggata g
128118435DNAArabidopsis
thaliana 18atgatgcaag aattaggctt acaacgtttc tcaaacgacg tcgttcgctt
agacctcact 60cctccttctc aaacctcatc tacttctctt tccatcgacg aagaggaatc
aacggaagcc 120aagatccgac ggctgatatc ggagcatcct gtgatcatct tcagtagatc
ttcatgttgc 180atgtgccacg tcatgaagag actcttagca acgatcggcg taatccccac
cgtcatcgag 240ctcgatgatc acgaggtttc ctctcttccc acggctctac aagatgaata
ttccggtggc 300gtctccgtcg ttggtcctcc gccggcggtt ttcattggcc gtgagtgcgt
cggaggtctt 360gagtcccttg tcgctcttca cttaagtggt caacttgttc ctaagcttgt
ccaagttgga 420gctctttggg tatag
435191530DNAEscherichia coli 19atggcttcct ctatgctctc
ttccgctact atggttgcct ctccggctca ggccactatg 60gtcgctcctt tcaacggact
taagtcctcc gctgccttcc cagccacccg caaggctaac 120aacgacatta cttccatcac
aagcaacggc ggaagagtta actgcatgca ggtgtggcct 180ccgattggaa agaagaagtt
tgagactctc tcttaccttc ctgaccttac cgattccggt 240ggtcgcgtca actgcatgca
ggccatgagc aacaatgaat tccatcagcg tcgtctttct 300gccactccgc gcggggttgg
cgtgatgtgt aacttcttcg cccagtcggc tgaaaacgcc 360acgctgaagg atgttgaggg
caacgagtac atcgatttcg ccgcaggcat tgcggtgctg 420aataccggac atcgccaccc
tgatctggtc gcggcggtgg agcagcaact gcaacagttt 480acccacaccg cgtatcagat
tgtgccgtat gaaagctacg tcaccctggc ggagaaaatc 540aacgcccttg ccccggtgag
cgggcaggcc aaaaccgcgt tcttcaccac cggtgcggaa 600gcggtggaaa acgcggtgaa
aattgctcgc gcccataccg gacgccctgg cgtgattgcg 660tttagcggcg gctttcacgg
tcgtacgtat atgaccatgg cgctgaccgg aaaagttgcg 720ccgtacaaaa tcggcttcgg
cccgttccct ggttcggtgt atcacgtacc ttatccgtca 780gatttacacg gcatttcaac
acaggactcc ctcgacgcca tcgaacgctt gtttaaatca 840gacatcgaag cgaagcaggt
ggcggcgatt attttcgaac cggtgcaggg cgagggcggt 900ttcaacgttg cgccaaaaga
gctggttgcc gctattcgcc gcctgtgcga cgagcacggt 960attgtgatga ttgctgatga
agtgcaaagc ggctttgcgc gtaccggtaa gctgtttgcc 1020atggatcatt acgccgataa
gccggattta atgacgatgg cgaaaagcct cgcgggcggg 1080atgccgcttt cgggcgtggt
cggtaacgcg aatattatgg acgcacccgc gccgggcggg 1140cttggcggca cctacgccgg
taacccgctg gcggtggctg ccgcgcacgc ggtgctcaac 1200attatcgaca aagaatcact
ctgcgaacgc gcgaatcaac tgggccagcg tctcaaaaac 1260acgttgattg atgccaaaga
aagcgttccg gccattgctg cggtacgcgg cctggggtcg 1320atgattgcgg tagagtttaa
cgatccgcaa acgggcgagc cgtcagcggc gattgcacag 1380aaaatccagc aacgcgcgct
ggcgcagggg ctgctcctgc tgacctgtgg cgcatacggc 1440aacgtgattc gcttcctgta
tccgctgacc atcccggatg cgcaattcga tgcggcaatg 1500aaaattttgc aggatgcgct
gagcgattag 1530201473DNASynechocystis
sp. 20atggcttcct ctatgctctc ttccgctact atggttgcct ctccggctca ggccactatg
60gtcgctcctt tcaacggact taagtcctcc gctgccttcc cagccacccg caaggctaac
120aacgacatta cttccatcac aagcaacggc ggaagagtta actgcatgca ggtgtggcct
180ccgattggaa agaagaagtt tgagactctc tcttaccttc ctgaccttac cgattccggt
240ggtcgcgtca actgcatgca ggccatgacc ccagaattga atcctaattt tcccgaagaa
300actacctccg atgcttggct gaccccagca gatgccggcc aggatggtga tgcccaggaa
360ccggcggaag atgggggaga agaaggagta gtgtcggaag aactggccct gcctgaggac
420ttacctccta tggatgccat ggtggcggca gtggaagaaa tgactccggt ggtggtgccc
480gaaactgtac cagaaacaga aaccccagcc ttagaggatt tggtcgccca aaagaccgcc
540ctggaaaagg acattgccgc tctgcaacgg gaaaaagccc agtggtatgg ccagcagttc
600cagcaattac agcgggaaat ggcccggtta gtggaggaag gcaccaggga attagggcaa
660agaaaagcag ctctggaaaa ggaaattgag aagttagagc gccgtcagga acggattcaa
720caggaaatgc gtaccacttt tgccggggct tcccaggagt tggccatccg cgtgcagggc
780tttaaggatt atttggtggg gagtttgcag gatttggttt ccgccgccga ccagttggaa
840ttaggggtgg gggacagttg ggagtcttcc tctacccatg gggatgcgat tattgaaaat
900gccgacccaa ctccggtggt gagttttgcg gagcagggtt ttagtagcca aaaacgacaa
960atccaagctt tgctggagca ataccgcact cgccctgatt attacggtcc cccttggcag
1020ttgcgtcgta cctttgagcc agtccacgcc gaacggattg agaattggtt ctttaccctg
1080ggcggtcggg gagcaatcct cagtttagac agtcgtttac aaaatatttt ggtgggttca
1140gcggcgatcg ccattttgaa tcagctctac ggcgatcgtt gtcgggcgtt aattttggcg
1200gccaccccag aaagattggg ggaatggcga cggggtttac aggattgttt gggtatttcc
1260cgcagtgact ttggcccaga ccggggcatt gttttgtttg aatcggccaa tgccttgatc
1320cagcgggcgg aaagattggt cggcgatcgc caaatgccgt tggtgttggt ggatgaaaca
1380gaggaacaaa ttgacttagc cctgttgcaa ttcccccttt tactggcctt tgcacctagt
1440taccaagtcg gaggcagtaa ctatttttct tag
1473211215DNAZea mays 21atggccggcg aactgcgcca ccgccgcgcg ccgtcggagg
acgagggcgt cgcctcctct 60caaagactcg actccgcccc cgcaggcaac ggcaaggctg
gcacttcgtc cggcggcggc 120gagggggcgg agccgcgggg cgggaagagg gacgcgctag
ggtggctgga gtggtgccgc 180ggttggatgg ccatcgtggg ggagttcctc ttccagcgca
tcgccgccag ccacctggcc 240aacccgctcg agttgccccc gctcgatggc gtctccatcg
tcgtcaccgg cgccaccagc 300ggcatcgggc tcgagatcgc aaggcaactc gctctcgctg
gggcacatgt tgttatggct 360gtaaggagac ccaaggtggc acaagagttg attcagaagt
ggcagaatga aaattcagaa 420acaggaagac cactaaatgc cgaggtgatg gaacttgacc
tgctctccct cgactcggtc 480gtaaaatttg ctgatgcttg gaatgctcgt atggcaccgc
tgcacgtgtt gatcaacaat 540gctggcatct tcgctatagg agaaccccaa catttttcga
aggatggaca tgaagaacac 600atgcaagtga accatcttgc acctgcatta ctggcgatgc
tgcttatacc ttcccttctc 660cgaggttctc ccagcagaat cgttaacgtt aattcaatca
tgcacagtgt aggttttgtt 720gatgctgaag atttcaactt gagaaaacat aaatatagaa
gttggttggc gtattcaaat 780agcaagttgg cacaggtaaa atttagtagc atgcttcata
agagaattcc tgcagaagct 840ggcatcagca taatttgtgc ttctcctgga attgtcgaca
cgaatgttac aagagacctt 900cctaagattg ttgtagctgc ataccgtttt cttccctact
tcatattcga tggtcaagaa 960ggttctagga gtgcactgtt tgcggcatgt gacccccaag
ttccagagta ctgtgagatg 1020ctcaagtcgg aagactggcc agtctgtgct tgcattaact
acgactgtaa tccgatgaac 1080gcgtctgaag aagcgcacag ccttgaaacc tcgcagctgg
tctgggagaa gacgctcgag 1140atgatcggcc ttccgccgga tgccctggac aagctcatcg
ccggagaaac agtgccgtgc 1200cgttatggac aatag
1215222829DNAZea mays 22atggagggcg acgacttcac
gccggagggc ggcaagctcc ccgagttcaa gctagatgcg 60agacaagcgc agggtttcat
ctccttcttc aagaagctgc cgcaggatcc ccgggccgtt 120cgtctcttcg atcgcaggga
ttattacact gcccatggcg agaatgctac gtttatcgca 180aggacatact accacacaat
gtctgcctta cgtcaactag gtagcacctc tgatggaatc 240ttaagtgcca gcgtgagcaa
ggctatgttt gagaccattg cccgcaacat tttgttggaa 300aggactgact gtacattgga
actctatgag ggaagtgggt caaattggag gttaacaaag 360tccggaacac ctggaaatat
tggtagtttt gaagacattc tgtttgcaaa caatgacatg 420gaagattcac cagtgattgt
tgctttgttt ccagcgtgcc gggaaagtca gctgtatgta 480gggcttagtt ttttggatat
gaccaatagg aagcttgggt tggctgagtt tcccgaagat 540agccgattca ctaatgttga
atcagctctt gttgcattag gttgcaagga gtgtcttctc 600ccagcagatt gtgaaaaatc
cattgaccta aatccccttc aagacgtcat tagtaactgt 660aatgttctgt tgactgagaa
aaagaaggct gacttcaaat ccagggatct cgcacaagat 720cttggtagaa taatcagggg
ttctgttgag cctgtacgtg atctactatc tcagtttgac 780tatgctcttg gtccccttgg
agctctttta tcttatgccg agttgctagc agatgacact 840aactatggaa attacacaat
tgagaagtac aatttgaact gctacatgcg acttgattct 900gctgcagttc gagcattaaa
cattgcagaa gggaaaactg atgtaaacaa gaacttcagt 960ttgtttggtt tgatgaacag
aacttgtact gttgggatgg gaaaaagatt gctgaacaga 1020tggctgaaac aacctctatt
agatgttaat gaaattaata accgactaga catggttcag 1080gcttttgtag aagacccaga
acttcgtcag ggactccggc aacaacttaa aaggatatca 1140gatattgatc gtctaacaca
tagtctccga aagaaatcag ctaatctgca gcctgttgtt 1200aagctttatc agtcctgtag
cagaatccca tacatcaagg gcattcttca gcaatataat 1260ggccaatttt caacattgat
aaggtcaaag tttcttgaac cgttagaaga atggatggca 1320aagaatcgat ttggtcgttt
ttcttctctt gttgagacag ctattgatct tgctcagctg 1380gagaatggag agtacagaat
atctcctcta tattcttctg acttgggtgt actaaaggat 1440gagctttctg tggttgaaaa
ccacataaac aatctgcacg tggatacagc tagtgatctg 1500gatctttctg ttgataagca
actgaagcta gaaaaaggat cccttggaca tgtgttcaga 1560atgtcaaaga aagaggaaca
gaaagtcagg aagaaactca ctggcagcta cttaatcata 1620gaaactcgta aagatggtgt
aaagttcaca aattctaagc tgaaaaatct aagtgatcaa 1680taccaggcat tgtttggtga
gtacacaagt tgtcagaaaa aggtggttgg tgatgtagtg 1740agggtttcag gcacattctc
agaggtattt gaaaattttg ctgcagttct gtcggagttg 1800gatgttttac aaagttttgc
tgatttggca actagttgcc cagttcctta tgttaggcca 1860gacatcactg cgtcggatga
aggagatatt gttctactgg gtagcagaca tccttgtcta 1920gaggcacaag atggtgttaa
ctttataccc aatgattgca ctctggtgag agggaaaagt 1980tggtttcaga tcatcactgg
accaaacatg ggaggaaaat ccacatttat aagacaggtt 2040ggtgtaaatg tattgatggc
acaagttggt tcctttgtac cttgtgatca agcaagtatt 2100agtgtgaggg attgtatttt
tgctcgtgtt ggcgctggtg attgccaact tcatggtgta 2160tcaactttta tgcaagaaat
gcttgaaaca gcatccatcc taaaaggcgc ctctgataag 2220tctcttataa ttattgatga
gctggggcgt ggaacttcca catatgatgg atttggtctt 2280gcatgggcta tctgtgagca
tcttatggaa gtgactcgag cgcctacctt gtttgcaacc 2340catttccatg aactaactgc
attagcacat agaaatgatg atgagcacca acacatttca 2400gacatcggag ttgcaaatta
tcacgtgggt gctcacatag acccattaag taggaagtta 2460actatgcttt acaaggttga
acctggtgca tgcgaccaaa gttttggtat tcatgttgca 2520gaatttgcta attttccaga
agctgttgtt gcccttgcga aaagcaaagc agcagagtta 2580gaagactttt ctactacacc
taccttttcc gatgatttga aagacgaggt tggatcaaag 2640cgcaagaggg tatttagccc
agatgacatc accagaggag ctgcacgggc tcggcttttc 2700cttgaggaat tcgccgcatt
gcctatggat gagatggatg ggagcaagat attggagatg 2760gccaccaaga tgaaagctga
cttgcagaaa gatgcagctg acaatccttg gctccagcag 2820ttcttctag
2829231035DNAArabidopsis
thaliana 23atggagtgga ttcgaggaga aactatcgga tacggaactt tttctacagt
aagtctagcg 60acgcggtcta ataacgattc cggcgagttt cctccgttaa tggctgtgaa
atctgcagac 120tcatacggcg ctgcttctct ggcaaacgag aaatcagttc tagataatct
cggagacgat 180tgcaacgaga tcgtacggtg tttcggcgag gatcggacgg tcgaaaacgg
tgaagagatg 240cataatttgt tcttggaata cgcttctaga ggaagcttag agagttatct
taagaaatta 300gccggtgaag gtgtaccgga atccaccgtg cgtcgccaca caggatcggt
gcttagaggt 360ctacgacaca tccacgctaa cggattcgct cactgtgatt taaaactcgg
gaatattctg 420ttgttcggtg acggcgccgt taagattgcg gattttggat tggcgaagag
aattggggat 480ttaacggcgt taaattacgg tgtgcagatt agaggtacgc cgttgtacat
ggcgccggaa 540tctgttaacg ataacgagta cggatcagaa ggtgacgtgt gggctttagg
atgcgtagta 600gttgagatgt ttagtggtaa aacggcatgg agtttaaaag aagggtcgaa
cttcatgtcg 660ttgttgttac gcatcggtgt tggtgacgag gttccgatga ttcccgagga
gttgtcggaa 720caaggaagag attttttgtc aaagtgtttc gttaaagatc ccaaaaagag
atggacggct 780gagatgcttc taaaccatcc atttgtaacc gtcgatgttg atcacgacgt
tttagtcaaa 840gaagaagatt tcgttgttaa tatgaaaaca gaggacgtct cgacatcgcc
gagatgccca 900ttcgaatttc ccgattgggt ttcggtttct tccggttcac aaacgatcga
ttcgccggat 960gagagagttg ctagtttggt gactgatatg atccctgatt ggtctgttac
caatagctgg 1020gtcaccgtac ggtga
1035243564DNAArabidopsis thaliana 24atgagaaacc attgcttaga
actctcttcc aattgttcct ccattttcgc ttcttccaaa 60tccaatcctc gtttctctcc
ttccaagctc tcctattcca ctttcttctc tcgctctgcc 120atctattaca gatcaaaacc
aaaacaagcc tcgtcttctt cttccttctc cactttcccc 180ccatgtctca atcggaaaag
ctccctcacg catgttctca aacccgtctc agagctcgcc 240gacaccacta ccaagccttt
ttctccggag atcgtcggca agagaaccga tctgaagaag 300attatgattc tcggcgctgg
tccgattgtc attggacaag cttgtgagtt tgattactct 360ggtactcaag cttgtaaagc
cttaagagaa gagggctatg aggttatcct gatcaattcg 420aatcctgcca ctatcatgac
tgatccggaa actgctaatc ggacttatat cgctccgatg 480actcctgagc ttgtcgagca
ggttattgag aaagagaggc ctgacgcttt gttaccaacc 540atgggtggtc aaaccgcatt
gaacctcgcg gttgctcttg ctgagagtgg tgctttggag 600aaatacggtg ttgaattgat
aggagctaag cttggtgcga ttaagaaagc tgaagatcgt 660gagttgttca aggatgcgat
gaagaacatt gggctaaaga ctccaccttc agggattggg 720accactcttg atgagtgttt
tgacattgct gagaaaattg gtgagttccc tttgattatc 780cgtcctgcgt ttactttagg
tggtactggt ggtggaattg cgtataacaa agaggagttt 840gagtctatat gtaaatcggg
tttggctgcg agtgcgacaa gtcaagttct tgtggagaaa 900tccttgttgg gttggaaaga
atatgagctt gaggtgatga gagacttagc tgacaatgtt 960gtcattatct gttccattga
gaatattgat cctatgggtg tgcacactgg tgattccatc 1020actgtggcac ctgcacagac
tctaacggat agagagtacc agcggcttag ggattattcc 1080attgcgatta tacgggagat
tggtgttgag tgtggtggat ctaatgtgca gtttgctgtc 1140aacccggttg atggtgaagt
tatgatcata gagatgaacc ctagggtctc aagatcttct 1200gctcttgctt ccaaggctac
agggtttccc attgctaaaa tggctgccaa gttgtctgtt 1260ggctatacct tggatcagat
tcctaatgat atcacgagga aaacaccggc tagcttcgag 1320ccctccatcg attatgtggt
gactaagatt cctcgatttg catttgaaaa gtttccagga 1380tctcagccat tgctaacgac
ccagatgaaa tctgttgggg aatctatggc tctcggccgt 1440acattccaag aatctttcca
gaaagctctg aggtctctgg agtgtggatt ctcgggttgg 1500ggttgtgcaa aaattaaaga
gctagattgg gactgggatc agctgaaata cagcctaaga 1560gtcccaaatc ctgacaggat
ccatgcgata tatgctgcca tgaaaaaggg tatgaaaatt 1620gatgaaatct acgagttgag
catggtggac aagtggttcc taacccagct taaagagctc 1680gtggacgtcg aacagtatct
tatgtccgga accttgtcag agattacaaa agaagacctt 1740tacgaagtca aaaagcgggg
atttagtgac aagcaaatcg cttttgctac aaagacaacc 1800gaggaagaag tccgtaccaa
gcggatttct ctaggagttg ttccatctta caagagagtg 1860gatacatgtg ctgcagagtt
cgaagcgcat acaccataca tgtactcttc atatgatgtt 1920gaatgtgaat cagctccaaa
caacaagaag aaggttttga ttttgggtgg agggccaaac 1980cgcattggtc aagggattga
atttgattac tgttgttgcc acacatcttt cgccttacag 2040gatgctggat atgagaccat
aatgttgaac tcaaatcctg aaacagtatc cacagattat 2100gatacaagcg ataggctcta
ttttgaacct ctcacaatcg aggatgttct caatgttatc 2160gaccttgaga aacctgatgg
cataatagtg caatttggtg gtcaaactcc tctgaaactt 2220gctctgccga tcaaacatta
tttggataag cacatgccca tgagcttgag cggagcggga 2280cctgttcgca tctggggtac
atcacctgac tccattgacg ctgctgaaga cagagagagg 2340ttcaatgcaa ttctcgacga
gctgaagatt gagcagccca agggaggcat tgcaaagagc 2400gaagctgatg cattagccat
agcaaaggag gtagggtacc cagttgtggt aagaccttct 2460tatgttctag gtggacgagc
aatggagatc gtttatgatg acagtagact aataacctat 2520ttggaaaatg cggtacaagt
tgacccagag agacctgttt tggtagataa atatctttct 2580gatgccattg agatcgacgt
tgataccctt actgattcct atggaaatgt ggtgattggt 2640ggaataatgg agcatatcga
acaagctggt gtgcattctg gtgactcagc ttgtatgctt 2700ccaacacaaa ccatcccagc
ttcttgtttg caaactattc gaacatggac cactaagctg 2760gcgaagaagc taaatgtatg
tgggctgatg aactgtcagt acgcaatcac aacatctggg 2820gatgtttttt tgctggaagc
caatccccga gcttcccgta ctgtcccttt tgtgtcaaaa 2880gccattggac accctcttgc
caagtatgca gcgctggtca tgtcgggcaa atctctcaaa 2940gatcttaact ttgaaaaaga
agttatccct aaacatgtct ctgtgaaaga agctgttttc 3000ccgtttgaga agttccaagg
atgcgatgtg atactcgggc cagagatgag aagcacagga 3060gaagtgatga gcatcagttc
tgaattctca agtgcgtttg caatggctca gatcgctgca 3120ggtcaaaagc tacctctatc
aggcacagtc ttcctcagct taaacgatat gaccaaaccg 3180cacctggaga aaatcgcggt
gtccttcctc gagcttgggt tcaaaatagt tgccacctcg 3240ggaacagctc atttcctgga
actgaaaggc attccagtgg agagagtgtt gaagttgcat 3300gaaggaagac cacatgctgc
tgatatggtg gcgaatggtc agatccattt gatgttgatc 3360acaagctcgg gtgatgctct
tgatcagaaa gatgggagac agctcagaca aatggctcta 3420gcatacaagg tacctgttat
aaccactgtt gctggtgcat tggccactgc tgagggaatc 3480aagagcttga agtcaagtgc
cattaaaatg accgctcttc aggacttctt tgaggtaaag 3540aatgtatctt ctttgctcgt
ctga 356425825DNAGlycine max
25atgagagcca aattgtttgt gttcccaata cgaggcagga actggtgctt ctccagaacc
60atcgatcact ctctctccgc ttcccatgct tcctctcaat ccccctcaac cctcaaagac
120ttgtggacca acatcaacgt tggtgataaa cccctgaaca ccaaaactga gctctttgtc
180gattacatcg ccaacaagat gaataatgct tggattggct tggagaaggc gccggagggg
240tctttcaaga acaagattca tgggttgggg ttgcggctct tgtcgcgggt taagccctct
300gagatatttt tgaagtctat atcgaaggaa atcactagtg ttgaaatcat ttatccatca
360agtttgaatg ctcaacttgt tcgtcgaaga ctaagacaca ttgctgtgag gggagcagtt
420atccatcgga attacttata cggtttagtt tcgttgattc cattgacttc agcacttagc
480attttacctt tgcctaatgt tccgttcttc tgggttttat ttcgcactta ttctcattgg
540agagccttgc agggaagtga gaggctgttt caactagtct cagataacag caagacttca
600aacacttgta catatgaaaa gaaaactgag cacaaggaat ctaaaagtca aagacatagt
660tcaaatgaac cttgttgggt gttgaggcca tccaaagaac ttgagaatct tgtccatcta
720gaagatggtc aagagagtct tagtcaacat gccatcataa acatttgcaa gatctatgac
780ttgaacccag tagatgttat aaaatacgag aagtccgtct tttaa
82526621DNAArabidopsis thaliana 26atggcgaaag agtccaccac catcgacgtc
ggcgagccaa gcactgttac caaaagttca 60agccatgtcg taaaggacgc gaagaagaag
ggctttgtgg cagtcgcctc aagaggtggt 120gccaagagag gtttggctat attcgatttc
ctcctccgtt tggcggccat agcagtcact 180attggggctg cctctgtcat gtacaccgcc
gaggaaactc ttcccttctt tactcagttc 240ctccagttcc aagccggtta cgatgacctt
cctgcgtttc agtactttgt gatagccgta 300gccgtagtcg ctagctatct cgtcctttca
cttccattct ccatcgtatc cattgtccgt 360ccacatgctg tcgcgccccg gctgatcctc
ctcatttgcg atactctggt cgtgacgctc 420aacacatcag cagcagcagc ggcagcatca
atcacctacc ttgcacacaa cggcaaccaa 480agcaccaact ggctccctat ctgtcagcag
tttggagact tctgccagaa cgttagcacc 540gcggttgtgg ctgattctat cgcgattctc
ttcttcatcg ttcttatcat catctcagcc 600atcgccctca agaggcattg a
621271236DNAArtificialcodon optimized
Escherichia coli Asparagine synthetase A (AsnA) gene 27atggcaacag
caacatcagc ttctctgttt tcaactgttt cttcatctta ctccaaagct 60agctccatac
cacattcaag actccaatct gtgaaattca actcagtccc tagcttcacc 120ggtctcaaat
caacctctct catctccgga tctgattcct cttccttagc caagactcta 180cgcggttccg
taacgaaagc acaaacatct gacaagaagc cttacggatt caaaatcaac 240gctatgaaga
ccgcctacat cgctaagcag cgccagatct ccttcgtgaa gtcccacttc 300tctaggcagc
tagaggagcg cctaggcctg atcgaggtgc aggcccccat ccttagccgc 360gtgggtgacg
gcacccagga caaccttagc ggctgcgaga aggccgtgca ggtgaaggtg 420aaggccctcc
ccgacgccca gttcgaggtg gtccactccc tcgccaagtg gaagcgccag 480accctcggcc
agcacgactt cagcgccggc gagggcctct acacccacat gaaggccctc 540cgccccgacg
aggacaggct ctcccccctc cactccgtgt acgtggacca gtgggactgg 600gagagggtca
tgggcgacgg cgagaggcag ttctccaccc tcaagagcac tgtcgaggcc 660atctgggccg
gcatcaaggc tactgaggct gcggtcagcg aggaattcgg cctcgctcct 720ttcctccctg
accagatcca ctttgtccac tctcaggagc tcctgtctag gtaccctgac 780ctcgacgcta
agggccggga gcgggctatc gctaaggacc tcggtgctgt ctttctggtc 840ggtatcggtg
ggaaactgtc tgacggtcac cggcacgatg tccgtgcgcc tgattatgat 900gattggtcga
ctccgtcgga gctgggtcat gcgggtctga acggggatat tctggtttgg 960aatccggttc
tggaggatgc gtttgagctg tcgtcaatgg ggattcgtgt tgatgcggat 1020acgctgaaac
atcagctggc actgacgggg gatgaggata gacttgaact tgaatggcat 1080caggcattac
ttcgtgggga aatgccgcag acaattgggg gaggaattgg acaatcaaga 1140cttacaatgc
ttttgcttca attgccacat ataggacaag ttcaatgcgg agtttggcca 1200gcagcagttc
gagaaagtgt accaagtttg ttgtga
12362854PRTArabidopsis thaliana 28Met Val Asn Asn Val Val Ser Ile Glu Lys
Met Lys Ala Leu Trp His1 5 10
15Ser Glu Val His Asp Glu Gln Lys Trp Ala Val Asn Met Lys Leu Leu
20 25 30Arg Ala Leu Gly Met Phe
Ala Gly Gly Val Val Leu Met Arg Ser Tyr 35 40
45Gly Asp Leu Met Gly Val 5029315PRTArabidopsis thaliana
29Met Val Gly Trp Ala Ile Ala Leu His Gly Gly Ala Gly Asp Ile Pro1
5 10 15Ile Asp Leu Pro Asp Glu
Arg Arg Ile Pro Arg Glu Ser Ala Leu Arg 20 25
30His Cys Leu Asp Leu Gly Ile Ser Ala Leu Lys Ser Gly
Lys Pro Pro 35 40 45Leu Asp Val
Ala Glu Leu Val Val Arg Glu Leu Glu Asn His Pro Asp 50
55 60Phe Asn Ala Gly Lys Gly Ser Val Leu Thr Ala Gln
Gly Thr Val Glu65 70 75
80Met Glu Ala Ser Ile Met Asp Gly Lys Thr Lys Arg Cys Gly Ala Val
85 90 95Ser Gly Leu Thr Thr Val
Val Asn Pro Ile Ser Leu Ala Arg Leu Val 100
105 110Met Glu Lys Thr Pro His Ile Tyr Leu Ala Phe Asp
Ala Ala Glu Ala 115 120 125Phe Ala
Arg Ala His Gly Val Glu Thr Val Asp Ser Ser His Phe Ile 130
135 140Thr Pro Glu Asn Ile Ala Arg Leu Lys Gln Ala
Lys Glu Phe Asn Arg145 150 155
160Val Gln Leu Asp Tyr Thr Val Pro Ser Pro Lys Val Pro Asp Asn Cys
165 170 175Gly Asp Ser Gln
Ile Gly Thr Val Gly Cys Val Ala Val Asp Ser Ala 180
185 190Gly Asn Leu Ala Ser Ala Thr Ser Thr Gly Gly
Tyr Val Asn Lys Met 195 200 205Val
Gly Arg Ile Gly Asp Thr Pro Val Ile Gly Ala Gly Thr Tyr Ala 210
215 220Asn His Leu Cys Ala Ile Ser Ala Thr Gly
Lys Gly Glu Asp Ile Ile225 230 235
240Arg Gly Thr Val Ala Arg Asp Val Ala Ala Leu Met Glu Tyr Lys
Gly 245 250 255Leu Ser Leu
Thr Glu Ala Ala Ala Tyr Val Val Asp Gln Ser Val Pro 260
265 270Arg Gly Ser Cys Gly Leu Val Ala Val Ser
Ala Asn Gly Glu Val Thr 275 280
285Met Pro Phe Asn Thr Thr Gly Met Phe Arg Ala Cys Ala Ser Glu Asp 290
295 300Gly Tyr Ser Glu Ile Ala Ile Trp
Pro Asn Asn305 310 31530726PRTArabidopsis
thaliana 30Met Pro Ser His Pro Asn Phe Ile Phe Arg Trp Ile Gly Leu Phe
Ser1 5 10 15Asp Lys Phe
Arg Arg Gln Thr Thr Gly Ile Asp Glu Asn Ser Asn Leu 20
25 30Gln Ile Asn Gly Gly Asp Ser Ser Ser Ser
Gly Ser Asp Glu Thr Pro 35 40
45Val Leu Ser Ser Val Glu Cys Tyr Ala Cys Thr Gln Val Gly Val Pro 50
55 60Ala Phe His Ser Thr Ser Cys Asp Gln
Ala His Ala Pro Glu Trp Arg65 70 75
80Ala Ser Ala Gly Ser Ser Leu Val Pro Ile Gln Glu Gly Ser
Val Pro 85 90 95Asn Pro
Ala Arg Thr Arg Phe Arg Arg Leu Lys Gly Pro Phe Gly Glu 100
105 110Val Leu Asp Pro Arg Ser Lys Arg Val
Gln Arg Trp Asn Arg Ala Leu 115 120
125Leu Leu Ala Arg Gly Met Ala Leu Ala Val Asp Pro Leu Phe Phe Tyr
130 135 140Ala Leu Ser Ile Gly Arg Thr
Thr Gly Pro Ala Cys Leu Tyr Met Asp145 150
155 160Gly Ala Phe Ala Ala Val Val Thr Val Leu Arg Thr
Cys Leu Asp Ala 165 170
175Val His Leu Trp His Val Trp Leu Gln Phe Arg Leu Ala Tyr Val Ser
180 185 190Arg Glu Ser Leu Val Val
Gly Cys Gly Lys Leu Val Trp Asp Pro Arg 195 200
205Ala Ile Ala Ser His Tyr Ala Arg Ser Leu Thr Gly Phe Trp
Phe Asp 210 215 220Val Ile Val Ile Leu
Pro Val Pro Gln Ala Val Phe Trp Leu Val Val225 230
235 240Pro Lys Leu Ile Arg Glu Glu Lys Val Lys
Leu Ile Met Thr Ile Leu 245 250
255Leu Leu Ile Phe Leu Phe Gln Phe Leu Pro Lys Ile Tyr His Cys Ile
260 265 270Cys Leu Met Arg Arg
Met Gln Lys Val Thr Gly Tyr Ile Phe Gly Thr 275
280 285Ile Trp Trp Gly Phe Ala Leu Asn Leu Ile Ala Tyr
Phe Ile Ala Ser 290 295 300His Val Ala
Gly Gly Cys Trp Tyr Val Leu Ala Ile Gln Arg Val Ala305
310 315 320Ser Cys Ile Arg Gln Gln Cys
Met Arg Thr Gly Asn Cys Asn Leu Ser 325
330 335Leu Ala Cys Lys Glu Glu Val Cys Tyr Gln Phe Val
Ser Pro Thr Ser 340 345 350Thr
Val Gly Tyr Pro Cys Leu Ser Gly Asn Leu Thr Ser Val Val Asn 355
360 365Lys Pro Met Cys Leu Asp Ser Asn Gly
Pro Phe Arg Tyr Gly Ile Tyr 370 375
380Arg Trp Ala Leu Pro Val Ile Ser Ser Asn Ser Leu Ala Val Lys Ile385
390 395 400Leu Tyr Pro Ile
Phe Trp Gly Leu Met Thr Leu Ser Thr Phe Ala Asn 405
410 415Asp Leu Glu Pro Thr Ser Asn Trp Leu Glu
Val Ile Phe Ser Ile Val 420 425
430Met Val Leu Ser Gly Leu Leu Leu Phe Thr Leu Leu Ile Gly Asn Ile
435 440 445Gln Val Phe Leu His Ala Val
Met Ala Lys Lys Arg Lys Met Gln Ile 450 455
460Arg Cys Arg Asp Met Glu Trp Trp Met Lys Arg Arg Gln Leu Pro
Ser465 470 475 480Arg Leu
Arg Gln Arg Val Arg Arg Phe Glu Arg Gln Arg Trp Asn Ala
485 490 495Leu Gly Gly Glu Asp Glu Leu
Glu Leu Ile His Asp Leu Pro Pro Gly 500 505
510Leu Arg Arg Asp Ile Lys Arg Tyr Leu Cys Phe Asp Leu Ile
Asn Lys 515 520 525Val Pro Leu Phe
Arg Gly Met Asp Asp Leu Ile Leu Asp Asn Ile Cys 530
535 540Asp Arg Ala Lys Pro Arg Val Phe Ser Lys Asp Glu
Lys Ile Ile Arg545 550 555
560Glu Gly Asp Pro Val Gln Arg Met Ile Phe Ile Met Arg Gly Arg Val
565 570 575Lys Arg Ile Gln Ser
Leu Ser Lys Gly Val Leu Ala Thr Ser Thr Leu 580
585 590Glu Pro Gly Gly Tyr Leu Gly Asp Glu Leu Leu Ser
Trp Cys Leu Arg 595 600 605Arg Pro
Phe Leu Asp Arg Leu Pro Pro Ser Ser Ala Thr Phe Val Cys 610
615 620Leu Glu Asn Ile Glu Ala Phe Ser Leu Gly Ser
Glu Asp Leu Arg Tyr625 630 635
640Ile Thr Asp His Phe Arg Tyr Lys Phe Ala Asn Glu Arg Leu Lys Arg
645 650 655Thr Ala Arg Tyr
Tyr Ser Ser Asn Trp Arg Thr Trp Ala Ala Val Asn 660
665 670Ile Gln Met Ala Trp Arg Arg Arg Arg Lys Arg
Thr Arg Gly Glu Asn 675 680 685Ile
Gly Gly Ser Met Ser Pro Val Ser Glu Asn Ser Ile Glu Gly Asn 690
695 700Ser Glu Arg Arg Leu Leu Gln Tyr Ala Ala
Met Phe Met Ser Ile Arg705 710 715
720Pro His Asp His Leu Glu 72531467PRTArabidopsis
thaliana 31Met Ile Leu Asp Leu Gly Phe Pro Cys Phe Val Pro Pro Arg Thr
Ser1 5 10 15Ser Arg Glu
Asp Asn Lys Ala Trp Leu Leu Ala Glu Thr Glu Pro Lys 20
25 30Leu Ile Asp Ser Glu Gln His Ser Leu Gln
Ser Ser Phe Arg Phe Ser 35 40
45Leu Cys Ser Gln Leu Glu Leu Glu Lys Ile Lys Lys Glu Lys Pro Ser 50
55 60Leu Ser Tyr Arg Asn Phe Pro Val Ser
Glu Gly Ser Glu Thr Val Leu65 70 75
80Leu Val Asn Leu Glu Asn Glu Thr Gly Glu Leu Thr Gly Glu
Met Asn 85 90 95Trp Ser
Arg Gly Leu Ser Leu Glu Lys Ser Ile Ser Pro Val Ala Asp 100
105 110Ser Leu Ile Arg Phe Ser Tyr Arg Glu
Leu Leu Thr Ala Thr Arg Asn 115 120
125Phe Ser Lys Arg Arg Val Leu Gly Arg Gly Ala Cys Ser Tyr Val Phe
130 135 140Lys Gly Arg Ile Gly Ile Trp
Arg Lys Ala Val Ala Ile Lys Arg Leu145 150
155 160Asp Lys Lys Asp Lys Glu Ser Pro Lys Ser Phe Cys
Arg Glu Leu Met 165 170
175Ile Ala Ser Ser Leu Asn Ser Pro Asn Val Val Pro Leu Leu Gly Phe
180 185 190Cys Ile Asp Pro Asp Gln
Gly Leu Phe Leu Val Tyr Lys Tyr Val Ser 195 200
205Gly Gly Ser Leu Glu Arg Phe Leu His Asp Lys Lys Lys Lys
Lys Ser 210 215 220Arg Lys Thr Pro Leu
Asn Leu Pro Trp Ser Thr Arg Tyr Lys Val Ala225 230
235 240Leu Gly Ile Ala Asp Ala Ile Ala Tyr Leu
His Asn Gly Thr Glu Gln 245 250
255Cys Val Val His Arg Asp Ile Lys Pro Ser Asn Ile Leu Leu Ser Ser
260 265 270Asn Lys Ile Pro Lys
Leu Cys Asp Phe Gly Leu Ala Thr Trp Thr Ala 275
280 285Ala Pro Ser Val Pro Phe Leu Cys Lys Thr Val Lys
Gly Thr Phe Gly 290 295 300Tyr Leu Ala
Pro Glu Tyr Phe Gln His Gly Lys Ile Ser Asp Lys Thr305
310 315 320Asp Val Tyr Ala Phe Gly Val
Val Leu Leu Glu Leu Ile Thr Gly Arg 325
330 335Lys Pro Ile Glu Ala Arg Arg Pro Ser Gly Glu Glu
Asn Leu Val Val 340 345 350Trp
Ala Lys Pro Leu Leu His Arg Gly Ile Glu Ala Thr Glu Glu Leu 355
360 365Leu Asp Pro Arg Leu Lys Cys Thr Arg
Lys Asn Ser Ala Ser Met Glu 370 375
380Arg Met Ile Arg Ala Ala Ala Ala Cys Val Ile Asn Glu Glu Ser Arg385
390 395 400Arg Pro Gly Met
Lys Glu Ile Leu Ser Ile Leu Lys Gly Gly Glu Gly 405
410 415Ile Glu Leu Arg Thr Leu Ser Ser Arg Lys
Lys Ser Asn Leu Pro Gly 420 425
430Ile Met Asp Cys Tyr Pro Gln Leu Gln Arg Thr Lys Ser Glu Met Lys
435 440 445Ser His Leu Thr Leu Ala Met
Leu Gly Val Thr Glu Phe Glu Ala Asp 450 455
460Asp Leu Leu46532354PRTOryza sativa 32Met Ala Gly Ser Asp Glu Val
Asn Arg Asn Glu Cys Lys Thr Val Val1 5 10
15Pro Leu His Thr Trp Val Leu Ile Ser Asn Phe Lys Leu
Ser Tyr Asn 20 25 30Ile Leu
Arg Arg Ala Asp Gly Thr Phe Glu Arg Asp Leu Gly Glu Tyr 35
40 45Leu Asp Arg Arg Val Pro Ala Asn Ala Arg
Pro Leu Glu Gly Val Ser 50 55 60Ser
Phe Asp His Ile Ile Asp Gln Ser Val Gly Leu Glu Val Arg Ile65
70 75 80Tyr Arg Ala Ala Ala Glu
Gly Asp Ala Glu Glu Gly Ala Ala Ala Val 85
90 95Thr Arg Pro Ile Leu Glu Phe Leu Thr Asp Ala Pro
Ala Ala Glu Pro 100 105 110Phe
Pro Val Ile Ile Phe Phe His Gly Gly Ser Phe Val His Ser Ser 115
120 125Ala Ser Ser Thr Ile Tyr Asp Ser Leu
Cys Arg Arg Phe Val Lys Leu 130 135
140Ser Lys Gly Val Val Val Ser Val Asn Tyr Arg Arg Ala Pro Glu His145
150 155 160Arg Tyr Pro Cys
Ala Tyr Asp Asp Gly Trp Thr Ala Leu Lys Trp Val 165
170 175Met Ser Gln Pro Phe Met Arg Ser Gly Gly
Asp Ala Gln Ala Arg Val 180 185
190Phe Leu Ser Gly Asp Ser Ser Gly Gly Asn Ile Ala His His Val Ala
195 200 205Val Arg Ala Ala Asp Glu Gly
Val Lys Val Cys Gly Asn Ile Leu Leu 210 215
220Asn Ala Met Phe Gly Gly Thr Glu Arg Thr Glu Ser Glu Arg Arg
Leu225 230 235 240Asp Gly
Lys Tyr Phe Val Thr Leu Gln Asp Arg Asp Trp Tyr Trp Lys
245 250 255Ala Tyr Leu Pro Glu Asp Ala
Asp Arg Asp His Pro Ala Cys Asn Pro 260 265
270Phe Gly Pro Asn Gly Arg Arg Leu Gly Gly Leu Pro Phe Ala
Lys Ser 275 280 285Leu Ile Ile Val
Ser Gly Leu Asp Leu Thr Cys Asp Arg Gln Leu Ala 290
295 300Tyr Ala Asp Ala Leu Arg Glu Asp Gly His His Val
Lys Val Val Gln305 310 315
320Cys Glu Asn Ala Thr Val Gly Phe Tyr Leu Leu Pro Asn Thr Val His
325 330 335Tyr His Glu Val Met
Glu Glu Ile Ser Asp Phe Leu Asn Ala Asn Leu 340
345 350Tyr Tyr33390PRTZea mays 33Met Gln Ser Ala Ala Ala
Ile Gly Leu Leu Arg Pro Cys Ala Ala Arg1 5
10 15Pro Leu Ala Ala Tyr Thr Ser Pro Arg Arg Gly Ala
Gly Ala Cys Ser 20 25 30Gly
Gly Thr Gln Pro Ile Ile Thr Pro Arg Gly Ile Arg Leu Ser Ala 35
40 45Arg Pro Gly Leu Val Pro Ala Ser Pro
Leu Glu Glu Lys Glu Asn Arg 50 55
60Arg Cys Arg Ala Ser Met His Ala Ala Ala Ser Ala Gly Glu Glu Ala65
70 75 80Gly Gly Gly Leu Ala
Lys Thr Leu Gln Leu Gly Ala Leu Phe Gly Leu 85
90 95Trp Tyr Leu Phe Asn Ile Tyr Phe Asn Ile Tyr
Asn Lys Gln Val Leu 100 105
110Lys Val Leu Pro Tyr Pro Ile Asn Ile Thr Thr Val Gln Phe Ala Val
115 120 125Gly Ser Ala Ile Ala Leu Phe
Met Trp Ile Thr Gly Ile His Lys Arg 130 135
140Pro Lys Ile Ser Gly Ala Gln Leu Phe Ala Ile Leu Pro Leu Ala
Ile145 150 155 160Val His
Thr Met Gly Asn Leu Phe Thr Asn Met Ser Leu Gly Lys Val
165 170 175Ala Val Ser Phe Thr His Thr
Ile Lys Ala Met Glu Pro Phe Phe Ser 180 185
190Val Leu Leu Ser Ala Ile Phe Leu Gly Glu Leu Pro Thr Pro
Trp Val 195 200 205Val Leu Ser Leu
Leu Pro Ile Val Gly Gly Val Ala Leu Ala Ser Leu 210
215 220Thr Glu Ala Ser Phe Asn Trp Ala Gly Phe Trp Ser
Ala Met Ala Ser225 230 235
240Asn Val Thr Phe Gln Ser Arg Asn Val Leu Ser Lys Lys Leu Met Val
245 250 255Lys Lys Glu Glu Ser
Leu Asp Asn Ile Asn Leu Phe Ser Ile Ile Thr 260
265 270Val Met Ser Phe Phe Leu Leu Ala Pro Val Thr Leu
Leu Thr Glu Gly 275 280 285Val Lys
Val Ser Pro Ala Val Leu Gln Ser Ala Gly Leu Asn Leu Lys 290
295 300Gln Val Tyr Thr Arg Ser Leu Ile Ala Ala Phe
Cys Phe His Ala Tyr305 310 315
320Gln Gln Val Ser Tyr Met Ile Leu Ala Arg Val Ser Pro Val Thr His
325 330 335Ser Val Gly Asn
Cys Val Lys Arg Val Val Val Ile Val Thr Ser Val 340
345 350Leu Phe Phe Arg Thr Pro Val Ser Pro Ile Asn
Ser Leu Gly Thr Gly 355 360 365Ile
Ala Leu Ala Gly Val Phe Leu Tyr Ser Gln Leu Lys Arg Leu Lys 370
375 380Pro Lys Pro Lys Thr Ala385
39034477PRTArabidopsis thaliana 34Met Ala His Leu Leu Ser Ala Ser Cys
Pro Ser Val Ile Ser Leu Ser1 5 10
15Ser Ser Ser Ser Lys Asn Ser Val Lys Pro Phe Val Ser Gly Gln
Thr 20 25 30Phe Phe Asn Ala
Gln Leu Leu Ser Arg Ser Ser Leu Lys Gly Leu Leu 35
40 45Phe Gln Glu Lys Lys Pro Arg Lys Ser Cys Val Phe
Arg Ala Thr Ala 50 55 60Val Pro Ile
Thr Gln Gln Ala Pro Pro Glu Thr Ser Thr Asn Asn Ser65 70
75 80Ser Ser Lys Pro Lys Arg Val Met
Val Ile Gly Gly Asp Gly Tyr Cys 85 90
95Gly Trp Ala Thr Ala Leu His Leu Ser Lys Lys Asn Tyr Glu
Val Cys 100 105 110Ile Val Asp
Asn Leu Val Arg Arg Leu Phe Asp His Gln Leu Gly Leu 115
120 125Glu Ser Leu Thr Pro Ile Ala Ser Ile His Asp
Arg Ile Ser Arg Trp 130 135 140Lys Ala
Leu Thr Gly Lys Ser Ile Glu Leu Tyr Val Gly Asp Ile Cys145
150 155 160Asp Phe Glu Phe Leu Ala Glu
Ser Phe Lys Ser Phe Glu Pro Asp Ser 165
170 175Val Val His Phe Gly Glu Gln Arg Ser Ala Pro Tyr
Ser Met Ile Asp 180 185 190Arg
Ser Arg Ala Val Tyr Thr Gln His Asn Asn Val Ile Gly Thr Leu 195
200 205Asn Val Leu Phe Ala Ile Lys Glu Phe
Gly Glu Glu Cys His Leu Val 210 215
220Lys Leu Gly Thr Met Gly Glu Tyr Gly Thr Pro Asn Ile Asp Ile Glu225
230 235 240Glu Gly Tyr Ile
Thr Ile Thr His Asn Gly Arg Thr Asp Thr Leu Pro 245
250 255Tyr Pro Lys Gln Ala Ser Ser Phe Tyr His
Leu Ser Lys Val His Asp 260 265
270Ser His Asn Ile Ala Phe Thr Cys Lys Ala Trp Gly Ile Arg Ala Thr
275 280 285Asp Leu Asn Gln Gly Val Val
Tyr Gly Val Lys Thr Asp Glu Thr Glu 290 295
300Met His Glu Glu Leu Arg Asn Arg Leu Asp Tyr Asp Ala Val Phe
Gly305 310 315 320Thr Ala
Leu Asn Arg Phe Cys Val Gln Ala Ala Val Gly His Pro Leu
325 330 335Thr Val Tyr Gly Lys Gly Gly
Gln Thr Arg Gly Tyr Leu Asp Ile Arg 340 345
350Asp Thr Val Gln Cys Val Glu Ile Ala Ile Ala Asn Pro Ala
Lys Ala 355 360 365Gly Glu Phe Arg
Val Phe Asn Gln Phe Thr Glu Gln Phe Ser Val Asn 370
375 380Glu Leu Ala Ser Leu Val Thr Lys Ala Gly Ser Lys
Leu Gly Leu Asp385 390 395
400Val Lys Lys Met Thr Val Pro Asn Pro Arg Val Glu Ala Glu Glu His
405 410 415Tyr Tyr Asn Ala Lys
His Thr Lys Leu Met Glu Leu Gly Leu Glu Pro 420
425 430His Tyr Leu Ser Asp Ser Leu Leu Asp Ser Leu Leu
Asn Phe Ala Val 435 440 445Gln Phe
Lys Asp Arg Val Asp Thr Lys Gln Ile Met Pro Ser Val Ser 450
455 460Trp Lys Lys Ile Gly Val Lys Thr Lys Ser Met
Thr Thr465 470 47535504PRTArabidopsis
thaliana 35Met Val Ser Leu Leu Ser Phe Phe Leu Leu Leu Leu Val Pro Ile
Phe1 5 10 15Phe Leu Leu
Ile Phe Thr Lys Lys Ile Lys Glu Ser Lys Gln Asn Leu 20
25 30Pro Pro Gly Pro Ala Lys Leu Pro Ile Ile
Gly Asn Leu His Gln Leu 35 40
45Gln Gly Leu Leu His Lys Cys Leu His Asp Leu Ser Lys Lys His Gly 50
55 60Pro Val Met His Leu Arg Leu Gly Phe
Ala Pro Met Val Val Ile Ser65 70 75
80Ser Ser Glu Ala Ala Glu Glu Ala Leu Lys Thr His Asp Leu
Glu Cys 85 90 95Cys Ser
Arg Pro Ile Thr Met Ala Ser Arg Val Phe Ser Arg Asn Gly 100
105 110Lys Asp Ile Gly Phe Gly Val Tyr Gly
Asp Glu Trp Arg Glu Leu Arg 115 120
125Lys Leu Ser Val Arg Glu Phe Phe Ser Val Lys Lys Val Gln Ser Phe
130 135 140Lys Tyr Ile Arg Glu Glu Glu
Asn Asp Leu Met Ile Lys Lys Leu Lys145 150
155 160Glu Leu Ala Ser Lys Gln Ser Pro Val Asp Leu Ser
Lys Ile Leu Phe 165 170
175Gly Leu Thr Ala Ser Ile Ile Phe Arg Thr Ala Phe Gly Gln Ser Phe
180 185 190Phe Asp Asn Lys His Val
Asp Gln Glu Ser Ile Lys Glu Leu Met Phe 195 200
205Glu Ser Leu Ser Asn Met Thr Phe Arg Phe Ser Asp Phe Phe
Pro Thr 210 215 220Ala Gly Leu Lys Trp
Phe Ile Gly Phe Val Ser Gly Gln His Lys Arg225 230
235 240Leu Tyr Asn Val Phe Asn Arg Val Asp Thr
Phe Phe Asn His Ile Val 245 250
255Asp Asp His His Ser Lys Lys Ala Thr Gln Asp Arg Pro Asp Met Val
260 265 270Asp Ala Ile Leu Asp
Met Ile Asp Asn Glu Gln Gln Tyr Ala Ser Phe 275
280 285Lys Leu Thr Val Asp His Leu Lys Gly Val Leu Ser
Asn Ile Tyr His 290 295 300Ala Gly Ile
Asp Thr Ser Ala Ile Thr Leu Ile Trp Ala Met Ala Glu305
310 315 320Leu Val Arg Asn Pro Arg Val
Met Lys Lys Ala Gln Asp Glu Ile Arg 325
330 335Thr Cys Ile Gly Ile Lys Gln Glu Gly Arg Ile Met
Glu Glu Asp Leu 340 345 350Asp
Lys Leu Gln Tyr Leu Lys Leu Val Val Lys Glu Thr Leu Arg Leu 355
360 365His Pro Ala Ala Pro Leu Leu Leu Pro
Arg Glu Thr Met Ala Asp Ile 370 375
380Lys Ile Gln Gly Tyr Asp Ile Pro Gln Lys Arg Ala Leu Leu Val Asn385
390 395 400Ala Trp Ser Ile
Gly Arg Asp Pro Glu Ser Trp Lys Asn Pro Glu Glu 405
410 415Phe Asn Pro Glu Arg Phe Ile Asp Cys Pro
Val Asp Tyr Lys Gly His 420 425
430Ser Phe Glu Leu Leu Pro Phe Gly Ser Gly Arg Arg Ile Cys Pro Gly
435 440 445Ile Ala Met Ala Ile Ala Thr
Ile Glu Leu Gly Leu Leu Asn Leu Leu 450 455
460Tyr Phe Phe Asp Trp Asn Met Pro Glu Lys Lys Lys Asp Met Asp
Met465 470 475 480Glu Glu
Ala Gly Asp Leu Thr Val Asp Lys Lys Val Pro Leu Glu Leu
485 490 495Leu Pro Val Ile Arg Ile Ser
Leu 50036389PRTPseudomonas syringae pv. tomato str. DC3000
36Met Val Met Thr Val Leu Lys Met Thr Asp Leu Asp Leu Gln Gly Lys1
5 10 15Arg Val Leu Ile Arg Glu
Asp Leu Asn Val Pro Ile Lys Asp Gly Val 20 25
30Val Ser Ser Asp Ala Arg Ile Leu Ala Ser Leu Pro Thr
Ile Arg Leu 35 40 45Ala Leu Glu
Lys Gly Ala Ala Val Met Val Cys Ser His Leu Gly Arg 50
55 60Pro Thr Glu Gly Glu Phe Ser Ala Glu Asn Ser Leu
Lys Pro Val Ala65 70 75
80Glu Tyr Leu Ser Lys Ala Leu Gly Arg Asp Val Pro Leu Val Ala Asp
85 90 95Tyr Leu Asp Gly Val Asp
Val Lys Ala Gly Asp Ile Val Leu Phe Glu 100
105 110Asn Val Arg Phe Asn Lys Gly Glu Lys Lys Asn Ala
Asp Glu Leu Ala 115 120 125Gln Lys
Tyr Ala Ala Leu Cys Asp Val Phe Val Met Asp Ala Phe Gly 130
135 140Thr Ala His Arg Ala Glu Gly Ser Thr His Gly
Val Ala Lys Tyr Ala145 150 155
160Lys Val Ala Ala Ala Gly Pro Leu Leu Ala Ala Glu Leu Glu Ala Leu
165 170 175Gly Lys Ala Leu
Gly Ala Pro Ala Gln Pro Met Ala Ala Ile Val Ala 180
185 190Gly Ser Lys Val Ser Thr Lys Leu Asp Val Leu
Asn Ser Leu Ser Ala 195 200 205Ile
Cys Asp Gln Leu Ile Val Gly Gly Gly Ile Ala Asn Thr Phe Leu 210
215 220Ala Ala Ala Gly His Lys Val Gly Lys Ser
Leu Tyr Glu Pro Asp Leu225 230 235
240Leu Asp Thr Ala Arg Ala Ile Ala Ala Lys Val Ser Val Pro Leu
Pro 245 250 255Thr Asp Val
Val Val Ala Lys Glu Phe Ala Glu Ser Ala Thr Ala Thr 260
265 270Val Lys Leu Ile Ala Asp Val Ala Asp Asp
Asp Met Ile Leu Asp Ile 275 280
285Gly Pro Gln Thr Ala Ala His Phe Ala Glu Leu Leu Lys Ser Ser Gly 290
295 300Thr Ile Leu Trp Asn Gly Pro Val
Gly Val Phe Glu Phe Asp Gln Phe305 310
315 320Gly Glu Gly Thr Lys Thr Leu Ala Lys Ala Ile Ala
Glu Ser Lys Ala 325 330
335Phe Ser Ile Ala Gly Gly Gly Asp Thr Leu Ala Ala Ile Asp Lys Tyr
340 345 350Gly Val Ala Asp Gln Ile
Ser Tyr Ile Ser Thr Gly Gly Gly Ala Phe 355 360
365Leu Glu Phe Val Glu Gly Lys Val Leu Pro Ala Val Glu Met
Leu Glu 370 375 380Gln Arg Ala Arg
Ala38537395PRTZea mays 37Met Ser Leu Ile Arg Gly Met Gly Asn Val Ala Lys
Arg Trp Lys Glu1 5 10
15Leu Asn Gly Leu Asn Tyr Trp Lys Gly Leu Val Asp Pro Leu Asp Leu
20 25 30Asp Leu Arg Arg Asn Ile Ile
Asn Tyr Gly Glu Leu Ser Gln Ala Thr 35 40
45Tyr Thr Gly Leu Asn Arg Glu Arg Arg Ser Arg Tyr Ala Gly Ser
Cys 50 55 60Leu Phe Asn Arg Arg Asp
Phe Leu Ser Arg Val Asp Val Ser Asn Pro65 70
75 80Asn Leu Tyr Glu Ile Thr Lys Phe Ile Tyr Ala
Met Cys Thr Val Ser 85 90
95Leu Pro Asp Gly Phe Met Val Lys Ser Leu Ser Lys Ala Ala Trp Ser
100 105 110Arg Gln Ser Asn Trp Met
Gly Phe Val Ala Val Ala Thr Asp Glu Gly 115 120
125Lys Glu Leu Leu Gly Arg Arg Asp Val Val Val Ala Trp Arg
Gly Thr 130 135 140Ile Arg Met Val Glu
Trp Val Asp Asp Leu Asp Ile Ser Leu Val Pro145 150
155 160Ala Ser Glu Ile Val Leu Pro Gly Ser Ala
Ala Asn Pro Cys Val His 165 170
175Gly Gly Trp Leu Ser Val Tyr Thr Ser Ala Asp Pro Gly Ser Gln Tyr
180 185 190Asn Lys Glu Ser Ala
Arg His Gln Val Leu Asn Glu Val Lys Arg Ile 195
200 205Gln Asp Leu Tyr Lys Pro Glu Glu Thr Ser Ile Thr
Ile Thr Gly His 210 215 220Ser Leu Gly
Ala Ala Leu Ala Thr Ile Asn Ala Thr Asp Ile Val Ser225
230 235 240Asn Gly Tyr Asn Arg Ser Cys
Cys Pro Val Ser Ala Phe Val Phe Gly 245
250 255Ser Pro Arg Val Gly Asn Pro Asp Phe Gln Lys Ala
Phe Asp Ser Ala 260 265 270Ala
Asp Leu Arg Leu Leu Arg Val Arg Asn Ser Pro Asp Val Val Pro 275
280 285Lys Trp Pro Lys Leu Gly Tyr Ser Asp
Val Gly Thr Glu Leu Met Ile 290 295
300Asp Thr Gly Glu Ser Pro Tyr Leu Lys Ala Pro Gly Asn Pro Leu Thr305
310 315 320Trp His Asp Met
Glu Cys Tyr Met His Gly Val Ala Gly Ala Gln Gly 325
330 335Ser Ser Gly Gly Phe Glu Leu Leu Val Asp
Arg Asp Val Ala Leu Val 340 345
350Asn Lys His Glu Asp Ala Leu Arg Asn Glu Phe Ala Val Pro Pro Ser
355 360 365Trp Trp Val Val Gln Asn Lys
Gly Met Val Lys Gly Lys Asp Gly Arg 370 375
380Trp His Leu Ala Asp His Glu Glu Asp Asp Asp385
390 39538512PRTArabidopsis thaliana 38Met Ala Thr Leu Leu
Ala Thr Pro Ile Phe Ser Pro Leu Ala Ser Ser1 5
10 15Pro Ala Arg Asn Arg Leu Ser Cys Ser Lys Ile
Arg Phe Gly Ser Lys 20 25
30Asn Gly Lys Ile Leu Asn Ser Asp Gly Ala Gln Lys Leu Asn Leu Ser
35 40 45Lys Phe Arg Lys Pro Asp Gly Gln
Arg Phe Leu Gln Met Gly Ser Ser 50 55
60Lys Glu Met Asn Phe Glu Arg Lys Leu Ser Val Gln Ala Met Asp Gly65
70 75 80Ala Gly Thr Gly Asn
Thr Ser Thr Ile Ser Arg Asn Val Ile Ala Ile 85
90 95Ser His Leu Leu Val Ser Leu Gly Ile Ile Leu
Ala Ala Asp Tyr Phe 100 105
110Leu Lys Gln Ala Phe Val Ala Ala Ser Ile Lys Phe Pro Ser Ala Leu
115 120 125Phe Gly Met Phe Cys Ile Phe
Ser Val Leu Met Ile Phe Asp Ser Val 130 135
140Val Pro Ala Ala Ala Asn Gly Leu Met Asn Phe Phe Glu Pro Ala
Phe145 150 155 160Leu Phe
Ile Gln Arg Trp Leu Pro Leu Phe Tyr Val Pro Ser Leu Val
165 170 175Val Leu Pro Leu Ser Val Arg
Asp Ile Pro Ala Ala Ser Gly Val Lys 180 185
190Ile Cys Tyr Ile Val Ala Gly Gly Trp Leu Ala Ser Leu Cys
Val Ala 195 200 205Gly Tyr Thr Ala
Ile Ala Val Arg Lys Met Val Lys Thr Glu Met Thr 210
215 220Glu Ala Glu Pro Met Ala Lys Pro Ser Pro Phe Ser
Thr Leu Glu Leu225 230 235
240Trp Ser Trp Ser Gly Ile Phe Val Val Ser Phe Val Gly Ala Leu Phe
245 250 255Tyr Pro Asn Ser Leu
Gly Thr Ser Ala Arg Thr Ser Leu Pro Phe Leu 260
265 270Leu Ser Ser Thr Val Leu Gly Tyr Ile Val Gly Ser
Gly Leu Pro Ser 275 280 285Ser Ile
Lys Lys Val Phe His Pro Ile Ile Cys Cys Ala Leu Ser Ala 290
295 300Val Leu Ala Ala Leu Ala Phe Gly Tyr Ala Ser
Gly Ser Gly Leu Asp305 310 315
320Pro Val Leu Gly Asn Tyr Leu Thr Lys Val Ala Ser Asp Pro Gly Ala
325 330 335Gly Asp Ile Leu
Met Gly Phe Leu Gly Ser Val Ile Leu Ser Phe Ala 340
345 350Phe Ser Met Phe Lys Gln Arg Lys Leu Val Lys
Arg His Ala Ala Glu 355 360 365Ile
Phe Thr Ser Val Ile Val Ser Thr Val Phe Ser Leu Tyr Ser Thr 370
375 380Ala Leu Val Gly Arg Leu Val Gly Leu Glu
Pro Ser Leu Thr Val Ser385 390 395
400Ile Leu Pro Arg Cys Ile Thr Val Ala Leu Ala Leu Ser Ile Val
Ser 405 410 415Leu Phe Glu
Gly Thr Asn Ser Ser Leu Thr Ala Ala Val Val Val Val 420
425 430Thr Gly Leu Ile Gly Ala Asn Phe Val Gln
Val Val Leu Asp Lys Leu 435 440
445Arg Leu Arg Asp Pro Ile Ala Arg Gly Ile Ala Thr Ala Ser Ser Ala 450
455 460His Gly Leu Gly Thr Ala Ala Leu
Ser Ala Lys Glu Pro Glu Ala Leu465 470
475 480Pro Phe Cys Ala Ile Ala Tyr Ala Leu Thr Gly Ile
Phe Gly Ser Leu 485 490
495Leu Cys Ser Val Pro Ala Val Arg Gln Ser Leu Leu Ala Val Val Gly
500 505 51039311PRTZea mays 39Met Ala
Arg Asn Glu Glu Lys Ala Gln Ser Met Leu Asn Arg Phe Ile1 5
10 15Thr Met Lys Gln Glu Glu Lys Arg
Lys Pro Arg Glu Arg Arg Pro Tyr 20 25
30Leu Ala Ser Glu Cys Arg Asp Leu Ala Asp Ala Glu Arg Trp Arg
Ser 35 40 45Glu Ile Leu Arg Glu
Ile Gly Ala Lys Val Ala Glu Ile Gln Asn Glu 50 55
60Gly Leu Gly Glu His Arg Leu Arg Asp Leu Asn Asp Glu Ile
Asn Lys65 70 75 80Leu
Leu Arg Glu Arg Gly His Trp Glu Arg Arg Ile Val Glu Leu Gly
85 90 95Gly Arg Asp Tyr Ser Arg Ser
Ser Asn Ala Pro Leu Met Thr Asp Leu 100 105
110Asp Gly Asn Ile Val Ala Val Pro Asn Pro Ser Gly Arg Gly
Pro Gly 115 120 125Tyr Arg Tyr Phe
Gly Ala Ala Arg Lys Leu Pro Gly Val Arg Glu Leu 130
135 140Phe Asp Lys Pro Pro Glu Met Arg Lys Arg Arg Thr
Arg Tyr Glu Ile145 150 155
160His Lys Arg Ile Asn Ala Gly Tyr Tyr Gly Tyr Tyr Asp Asp Glu Asp
165 170 175Gly Val Leu Glu Arg
Leu Glu Gly Pro Ala Gly Lys Arg Met Arg Glu 180
185 190Glu Ile Val Ser Glu Trp His Arg Val Glu Arg Val
Arg Arg Glu Ala 195 200 205Met Lys
Gly Val Met Ser Gly Glu Val Ala Ala Ala Gly Gly Arg Ser 210
215 220Gly Glu Ala Ala Arg Glu Val Leu Phe Glu Gly
Val Glu Glu Glu Val225 230 235
240Glu Glu Glu Arg Lys Arg Glu Glu Glu Lys Arg Glu Arg Glu Lys Gly
245 250 255Glu Glu Val Gly
Arg Glu Phe Val Ala His Val Pro Leu Pro Asp Glu 260
265 270Lys Glu Ile Glu Arg Met Val Leu Glu Arg Lys
Lys Lys Glu Leu Leu 275 280 285Ser
Lys Tyr Ala Ser Asp Ser Leu Leu Val Glu Gln Glu Glu Ala Lys 290
295 300Glu Met Leu Asn Val Arg Arg305
31040682PRTZea mays 40Met Asp Leu Ala Arg Arg Gly Gly Ala Ala Gly
Ala Asp Asp Glu Gly1 5 10
15Glu Ile Glu Arg His Glu Pro Ala Pro Glu Asp Met Glu Ser Asp Pro
20 25 30Ala Ala Ala Arg Glu Lys Glu
Leu Glu Leu Glu Arg Val Gln Ser Trp 35 40
45Arg Glu Gln Val Thr Leu Arg Gly Val Val Ala Ala Leu Leu Ile
Gly 50 55 60Phe Met Tyr Ser Val Ile
Val Met Lys Ile Ala Leu Thr Thr Gly Leu65 70
75 80Val Pro Thr Leu Asn Val Ser Ala Ala Leu Met
Ala Phe Leu Ala Leu 85 90
95Arg Gly Trp Thr Arg Val Leu Glu Arg Leu Gly Val Ala His Arg Pro
100 105 110Phe Thr Arg Gln Glu Asn
Cys Val Ile Glu Thr Cys Ala Val Ala Cys 115 120
125Tyr Thr Ile Ala Phe Gly Gly Gly Phe Gly Ser Thr Leu Leu
Gly Leu 130 135 140Asp Lys Lys Thr Tyr
Glu Leu Ala Gly Ala Ser Pro Ala Asn Val Pro145 150
155 160Gly Ser Tyr Lys Asp Pro Gly Phe Gly Trp
Met Ala Gly Phe Val Ala 165 170
175Ala Ile Ser Phe Ala Gly Leu Leu Ser Leu Ile Pro Leu Arg Lys Val
180 185 190Leu Val Ile Asp Tyr
Lys Leu Thr Tyr Pro Ser Gly Thr Ala Thr Ala 195
200 205Val Leu Ile Asn Gly Phe His Thr Lys Gln Gly Asp
Lys Asn Ala Arg 210 215 220Met Gln Val
Arg Gly Phe Leu Lys Tyr Phe Gly Leu Ser Phe Val Trp225
230 235 240Ser Phe Phe Gln Trp Phe Tyr
Thr Gly Gly Glu Val Cys Gly Phe Val 245
250 255Gln Phe Pro Thr Phe Gly Leu Lys Ala Trp Lys Gln
Thr Phe Phe Phe 260 265 270Asp
Phe Ser Leu Thr Tyr Val Gly Ala Gly Met Ile Cys Ser His Leu 275
280 285Val Asn Ile Ser Thr Leu Leu Gly Ala
Ile Leu Ser Trp Gly Ile Leu 290 295
300Trp Pro Leu Ile Ser Lys Gln Lys Gly Glu Trp Tyr Pro Ala Asn Ile305
310 315 320Pro Glu Ser Ser
Met Lys Ser Leu Tyr Gly Tyr Lys Ala Phe Leu Cys 325
330 335Ile Ala Leu Ile Met Gly Asp Gly Thr Tyr
His Phe Phe Lys Val Phe 340 345
350Gly Val Thr Val Lys Ser Leu His Gln Arg Leu Ser Arg Lys Arg Ala
355 360 365Thr Asn Arg Val Ala Asn Gly
Gly Asp Glu Met Ala Ala Leu Asp Asp 370 375
380Leu Gln Arg Asp Glu Ile Phe Ser Asp Gly Ser Phe Pro Ala Trp
Ala385 390 395 400Ala Tyr
Ala Gly Tyr Ala Ala Leu Thr Val Val Ser Ala Val Ile Ile
405 410 415Pro His Met Phe Arg Gln Val
Lys Trp Tyr Tyr Val Ile Val Ala Tyr 420 425
430Val Leu Ala Pro Leu Leu Gly Phe Ala Asn Ser Tyr Gly Thr
Gly Leu 435 440 445Thr Asp Ile Asn
Met Ala Tyr Asn Tyr Gly Lys Ile Ala Leu Phe Ile 450
455 460Phe Ala Ala Trp Ala Gly Arg Asp Asn Gly Val Ile
Ala Gly Leu Ala465 470 475
480Gly Gly Thr Leu Val Lys Gln Leu Val Met Ala Ser Ala Asp Leu Met
485 490 495His Asp Phe Lys Thr
Gly His Leu Thr Met Thr Ser Pro Arg Ser Leu 500
505 510Leu Val Ala Gln Phe Ile Gly Thr Ala Met Gly Cys
Val Val Ala Pro 515 520 525Leu Thr
Phe Leu Leu Phe Tyr Asn Ala Phe Asp Ile Gly Asn Pro Thr 530
535 540Gly Tyr Trp Lys Ala Pro Tyr Gly Leu Ile Tyr
Arg Asn Met Ala Ile545 550 555
560Leu Gly Val Glu Gly Phe Ser Val Leu Pro Arg His Cys Leu Ala Leu
565 570 575Ser Ala Gly Phe
Phe Ala Phe Ala Phe Val Phe Ser Val Ala Arg Asp 580
585 590Val Leu Pro Arg Lys Tyr Ala Arg Phe Val Pro
Leu Pro Met Ala Met 595 600 605Ala
Val Pro Phe Leu Val Gly Gly Ser Phe Ala Ile Asp Met Cys Val 610
615 620Gly Ser Leu Ala Val Phe Val Trp Glu Lys
Val Asn Arg Lys Glu Ala625 630 635
640Val Phe Met Val Pro Ala Val Ala Ser Gly Leu Ile Cys Gly Asp
Gly 645 650 655Ile Trp Thr
Phe Pro Ser Ser Ile Leu Ala Leu Ala Lys Ile Lys Pro 660
665 670Pro Ile Cys Met Lys Phe Thr Pro Gly Ser
675 68041453PRTArabidopsis thaliana 41Met Ala Lys
Val Tyr Trp Pro Tyr Phe Asp Pro Glu Tyr Glu Asn Leu1 5
10 15Ser Ser Arg Ile Asn Pro Pro Ser Val
Ser Ile Asp Asn Thr Ser Cys 20 25
30Lys Glu Cys Thr Leu Val Lys Val Asp Ser Met Asn Lys Pro Gly Ile
35 40 45Leu Leu Glu Val Val Gln Val
Leu Thr Asp Leu Asp Leu Thr Ile Thr 50 55
60Lys Ala Tyr Ile Ser Ser Asp Gly Gly Trp Phe Met Asp Val Phe His65
70 75 80Val Thr Asp Gln
Gln Gly Asn Lys Val Thr Asp Ser Lys Thr Ile Asp 85
90 95Tyr Ile Glu Lys Val Leu Gly Pro Lys Gly
His Ala Ser Ala Ser Gln 100 105
110Asn Thr Trp Pro Gly Lys Arg Val Gly Val His Ser Leu Gly Asp His
115 120 125Thr Ser Ile Glu Ile Ile Ala
Arg Asp Arg Pro Gly Leu Leu Ser Glu 130 135
140Val Ser Ala Val Leu Ala Asp Leu Asn Ile Asn Val Val Ala Ala
Glu145 150 155 160Ala Trp
Thr His Asn Arg Arg Ile Ala Cys Val Leu Tyr Val Asn Asp
165 170 175Asn Ala Thr Ser Arg Ala Val
Asp Asp Pro Glu Arg Leu Ser Ser Met 180 185
190Glu Glu Gln Leu Asn Asn Val Leu Arg Gly Cys Glu Glu Gln
Asp Glu 195 200 205Lys Phe Ala Arg
Thr Ser Leu Ser Ile Gly Ser Thr His Val Asp Arg 210
215 220Arg Leu His Gln Met Phe Phe Ala Asp Arg Asp Tyr
Glu Ala Val Thr225 230 235
240Lys Leu Asp Asp Ser Ala Ser Cys Gly Phe Glu Pro Lys Ile Thr Val
245 250 255Glu His Cys Glu Glu
Lys Gly Tyr Ser Val Ile Asn Val Ser Cys Glu 260
265 270Asp Arg Pro Lys Leu Met Phe Asp Ile Val Cys Thr
Leu Thr Asp Met 275 280 285Gln Tyr
Ile Val Phe His Ala Thr Ile Ser Ser Ser Gly Ser His Ala 290
295 300Ser Gln Glu Tyr Phe Ile Arg His Lys Asp Gly
Cys Thr Leu Asp Thr305 310 315
320Glu Gly Glu Lys Glu Arg Val Val Lys Cys Leu Glu Ala Ala Ile His
325 330 335Arg Arg Val Ser
Glu Gly Trp Ser Leu Glu Leu Cys Ala Lys Asp Arg 340
345 350Val Gly Leu Leu Ser Glu Val Thr Arg Ile Leu
Arg Glu His Gly Leu 355 360 365Ser
Val Ser Arg Ala Gly Val Thr Thr Val Gly Glu Gln Ala Val Asn 370
375 380Val Phe Tyr Val Lys Asp Ala Ser Gly Asn
Pro Val Asp Val Lys Thr385 390 395
400Ile Glu Ala Leu Arg Gly Glu Ile Gly His Ser Met Met Ile Asp
Phe 405 410 415Lys Asn Lys
Val Pro Ser Arg Lys Trp Lys Glu Glu Gly Gln Ala Gly 420
425 430Thr Gly Gly Gly Trp Ala Lys Thr Ser Phe
Phe Phe Gly Asn Leu Leu 435 440
445Glu Lys Leu Leu Pro 45042334PRTEscherichia coli 42Met Ala Gln Val
Ser Arg Ile Cys Asn Gly Val Gln Asn Pro Ser Leu1 5
10 15Ile Ser Asn Leu Ser Lys Ser Ser Gln Arg
Lys Ser Pro Leu Ser Val 20 25
30Ser Leu Lys Thr Gln Gln His Pro Arg Ala Tyr Pro Ile Ser Ser Ser
35 40 45Trp Gly Leu Lys Lys Ser Gly Met
Thr Leu Ile Gly Ser Glu Leu Arg 50 55
60Pro Leu Lys Val Met Ser Ser Val Ser Thr Ala Cys Met Met Asn Pro65
70 75 80Leu Ile Ile Lys Leu
Gly Gly Val Leu Leu Asp Ser Glu Glu Ala Leu 85
90 95Glu Arg Leu Phe Ser Ala Leu Val Asn Tyr Arg
Glu Ser His Gln Arg 100 105
110Pro Leu Val Ile Val His Gly Gly Gly Cys Val Val Asp Glu Leu Met
115 120 125Lys Gly Leu Asn Leu Pro Val
Lys Lys Lys Asn Gly Leu Arg Val Thr 130 135
140Pro Ala Asp Gln Ile Asp Ile Ile Thr Gly Ala Leu Ala Gly Thr
Ala145 150 155 160Asn Lys
Thr Leu Leu Ala Trp Ala Lys Lys His Gln Ile Ala Ala Val
165 170 175Gly Leu Phe Leu Gly Asp Gly
Asp Ser Val Lys Val Thr Gln Leu Asp 180 185
190Glu Glu Leu Gly His Val Gly Leu Ala Gln Pro Gly Ser Pro
Lys Leu 195 200 205Ile Asn Ser Leu
Leu Glu Asn Gly Tyr Leu Pro Val Val Ser Ser Ile 210
215 220Gly Val Thr Asp Glu Gly Gln Leu Met Asn Val Asn
Ala Asp Gln Ala225 230 235
240Ala Thr Ala Leu Ala Ala Thr Leu Gly Ala Asp Leu Ile Leu Leu Ser
245 250 255Asp Val Ser Gly Ile
Leu Asp Gly Lys Gly Gln Arg Ile Ala Glu Met 260
265 270Thr Ala Ala Lys Ala Glu Gln Leu Ile Glu Gln Gly
Ile Ile Thr Asp 275 280 285Gly Met
Ile Val Lys Val Asn Ala Ala Leu Asp Ala Ala Arg Thr Leu 290
295 300Gly Arg Pro Val Asp Ile Ala Ser Trp Arg His
Ala Glu Gln Leu Pro305 310 315
320Ala Leu Phe Asn Gly Met Pro Met Gly Thr Arg Ile Leu Ala
325 33043557PRTGlycine max 43Met Ala Leu Lys Thr Leu
Ser Thr Phe Leu Ser Pro Leu Ser Leu Pro1 5
10 15Asn Thr Lys Phe Pro Gln Phe Leu Thr Thr Lys Pro
Ser Leu Ile Leu 20 25 30Cys
Glu Phe Pro Arg Ser Gln Lys Ser Arg Leu Leu Ala Ala Asp Ser 35
40 45Glu Gly Thr Gly Ala Ala Ala Pro Ser
Pro Gly Glu Lys Phe Leu Glu 50 55
60Arg Gln Gln Ser Phe Glu Asp Ala Lys Ile Ile Leu Lys Glu Asn Lys65
70 75 80Lys Lys Arg Lys Lys
Lys Asp Asn Ala Ile Lys Ala Ser Arg Ala Val 85
90 95Ala Ser Cys Tyr Gly Cys Gly Ala Pro Leu His
Thr Ser Asp Ala Asp 100 105
110Ala Pro Gly Tyr Val Asp Pro Glu Thr Tyr Glu Leu Lys Lys Lys His
115 120 125His Gln Leu Arg Thr Val Leu
Cys Arg Arg Cys Arg Leu Leu Ser His 130 135
140Gly Lys Met Ile Thr Ala Val Gly Gly His Gly Gly Tyr Pro Gly
Gly145 150 155 160Lys Leu
Phe Val Thr Ala Glu Glu Leu Arg Glu Lys Leu Ser His Leu
165 170 175Arg His Glu Lys Ala Leu Ile
Val Lys Leu Val Asp Ile Val Asp Phe 180 185
190Asn Gly Ser Phe Leu Ser Arg Val Arg Asp Leu Ala Gly Ser
Asn Pro 195 200 205Ile Ile Leu Val
Val Thr Lys Val Asp Leu Leu Pro Arg Asp Thr Asp 210
215 220Leu His Cys Val Gly Asp Trp Val Val Glu Ala Thr
Met Arg Lys Lys225 230 235
240Leu Asn Val Leu Ser Val His Leu Thr Ser Ser Lys Ser Leu Val Gly
245 250 255Ile Thr Gly Val Ile
Ser Glu Ile Gln Lys Glu Lys Lys Gly Arg Asp 260
265 270Val Tyr Ile Leu Gly Ser Ala Asn Val Gly Lys Ser
Ala Phe Ile Asn 275 280 285Ala Leu
Leu Lys Thr Met Ala Ile Asn Asp Pro Val Ala Ala Ser Ala 290
295 300Gln Arg Tyr Lys Pro Ile Gln Ser Ala Val Pro
Gly Thr Thr Leu Gly305 310 315
320Pro Ile Gln Ile Asn Ala Phe Leu Gly Gly Gly Lys Leu Tyr Asp Thr
325 330 335Pro Gly Val His
Leu Tyr His Arg Gln Thr Ala Val Val His Ser Glu 340
345 350Asp Leu Pro Ile Leu Ala Pro Gln Ser Arg Leu
Arg Gly Leu Ser Phe 355 360 365Pro
Ser Ser Ile Leu Ser Ser Val Glu Glu Gly Ala Ser Thr Ile Val 370
375 380Asn Gly Leu Asn Ala Phe Ser Ile Phe Trp
Gly Gly Leu Val Arg Ile385 390 395
400Asp Val Leu Lys Val Leu Pro Glu Thr Cys Leu Thr Phe Tyr Gly
Pro 405 410 415Lys Arg Ile
Pro Ile His Met Val Pro Thr Glu Gln Ala Val Glu Phe 420
425 430Tyr Gln Thr Glu Leu Gly Val Leu Leu Thr
Pro Pro Ser Gly Gly Glu 435 440
445Asn Ala Glu Asn Trp Lys Gly Leu Glu Ser Glu Arg Lys Leu Gln Ile 450
455 460Lys Phe Glu Asp Val Asp Ser Tyr
Asp Pro Lys Pro Ala Cys Asp Ile465 470
475 480Ala Ile Ser Gly Leu Gly Trp Phe Thr Val Glu Pro
Val Ser Arg Ser 485 490
495Leu Lys Ile Ser Gln Pro Lys Pro Val Glu Thr Ala Gly Glu Leu Ile
500 505 510Leu Ala Val His Val Pro
Lys Ala Val Glu Ile Phe Val Arg Ser Pro 515 520
525Ile Pro Val Gly Lys Ala Gly Ala Glu Trp Tyr Gln Tyr Val
Glu Leu 530 535 540Thr Glu Lys Gln Glu
Glu Met Arg Pro Lys Trp Tyr Phe545 550
55544426PRTZea mays 44Met Ala Ala Ala Leu Ala Ser Ser Arg Tyr Cys Trp Ser
Arg Pro Ser1 5 10 15Leu
Pro Pro Gln Pro Thr Arg Gly Arg Arg Ser Val Thr Ser Cys Ala 20
25 30Leu Ser Gly Arg Glu Lys Arg Asn
Ser Phe Ser Trp Arg Glu Cys Ala 35 40
45Ile Ser Val Ala Leu Ser Val Gly Leu Ile Thr Gly Ala Pro Thr Phe
50 55 60Gly Pro Pro Ala Tyr Ala Ser Ser
Leu Glu Pro Val Leu Pro Asp Val65 70 75
80Ser Val Leu Ile Ser Gly Pro Pro Ile Lys Asp Pro Gly
Ala Leu Leu 85 90 95Arg
Tyr Ala Leu Pro Ile Asp Asn Lys Ala Ile Arg Glu Val Gln Lys
100 105 110Pro Leu Glu Asp Ile Thr Asp
Ser Leu Lys Val Ala Gly Val Arg Ala 115 120
125Leu Asp Ser Val Glu Arg Asn Val Arg Gln Ala Ser Lys Ala Leu
Asn 130 135 140Asn Gly Arg Ser Leu Ile
Leu Ala Gly Leu Ala Glu Pro Lys Arg Ala145 150
155 160Asn Gly Glu Glu Leu Leu Asn Lys Leu Ala Val
Gly Phe Glu Glu Leu 165 170
175Gln Arg Ile Val Glu Asp Arg Asn Arg Asp Ala Val Ala Pro Lys Gln
180 185 190Lys Glu Leu Leu Gln Tyr
Val Gly Thr Val Glu Glu Asp Met Val Asp 195 200
205Gly Phe Pro Phe Glu Ile Pro Glu Glu Tyr Ser Asn Met Pro
Leu Leu 210 215 220Lys Gly Arg Ala Thr
Val Asp Met Lys Val Lys Ile Lys Asp Asn Pro225 230
235 240Asn Met Glu Asp Cys Val Phe Arg Ile Val
Leu Asp Gly Tyr Asn Ala 245 250
255Pro Val Thr Ala Gly Asn Phe Val Asp Leu Val Lys Arg Lys Phe Tyr
260 265 270Asp Gly Met Glu Ile
Gln Arg Ala Asp Gly Phe Val Val Gln Thr Gly 275
280 285Asp Pro Glu Gly Pro Ala Glu Gly Phe Ile Asp Pro
Ser Thr Gly Lys 290 295 300Ile Arg Thr
Val Pro Leu Glu Ile Met Val Asp Gly Asp Lys Ala Pro305
310 315 320Val Tyr Gly Glu Thr Leu Glu
Glu Leu Gly Arg Tyr Lys Ala Gln Thr 325
330 335Lys Leu Pro Phe Asn Ala Phe Gly Thr Met Ala Met
Ala Arg Glu Glu 340 345 350Phe
Asp Asp Asn Ser Ala Ser Ser Gln Val Phe Trp Leu Leu Lys Glu 355
360 365Ser Glu Leu Thr Pro Ser Asn Ala Asn
Ile Leu Asp Gly Arg Tyr Ala 370 375
380Val Phe Gly Tyr Val Thr Glu Asn Glu Asp Tyr Leu Ala Asp Val Lys385
390 395 400Val Gly Asp Val
Ile Glu Ser Ile Gln Val Val Ser Gly Leu Asp Asn 405
410 415Leu Val Asn Pro Ser Tyr Lys Ile Val Gly
420 42545144PRTArabidopsis thaliana 45Met Met
Gln Glu Leu Gly Leu Gln Arg Phe Ser Asn Asp Val Val Arg1 5
10 15Leu Asp Leu Thr Pro Pro Ser Gln
Thr Ser Ser Thr Ser Leu Ser Ile 20 25
30Asp Glu Glu Glu Ser Thr Glu Ala Lys Ile Arg Arg Leu Ile Ser
Glu 35 40 45His Pro Val Ile Ile
Phe Ser Arg Ser Ser Cys Cys Met Cys His Val 50 55
60Met Lys Arg Leu Leu Ala Thr Ile Gly Val Ile Pro Thr Val
Ile Glu65 70 75 80Leu
Asp Asp His Glu Val Ser Ser Leu Pro Thr Ala Leu Gln Asp Glu
85 90 95Tyr Ser Gly Gly Val Ser Val
Val Gly Pro Pro Pro Ala Val Phe Ile 100 105
110Gly Arg Glu Cys Val Gly Gly Leu Glu Ser Leu Val Ala Leu
His Leu 115 120 125Ser Gly Gln Leu
Val Pro Lys Leu Val Gln Val Gly Ala Leu Trp Val 130
135 14046509PRTEscherichia coli 46Met Ala Ser Ser Met Leu
Ser Ser Ala Thr Met Val Ala Ser Pro Ala1 5
10 15Gln Ala Thr Met Val Ala Pro Phe Asn Gly Leu Lys
Ser Ser Ala Ala 20 25 30Phe
Pro Ala Thr Arg Lys Ala Asn Asn Asp Ile Thr Ser Ile Thr Ser 35
40 45Asn Gly Gly Arg Val Asn Cys Met Gln
Val Trp Pro Pro Ile Gly Lys 50 55
60Lys Lys Phe Glu Thr Leu Ser Tyr Leu Pro Asp Leu Thr Asp Ser Gly65
70 75 80Gly Arg Val Asn Cys
Met Gln Ala Met Ser Asn Asn Glu Phe His Gln 85
90 95Arg Arg Leu Ser Ala Thr Pro Arg Gly Val Gly
Val Met Cys Asn Phe 100 105
110Phe Ala Gln Ser Ala Glu Asn Ala Thr Leu Lys Asp Val Glu Gly Asn
115 120 125Glu Tyr Ile Asp Phe Ala Ala
Gly Ile Ala Val Leu Asn Thr Gly His 130 135
140Arg His Pro Asp Leu Val Ala Ala Val Glu Gln Gln Leu Gln Gln
Phe145 150 155 160Thr His
Thr Ala Tyr Gln Ile Val Pro Tyr Glu Ser Tyr Val Thr Leu
165 170 175Ala Glu Lys Ile Asn Ala Leu
Ala Pro Val Ser Gly Gln Ala Lys Thr 180 185
190Ala Phe Phe Thr Thr Gly Ala Glu Ala Val Glu Asn Ala Val
Lys Ile 195 200 205Ala Arg Ala His
Thr Gly Arg Pro Gly Val Ile Ala Phe Ser Gly Gly 210
215 220Phe His Gly Arg Thr Tyr Met Thr Met Ala Leu Thr
Gly Lys Val Ala225 230 235
240Pro Tyr Lys Ile Gly Phe Gly Pro Phe Pro Gly Ser Val Tyr His Val
245 250 255Pro Tyr Pro Ser Asp
Leu His Gly Ile Ser Thr Gln Asp Ser Leu Asp 260
265 270Ala Ile Glu Arg Leu Phe Lys Ser Asp Ile Glu Ala
Lys Gln Val Ala 275 280 285Ala Ile
Ile Phe Glu Pro Val Gln Gly Glu Gly Gly Phe Asn Val Ala 290
295 300Pro Lys Glu Leu Val Ala Ala Ile Arg Arg Leu
Cys Asp Glu His Gly305 310 315
320Ile Val Met Ile Ala Asp Glu Val Gln Ser Gly Phe Ala Arg Thr Gly
325 330 335Lys Leu Phe Ala
Met Asp His Tyr Ala Asp Lys Pro Asp Leu Met Thr 340
345 350Met Ala Lys Ser Leu Ala Gly Gly Met Pro Leu
Ser Gly Val Val Gly 355 360 365Asn
Ala Asn Ile Met Asp Ala Pro Ala Pro Gly Gly Leu Gly Gly Thr 370
375 380Tyr Ala Gly Asn Pro Leu Ala Val Ala Ala
Ala His Ala Val Leu Asn385 390 395
400Ile Ile Asp Lys Glu Ser Leu Cys Glu Arg Ala Asn Gln Leu Gly
Gln 405 410 415Arg Leu Lys
Asn Thr Leu Ile Asp Ala Lys Glu Ser Val Pro Ala Ile 420
425 430Ala Ala Val Arg Gly Leu Gly Ser Met Ile
Ala Val Glu Phe Asn Asp 435 440
445Pro Gln Thr Gly Glu Pro Ser Ala Ala Ile Ala Gln Lys Ile Gln Gln 450
455 460Arg Ala Leu Ala Gln Gly Leu Leu
Leu Leu Thr Cys Gly Ala Tyr Gly465 470
475 480Asn Val Ile Arg Phe Leu Tyr Pro Leu Thr Ile Pro
Asp Ala Gln Phe 485 490
495Asp Ala Ala Met Lys Ile Leu Gln Asp Ala Leu Ser Asp 500
50547490PRTSynechocystis sp. 47Met Ala Ser Ser Met Leu Ser
Ser Ala Thr Met Val Ala Ser Pro Ala1 5 10
15Gln Ala Thr Met Val Ala Pro Phe Asn Gly Leu Lys Ser
Ser Ala Ala 20 25 30Phe Pro
Ala Thr Arg Lys Ala Asn Asn Asp Ile Thr Ser Ile Thr Ser 35
40 45Asn Gly Gly Arg Val Asn Cys Met Gln Val
Trp Pro Pro Ile Gly Lys 50 55 60Lys
Lys Phe Glu Thr Leu Ser Tyr Leu Pro Asp Leu Thr Asp Ser Gly65
70 75 80Gly Arg Val Asn Cys Met
Gln Ala Met Thr Pro Glu Leu Asn Pro Asn 85
90 95Phe Pro Glu Glu Thr Thr Ser Asp Ala Trp Leu Thr
Pro Ala Asp Ala 100 105 110Gly
Gln Asp Gly Asp Ala Gln Glu Pro Ala Glu Asp Gly Gly Glu Glu 115
120 125Gly Val Val Ser Glu Glu Leu Ala Leu
Pro Glu Asp Leu Pro Pro Met 130 135
140Asp Ala Met Val Ala Ala Val Glu Glu Met Thr Pro Val Val Val Pro145
150 155 160Glu Thr Val Pro
Glu Thr Glu Thr Pro Ala Leu Glu Asp Leu Val Ala 165
170 175Gln Lys Thr Ala Leu Glu Lys Asp Ile Ala
Ala Leu Gln Arg Glu Lys 180 185
190Ala Gln Trp Tyr Gly Gln Gln Phe Gln Gln Leu Gln Arg Glu Met Ala
195 200 205Arg Leu Val Glu Glu Gly Thr
Arg Glu Leu Gly Gln Arg Lys Ala Ala 210 215
220Leu Glu Lys Glu Ile Glu Lys Leu Glu Arg Arg Gln Glu Arg Ile
Gln225 230 235 240Gln Glu
Met Arg Thr Thr Phe Ala Gly Ala Ser Gln Glu Leu Ala Ile
245 250 255Arg Val Gln Gly Phe Lys Asp
Tyr Leu Val Gly Ser Leu Gln Asp Leu 260 265
270Val Ser Ala Ala Asp Gln Leu Glu Leu Gly Val Gly Asp Ser
Trp Glu 275 280 285Ser Ser Ser Thr
His Gly Asp Ala Ile Ile Glu Asn Ala Asp Pro Thr 290
295 300Pro Val Val Ser Phe Ala Glu Gln Gly Phe Ser Ser
Gln Lys Arg Gln305 310 315
320Ile Gln Ala Leu Leu Glu Gln Tyr Arg Thr Arg Pro Asp Tyr Tyr Gly
325 330 335Pro Pro Trp Gln Leu
Arg Arg Thr Phe Glu Pro Val His Ala Glu Arg 340
345 350Ile Glu Asn Trp Phe Phe Thr Leu Gly Gly Arg Gly
Ala Ile Leu Ser 355 360 365Leu Asp
Ser Arg Leu Gln Asn Ile Leu Val Gly Ser Ala Ala Ile Ala 370
375 380Ile Leu Asn Gln Leu Tyr Gly Asp Arg Cys Arg
Ala Leu Ile Leu Ala385 390 395
400Ala Thr Pro Glu Arg Leu Gly Glu Trp Arg Arg Gly Leu Gln Asp Cys
405 410 415Leu Gly Ile Ser
Arg Ser Asp Phe Gly Pro Asp Arg Gly Ile Val Leu 420
425 430Phe Glu Ser Ala Asn Ala Leu Ile Gln Arg Ala
Glu Arg Leu Val Gly 435 440 445Asp
Arg Gln Met Pro Leu Val Leu Val Asp Glu Thr Glu Glu Gln Ile 450
455 460Asp Leu Ala Leu Leu Gln Phe Pro Leu Leu
Leu Ala Phe Ala Pro Ser465 470 475
480Tyr Gln Val Gly Gly Ser Asn Tyr Phe Ser 485
49048404PRTZea mays 48Met Ala Gly Glu Leu Arg His Arg Arg
Ala Pro Ser Glu Asp Glu Gly1 5 10
15Val Ala Ser Ser Gln Arg Leu Asp Ser Ala Pro Ala Gly Asn Gly
Lys 20 25 30Ala Gly Thr Ser
Ser Gly Gly Gly Glu Gly Ala Glu Pro Arg Gly Gly 35
40 45Lys Arg Asp Ala Leu Gly Trp Leu Glu Trp Cys Arg
Gly Trp Met Ala 50 55 60Ile Val Gly
Glu Phe Leu Phe Gln Arg Ile Ala Ala Ser His Leu Ala65 70
75 80Asn Pro Leu Glu Leu Pro Pro Leu
Asp Gly Val Ser Ile Val Val Thr 85 90
95Gly Ala Thr Ser Gly Ile Gly Leu Glu Ile Ala Arg Gln Leu
Ala Leu 100 105 110Ala Gly Ala
His Val Val Met Ala Val Arg Arg Pro Lys Val Ala Gln 115
120 125Glu Leu Ile Gln Lys Trp Gln Asn Glu Asn Ser
Glu Thr Gly Arg Pro 130 135 140Leu Asn
Ala Glu Val Met Glu Leu Asp Leu Leu Ser Leu Asp Ser Val145
150 155 160Val Lys Phe Ala Asp Ala Trp
Asn Ala Arg Met Ala Pro Leu His Val 165
170 175Leu Ile Asn Asn Ala Gly Ile Phe Ala Ile Gly Glu
Pro Gln His Phe 180 185 190Ser
Lys Asp Gly His Glu Glu His Met Gln Val Asn His Leu Ala Pro 195
200 205Ala Leu Leu Ala Met Leu Leu Ile Pro
Ser Leu Leu Arg Gly Ser Pro 210 215
220Ser Arg Ile Val Asn Val Asn Ser Ile Met His Ser Val Gly Phe Val225
230 235 240Asp Ala Glu Asp
Phe Asn Leu Arg Lys His Lys Tyr Arg Ser Trp Leu 245
250 255Ala Tyr Ser Asn Ser Lys Leu Ala Gln Val
Lys Phe Ser Ser Met Leu 260 265
270His Lys Arg Ile Pro Ala Glu Ala Gly Ile Ser Ile Ile Cys Ala Ser
275 280 285Pro Gly Ile Val Asp Thr Asn
Val Thr Arg Asp Leu Pro Lys Ile Val 290 295
300Val Ala Ala Tyr Arg Phe Leu Pro Tyr Phe Ile Phe Asp Gly Gln
Glu305 310 315 320Gly Ser
Arg Ser Ala Leu Phe Ala Ala Cys Asp Pro Gln Val Pro Glu
325 330 335Tyr Cys Glu Met Leu Lys Ser
Glu Asp Trp Pro Val Cys Ala Cys Ile 340 345
350Asn Tyr Asp Cys Asn Pro Met Asn Ala Ser Glu Glu Ala His
Ser Leu 355 360 365Glu Thr Ser Gln
Leu Val Trp Glu Lys Thr Leu Glu Met Ile Gly Leu 370
375 380Pro Pro Asp Ala Leu Asp Lys Leu Ile Ala Gly Glu
Thr Val Pro Cys385 390 395
400Arg Tyr Gly Gln49942PRTZea mays 49Met Glu Gly Asp Asp Phe Thr Pro Glu
Gly Gly Lys Leu Pro Glu Phe1 5 10
15Lys Leu Asp Ala Arg Gln Ala Gln Gly Phe Ile Ser Phe Phe Lys
Lys 20 25 30Leu Pro Gln Asp
Pro Arg Ala Val Arg Leu Phe Asp Arg Arg Asp Tyr 35
40 45Tyr Thr Ala His Gly Glu Asn Ala Thr Phe Ile Ala
Arg Thr Tyr Tyr 50 55 60His Thr Met
Ser Ala Leu Arg Gln Leu Gly Ser Thr Ser Asp Gly Ile65 70
75 80Leu Ser Ala Ser Val Ser Lys Ala
Met Phe Glu Thr Ile Ala Arg Asn 85 90
95Ile Leu Leu Glu Arg Thr Asp Cys Thr Leu Glu Leu Tyr Glu
Gly Ser 100 105 110Gly Ser Asn
Trp Arg Leu Thr Lys Ser Gly Thr Pro Gly Asn Ile Gly 115
120 125Ser Phe Glu Asp Ile Leu Phe Ala Asn Asn Asp
Met Glu Asp Ser Pro 130 135 140Val Ile
Val Ala Leu Phe Pro Ala Cys Arg Glu Ser Gln Leu Tyr Val145
150 155 160Gly Leu Ser Phe Leu Asp Met
Thr Asn Arg Lys Leu Gly Leu Ala Glu 165
170 175Phe Pro Glu Asp Ser Arg Phe Thr Asn Val Glu Ser
Ala Leu Val Ala 180 185 190Leu
Gly Cys Lys Glu Cys Leu Leu Pro Ala Asp Cys Glu Lys Ser Ile 195
200 205Asp Leu Asn Pro Leu Gln Asp Val Ile
Ser Asn Cys Asn Val Leu Leu 210 215
220Thr Glu Lys Lys Lys Ala Asp Phe Lys Ser Arg Asp Leu Ala Gln Asp225
230 235 240Leu Gly Arg Ile
Ile Arg Gly Ser Val Glu Pro Val Arg Asp Leu Leu 245
250 255Ser Gln Phe Asp Tyr Ala Leu Gly Pro Leu
Gly Ala Leu Leu Ser Tyr 260 265
270Ala Glu Leu Leu Ala Asp Asp Thr Asn Tyr Gly Asn Tyr Thr Ile Glu
275 280 285Lys Tyr Asn Leu Asn Cys Tyr
Met Arg Leu Asp Ser Ala Ala Val Arg 290 295
300Ala Leu Asn Ile Ala Glu Gly Lys Thr Asp Val Asn Lys Asn Phe
Ser305 310 315 320Leu Phe
Gly Leu Met Asn Arg Thr Cys Thr Val Gly Met Gly Lys Arg
325 330 335Leu Leu Asn Arg Trp Leu Lys
Gln Pro Leu Leu Asp Val Asn Glu Ile 340 345
350Asn Asn Arg Leu Asp Met Val Gln Ala Phe Val Glu Asp Pro
Glu Leu 355 360 365Arg Gln Gly Leu
Arg Gln Gln Leu Lys Arg Ile Ser Asp Ile Asp Arg 370
375 380Leu Thr His Ser Leu Arg Lys Lys Ser Ala Asn Leu
Gln Pro Val Val385 390 395
400Lys Leu Tyr Gln Ser Cys Ser Arg Ile Pro Tyr Ile Lys Gly Ile Leu
405 410 415Gln Gln Tyr Asn Gly
Gln Phe Ser Thr Leu Ile Arg Ser Lys Phe Leu 420
425 430Glu Pro Leu Glu Glu Trp Met Ala Lys Asn Arg Phe
Gly Arg Phe Ser 435 440 445Ser Leu
Val Glu Thr Ala Ile Asp Leu Ala Gln Leu Glu Asn Gly Glu 450
455 460Tyr Arg Ile Ser Pro Leu Tyr Ser Ser Asp Leu
Gly Val Leu Lys Asp465 470 475
480Glu Leu Ser Val Val Glu Asn His Ile Asn Asn Leu His Val Asp Thr
485 490 495Ala Ser Asp Leu
Asp Leu Ser Val Asp Lys Gln Leu Lys Leu Glu Lys 500
505 510Gly Ser Leu Gly His Val Phe Arg Met Ser Lys
Lys Glu Glu Gln Lys 515 520 525Val
Arg Lys Lys Leu Thr Gly Ser Tyr Leu Ile Ile Glu Thr Arg Lys 530
535 540Asp Gly Val Lys Phe Thr Asn Ser Lys Leu
Lys Asn Leu Ser Asp Gln545 550 555
560Tyr Gln Ala Leu Phe Gly Glu Tyr Thr Ser Cys Gln Lys Lys Val
Val 565 570 575Gly Asp Val
Val Arg Val Ser Gly Thr Phe Ser Glu Val Phe Glu Asn 580
585 590Phe Ala Ala Val Leu Ser Glu Leu Asp Val
Leu Gln Ser Phe Ala Asp 595 600
605Leu Ala Thr Ser Cys Pro Val Pro Tyr Val Arg Pro Asp Ile Thr Ala 610
615 620Ser Asp Glu Gly Asp Ile Val Leu
Leu Gly Ser Arg His Pro Cys Leu625 630
635 640Glu Ala Gln Asp Gly Val Asn Phe Ile Pro Asn Asp
Cys Thr Leu Val 645 650
655Arg Gly Lys Ser Trp Phe Gln Ile Ile Thr Gly Pro Asn Met Gly Gly
660 665 670Lys Ser Thr Phe Ile Arg
Gln Val Gly Val Asn Val Leu Met Ala Gln 675 680
685Val Gly Ser Phe Val Pro Cys Asp Gln Ala Ser Ile Ser Val
Arg Asp 690 695 700Cys Ile Phe Ala Arg
Val Gly Ala Gly Asp Cys Gln Leu His Gly Val705 710
715 720Ser Thr Phe Met Gln Glu Met Leu Glu Thr
Ala Ser Ile Leu Lys Gly 725 730
735Ala Ser Asp Lys Ser Leu Ile Ile Ile Asp Glu Leu Gly Arg Gly Thr
740 745 750Ser Thr Tyr Asp Gly
Phe Gly Leu Ala Trp Ala Ile Cys Glu His Leu 755
760 765Met Glu Val Thr Arg Ala Pro Thr Leu Phe Ala Thr
His Phe His Glu 770 775 780Leu Thr Ala
Leu Ala His Arg Asn Asp Asp Glu His Gln His Ile Ser785
790 795 800Asp Ile Gly Val Ala Asn Tyr
His Val Gly Ala His Ile Asp Pro Leu 805
810 815Ser Arg Lys Leu Thr Met Leu Tyr Lys Val Glu Pro
Gly Ala Cys Asp 820 825 830Gln
Ser Phe Gly Ile His Val Ala Glu Phe Ala Asn Phe Pro Glu Ala 835
840 845Val Val Ala Leu Ala Lys Ser Lys Ala
Ala Glu Leu Glu Asp Phe Ser 850 855
860Thr Thr Pro Thr Phe Ser Asp Asp Leu Lys Asp Glu Val Gly Ser Lys865
870 875 880Arg Lys Arg Val
Phe Ser Pro Asp Asp Ile Thr Arg Gly Ala Ala Arg 885
890 895Ala Arg Leu Phe Leu Glu Glu Phe Ala Ala
Leu Pro Met Asp Glu Met 900 905
910Asp Gly Ser Lys Ile Leu Glu Met Ala Thr Lys Met Lys Ala Asp Leu
915 920 925Gln Lys Asp Ala Ala Asp Asn
Pro Trp Leu Gln Gln Phe Phe 930 935
94050344PRTArabidopsis thaliana 50Met Glu Trp Ile Arg Gly Glu Thr Ile Gly
Tyr Gly Thr Phe Ser Thr1 5 10
15Val Ser Leu Ala Thr Arg Ser Asn Asn Asp Ser Gly Glu Phe Pro Pro
20 25 30Leu Met Ala Val Lys Ser
Ala Asp Ser Tyr Gly Ala Ala Ser Leu Ala 35 40
45Asn Glu Lys Ser Val Leu Asp Asn Leu Gly Asp Asp Cys Asn
Glu Ile 50 55 60Val Arg Cys Phe Gly
Glu Asp Arg Thr Val Glu Asn Gly Glu Glu Met65 70
75 80His Asn Leu Phe Leu Glu Tyr Ala Ser Arg
Gly Ser Leu Glu Ser Tyr 85 90
95Leu Lys Lys Leu Ala Gly Glu Gly Val Pro Glu Ser Thr Val Arg Arg
100 105 110His Thr Gly Ser Val
Leu Arg Gly Leu Arg His Ile His Ala Asn Gly 115
120 125Phe Ala His Cys Asp Leu Lys Leu Gly Asn Ile Leu
Leu Phe Gly Asp 130 135 140Gly Ala Val
Lys Ile Ala Asp Phe Gly Leu Ala Lys Arg Ile Gly Asp145
150 155 160Leu Thr Ala Leu Asn Tyr Gly
Val Gln Ile Arg Gly Thr Pro Leu Tyr 165
170 175Met Ala Pro Glu Ser Val Asn Asp Asn Glu Tyr Gly
Ser Glu Gly Asp 180 185 190Val
Trp Ala Leu Gly Cys Val Val Val Glu Met Phe Ser Gly Lys Thr 195
200 205Ala Trp Ser Leu Lys Glu Gly Ser Asn
Phe Met Ser Leu Leu Leu Arg 210 215
220Ile Gly Val Gly Asp Glu Val Pro Met Ile Pro Glu Glu Leu Ser Glu225
230 235 240Gln Gly Arg Asp
Phe Leu Ser Lys Cys Phe Val Lys Asp Pro Lys Lys 245
250 255Arg Trp Thr Ala Glu Met Leu Leu Asn His
Pro Phe Val Thr Val Asp 260 265
270Val Asp His Asp Val Leu Val Lys Glu Glu Asp Phe Val Val Asn Met
275 280 285Lys Thr Glu Asp Val Ser Thr
Ser Pro Arg Cys Pro Phe Glu Phe Pro 290 295
300Asp Trp Val Ser Val Ser Ser Gly Ser Gln Thr Ile Asp Ser Pro
Asp305 310 315 320Glu Arg
Val Ala Ser Leu Val Thr Asp Met Ile Pro Asp Trp Ser Val
325 330 335Thr Asn Ser Trp Val Thr Val
Arg 340511187PRTArabidopsis thaliana 51Met Arg Asn His Cys Leu
Glu Leu Ser Ser Asn Cys Ser Ser Ile Phe1 5
10 15Ala Ser Ser Lys Ser Asn Pro Arg Phe Ser Pro Ser
Lys Leu Ser Tyr 20 25 30Ser
Thr Phe Phe Ser Arg Ser Ala Ile Tyr Tyr Arg Ser Lys Pro Lys 35
40 45Gln Ala Ser Ser Ser Ser Ser Phe Ser
Thr Phe Pro Pro Cys Leu Asn 50 55
60Arg Lys Ser Ser Leu Thr His Val Leu Lys Pro Val Ser Glu Leu Ala65
70 75 80Asp Thr Thr Thr Lys
Pro Phe Ser Pro Glu Ile Val Gly Lys Arg Thr 85
90 95Asp Leu Lys Lys Ile Met Ile Leu Gly Ala Gly
Pro Ile Val Ile Gly 100 105
110Gln Ala Cys Glu Phe Asp Tyr Ser Gly Thr Gln Ala Cys Lys Ala Leu
115 120 125Arg Glu Glu Gly Tyr Glu Val
Ile Leu Ile Asn Ser Asn Pro Ala Thr 130 135
140Ile Met Thr Asp Pro Glu Thr Ala Asn Arg Thr Tyr Ile Ala Pro
Met145 150 155 160Thr Pro
Glu Leu Val Glu Gln Val Ile Glu Lys Glu Arg Pro Asp Ala
165 170 175Leu Leu Pro Thr Met Gly Gly
Gln Thr Ala Leu Asn Leu Ala Val Ala 180 185
190Leu Ala Glu Ser Gly Ala Leu Glu Lys Tyr Gly Val Glu Leu
Ile Gly 195 200 205Ala Lys Leu Gly
Ala Ile Lys Lys Ala Glu Asp Arg Glu Leu Phe Lys 210
215 220Asp Ala Met Lys Asn Ile Gly Leu Lys Thr Pro Pro
Ser Gly Ile Gly225 230 235
240Thr Thr Leu Asp Glu Cys Phe Asp Ile Ala Glu Lys Ile Gly Glu Phe
245 250 255Pro Leu Ile Ile Arg
Pro Ala Phe Thr Leu Gly Gly Thr Gly Gly Gly 260
265 270Ile Ala Tyr Asn Lys Glu Glu Phe Glu Ser Ile Cys
Lys Ser Gly Leu 275 280 285Ala Ala
Ser Ala Thr Ser Gln Val Leu Val Glu Lys Ser Leu Leu Gly 290
295 300Trp Lys Glu Tyr Glu Leu Glu Val Met Arg Asp
Leu Ala Asp Asn Val305 310 315
320Val Ile Ile Cys Ser Ile Glu Asn Ile Asp Pro Met Gly Val His Thr
325 330 335Gly Asp Ser Ile
Thr Val Ala Pro Ala Gln Thr Leu Thr Asp Arg Glu 340
345 350Tyr Gln Arg Leu Arg Asp Tyr Ser Ile Ala Ile
Ile Arg Glu Ile Gly 355 360 365Val
Glu Cys Gly Gly Ser Asn Val Gln Phe Ala Val Asn Pro Val Asp 370
375 380Gly Glu Val Met Ile Ile Glu Met Asn Pro
Arg Val Ser Arg Ser Ser385 390 395
400Ala Leu Ala Ser Lys Ala Thr Gly Phe Pro Ile Ala Lys Met Ala
Ala 405 410 415Lys Leu Ser
Val Gly Tyr Thr Leu Asp Gln Ile Pro Asn Asp Ile Thr 420
425 430Arg Lys Thr Pro Ala Ser Phe Glu Pro Ser
Ile Asp Tyr Val Val Thr 435 440
445Lys Ile Pro Arg Phe Ala Phe Glu Lys Phe Pro Gly Ser Gln Pro Leu 450
455 460Leu Thr Thr Gln Met Lys Ser Val
Gly Glu Ser Met Ala Leu Gly Arg465 470
475 480Thr Phe Gln Glu Ser Phe Gln Lys Ala Leu Arg Ser
Leu Glu Cys Gly 485 490
495Phe Ser Gly Trp Gly Cys Ala Lys Ile Lys Glu Leu Asp Trp Asp Trp
500 505 510Asp Gln Leu Lys Tyr Ser
Leu Arg Val Pro Asn Pro Asp Arg Ile His 515 520
525Ala Ile Tyr Ala Ala Met Lys Lys Gly Met Lys Ile Asp Glu
Ile Tyr 530 535 540Glu Leu Ser Met Val
Asp Lys Trp Phe Leu Thr Gln Leu Lys Glu Leu545 550
555 560Val Asp Val Glu Gln Tyr Leu Met Ser Gly
Thr Leu Ser Glu Ile Thr 565 570
575Lys Glu Asp Leu Tyr Glu Val Lys Lys Arg Gly Phe Ser Asp Lys Gln
580 585 590Ile Ala Phe Ala Thr
Lys Thr Thr Glu Glu Glu Val Arg Thr Lys Arg 595
600 605Ile Ser Leu Gly Val Val Pro Ser Tyr Lys Arg Val
Asp Thr Cys Ala 610 615 620Ala Glu Phe
Glu Ala His Thr Pro Tyr Met Tyr Ser Ser Tyr Asp Val625
630 635 640Glu Cys Glu Ser Ala Pro Asn
Asn Lys Lys Lys Val Leu Ile Leu Gly 645
650 655Gly Gly Pro Asn Arg Ile Gly Gln Gly Ile Glu Phe
Asp Tyr Cys Cys 660 665 670Cys
His Thr Ser Phe Ala Leu Gln Asp Ala Gly Tyr Glu Thr Ile Met 675
680 685Leu Asn Ser Asn Pro Glu Thr Val Ser
Thr Asp Tyr Asp Thr Ser Asp 690 695
700Arg Leu Tyr Phe Glu Pro Leu Thr Ile Glu Asp Val Leu Asn Val Ile705
710 715 720Asp Leu Glu Lys
Pro Asp Gly Ile Ile Val Gln Phe Gly Gly Gln Thr 725
730 735Pro Leu Lys Leu Ala Leu Pro Ile Lys His
Tyr Leu Asp Lys His Met 740 745
750Pro Met Ser Leu Ser Gly Ala Gly Pro Val Arg Ile Trp Gly Thr Ser
755 760 765Pro Asp Ser Ile Asp Ala Ala
Glu Asp Arg Glu Arg Phe Asn Ala Ile 770 775
780Leu Asp Glu Leu Lys Ile Glu Gln Pro Lys Gly Gly Ile Ala Lys
Ser785 790 795 800Glu Ala
Asp Ala Leu Ala Ile Ala Lys Glu Val Gly Tyr Pro Val Val
805 810 815Val Arg Pro Ser Tyr Val Leu
Gly Gly Arg Ala Met Glu Ile Val Tyr 820 825
830Asp Asp Ser Arg Leu Ile Thr Tyr Leu Glu Asn Ala Val Gln
Val Asp 835 840 845Pro Glu Arg Pro
Val Leu Val Asp Lys Tyr Leu Ser Asp Ala Ile Glu 850
855 860Ile Asp Val Asp Thr Leu Thr Asp Ser Tyr Gly Asn
Val Val Ile Gly865 870 875
880Gly Ile Met Glu His Ile Glu Gln Ala Gly Val His Ser Gly Asp Ser
885 890 895Ala Cys Met Leu Pro
Thr Gln Thr Ile Pro Ala Ser Cys Leu Gln Thr 900
905 910Ile Arg Thr Trp Thr Thr Lys Leu Ala Lys Lys Leu
Asn Val Cys Gly 915 920 925Leu Met
Asn Cys Gln Tyr Ala Ile Thr Thr Ser Gly Asp Val Phe Leu 930
935 940Leu Glu Ala Asn Pro Arg Ala Ser Arg Thr Val
Pro Phe Val Ser Lys945 950 955
960Ala Ile Gly His Pro Leu Ala Lys Tyr Ala Ala Leu Val Met Ser Gly
965 970 975Lys Ser Leu Lys
Asp Leu Asn Phe Glu Lys Glu Val Ile Pro Lys His 980
985 990Val Ser Val Lys Glu Ala Val Phe Pro Phe Glu
Lys Phe Gln Gly Cys 995 1000
1005Asp Val Ile Leu Gly Pro Glu Met Arg Ser Thr Gly Glu Val Met
1010 1015 1020Ser Ile Ser Ser Glu Phe
Ser Ser Ala Phe Ala Met Ala Gln Ile 1025 1030
1035Ala Ala Gly Gln Lys Leu Pro Leu Ser Gly Thr Val Phe Leu
Ser 1040 1045 1050Leu Asn Asp Met Thr
Lys Pro His Leu Glu Lys Ile Ala Val Ser 1055 1060
1065Phe Leu Glu Leu Gly Phe Lys Ile Val Ala Thr Ser Gly
Thr Ala 1070 1075 1080His Phe Leu Glu
Leu Lys Gly Ile Pro Val Glu Arg Val Leu Lys 1085
1090 1095Leu His Glu Gly Arg Pro His Ala Ala Asp Met
Val Ala Asn Gly 1100 1105 1110Gln Ile
His Leu Met Leu Ile Thr Ser Ser Gly Asp Ala Leu Asp 1115
1120 1125Gln Lys Asp Gly Arg Gln Leu Arg Gln Met
Ala Leu Ala Tyr Lys 1130 1135 1140Val
Pro Val Ile Thr Thr Val Ala Gly Ala Leu Ala Thr Ala Glu 1145
1150 1155Gly Ile Lys Ser Leu Lys Ser Ser Ala
Ile Lys Met Thr Ala Leu 1160 1165
1170Gln Asp Phe Phe Glu Val Lys Asn Val Ser Ser Leu Leu Val 1175
1180 118552274PRTGlycine max 52Met Arg Ala
Lys Leu Phe Val Phe Pro Ile Arg Gly Arg Asn Trp Cys1 5
10 15Phe Ser Arg Thr Ile Asp His Ser Leu
Ser Ala Ser His Ala Ser Ser 20 25
30Gln Ser Pro Ser Thr Leu Lys Asp Leu Trp Thr Asn Ile Asn Val Gly
35 40 45Asp Lys Pro Leu Asn Thr Lys
Thr Glu Leu Phe Val Asp Tyr Ile Ala 50 55
60Asn Lys Met Asn Asn Ala Trp Ile Gly Leu Glu Lys Ala Pro Glu Gly65
70 75 80Ser Phe Lys Asn
Lys Ile His Gly Leu Gly Leu Arg Leu Leu Ser Arg 85
90 95Val Lys Pro Ser Glu Ile Phe Leu Lys Ser
Ile Ser Lys Glu Ile Thr 100 105
110Ser Val Glu Ile Ile Tyr Pro Ser Ser Leu Asn Ala Gln Leu Val Arg
115 120 125Arg Arg Leu Arg His Ile Ala
Val Arg Gly Ala Val Ile His Arg Asn 130 135
140Tyr Leu Tyr Gly Leu Val Ser Leu Ile Pro Leu Thr Ser Ala Leu
Ser145 150 155 160Ile Leu
Pro Leu Pro Asn Val Pro Phe Phe Trp Val Leu Phe Arg Thr
165 170 175Tyr Ser His Trp Arg Ala Leu
Gln Gly Ser Glu Arg Leu Phe Gln Leu 180 185
190Val Ser Asp Asn Ser Lys Thr Ser Asn Thr Cys Thr Tyr Glu
Lys Lys 195 200 205Thr Glu His Lys
Glu Ser Lys Ser Gln Arg His Ser Ser Asn Glu Pro 210
215 220Cys Trp Val Leu Arg Pro Ser Lys Glu Leu Glu Asn
Leu Val His Leu225 230 235
240Glu Asp Gly Gln Glu Ser Leu Ser Gln His Ala Ile Ile Asn Ile Cys
245 250 255Lys Ile Tyr Asp Leu
Asn Pro Val Asp Val Ile Lys Tyr Glu Lys Ser 260
265 270Val Phe53206PRTArabidopsis thaliana 53Met Ala Lys
Glu Ser Thr Thr Ile Asp Val Gly Glu Pro Ser Thr Val1 5
10 15Thr Lys Ser Ser Ser His Val Val Lys
Asp Ala Lys Lys Lys Gly Phe 20 25
30Val Ala Val Ala Ser Arg Gly Gly Ala Lys Arg Gly Leu Ala Ile Phe
35 40 45Asp Phe Leu Leu Arg Leu Ala
Ala Ile Ala Val Thr Ile Gly Ala Ala 50 55
60Ser Val Met Tyr Thr Ala Glu Glu Thr Leu Pro Phe Phe Thr Gln Phe65
70 75 80Leu Gln Phe Gln
Ala Gly Tyr Asp Asp Leu Pro Ala Phe Gln Tyr Phe 85
90 95Val Ile Ala Val Ala Val Val Ala Ser Tyr
Leu Val Leu Ser Leu Pro 100 105
110Phe Ser Ile Val Ser Ile Val Arg Pro His Ala Val Ala Pro Arg Leu
115 120 125Ile Leu Leu Ile Cys Asp Thr
Leu Val Val Thr Leu Asn Thr Ser Ala 130 135
140Ala Ala Ala Ala Ala Ser Ile Thr Tyr Leu Ala His Asn Gly Asn
Gln145 150 155 160Ser Thr
Asn Trp Leu Pro Ile Cys Gln Gln Phe Gly Asp Phe Cys Gln
165 170 175Asn Val Ser Thr Ala Val Val
Ala Asp Ser Ile Ala Ile Leu Phe Phe 180 185
190Ile Val Leu Ile Ile Ile Ser Ala Ile Ala Leu Lys Arg His
195 200 20554411PRTEscherichia coli
54Met Ala Thr Ala Thr Ser Ala Ser Leu Phe Ser Thr Val Ser Ser Ser1
5 10 15Tyr Ser Lys Ala Ser Ser
Ile Pro His Ser Arg Leu Gln Ser Val Lys 20 25
30Phe Asn Ser Val Pro Ser Phe Thr Gly Leu Lys Ser Thr
Ser Leu Ile 35 40 45Ser Gly Ser
Asp Ser Ser Ser Leu Ala Lys Thr Leu Arg Gly Ser Val 50
55 60Thr Lys Ala Gln Thr Ser Asp Lys Lys Pro Tyr Gly
Phe Lys Ile Asn65 70 75
80Ala Met Lys Thr Ala Tyr Ile Ala Lys Gln Arg Gln Ile Ser Phe Val
85 90 95Lys Ser His Phe Ser Arg
Gln Leu Glu Glu Arg Leu Gly Leu Ile Glu 100
105 110Val Gln Ala Pro Ile Leu Ser Arg Val Gly Asp Gly
Thr Gln Asp Asn 115 120 125Leu Ser
Gly Cys Glu Lys Ala Val Gln Val Lys Val Lys Ala Leu Pro 130
135 140Asp Ala Gln Phe Glu Val Val His Ser Leu Ala
Lys Trp Lys Arg Gln145 150 155
160Thr Leu Gly Gln His Asp Phe Ser Ala Gly Glu Gly Leu Tyr Thr His
165 170 175Met Lys Ala Leu
Arg Pro Asp Glu Asp Arg Leu Ser Pro Leu His Ser 180
185 190Val Tyr Val Asp Gln Trp Asp Trp Glu Arg Val
Met Gly Asp Gly Glu 195 200 205Arg
Gln Phe Ser Thr Leu Lys Ser Thr Val Glu Ala Ile Trp Ala Gly 210
215 220Ile Lys Ala Thr Glu Ala Ala Val Ser Glu
Glu Phe Gly Leu Ala Pro225 230 235
240Phe Leu Pro Asp Gln Ile His Phe Val His Ser Gln Glu Leu Leu
Ser 245 250 255Arg Tyr Pro
Asp Leu Asp Ala Lys Gly Arg Glu Arg Ala Ile Ala Lys 260
265 270Asp Leu Gly Ala Val Phe Leu Val Gly Ile
Gly Gly Lys Leu Ser Asp 275 280
285Gly His Arg His Asp Val Arg Ala Pro Asp Tyr Asp Asp Trp Ser Thr 290
295 300Pro Ser Glu Leu Gly His Ala Gly
Leu Asn Gly Asp Ile Leu Val Trp305 310
315 320Asn Pro Val Leu Glu Asp Ala Phe Glu Leu Ser Ser
Met Gly Ile Arg 325 330
335Val Asp Ala Asp Thr Leu Lys His Gln Leu Ala Leu Thr Gly Asp Glu
340 345 350Asp Arg Leu Glu Leu Glu
Trp His Gln Ala Leu Leu Arg Gly Glu Met 355 360
365Pro Gln Thr Ile Gly Gly Gly Ile Gly Gln Ser Arg Leu Thr
Met Leu 370 375 380Leu Leu Gln Leu Pro
His Ile Gly Gln Val Gln Cys Gly Val Trp Pro385 390
395 400Ala Ala Val Arg Glu Ser Val Pro Ser Leu
Leu 405 410551173DNAZea mays 55atgcagagcg
cggctgccat cgggctccta cggccatgtg ccgcgcggcc gctcgccgcc 60tacactagcc
cacgccgcgg cgccggcgcg tgcagcggcg gcacccagcc gctcatcacg 120ccccgcggca
tccgcctctc cgcccgcccc ggtctcgtgc cggcctcgcc gctggaggag 180aaggagaacc
ggagatgcag ggccagtatg cacgcggcgg cgtcggccgg agaggaagct 240gggggagggc
tcgccaagac gctgcagctg ggggcgcttt tcgggctctg gtacctcttc 300aacatctact
tcaacatcta caacaagcag gttctgaagg ttttgccata ccctataaac 360atcacaacgg
tgcagtttgc tgttggaagt gccattgctt tgttcatgtg gatcactggt 420atccataaaa
ggccaaagat ttcgggtgcc cagcttttcg ctatccttcc tctagctatt 480gtccatacca
tgggcaatct tttcacaaac atgagccttg gaaaggttgc agtgtcattt 540acacatacta
taaaggccat ggaacctttc ttctcagttc tcctttcagc aattttcctt 600ggggagttgc
ctacgccatg ggttgtgttg tctcttcttc cgattgttgg tggtgtagct 660ttggcatccc
ttactgaggc ctcctttaac tgggctggat tttggagtgc aatggcttca 720aatgtaacct
tccagtcaag gaatgtgcta agcaagaaac ttatggtgaa gaaagaggaa 780tctctcgaca
acattaacct attctcgatc attacagtca tgtcattctt cctgttggcc 840ccagtaacct
tacttacaga aggtgttaaa gttagtccag cagtgttgca gtctgctggt 900ttgaacttga
aacaggtata cacaaggtca ttgattgctg cattctgctt ccatgcatac 960caacaggtat
catacatgat cctcgccagg gtatccccag tcacacattc agtgggcaat 1020tgcgtcaagc
gtgtggtggt cattgtgacc tctgttctgt tcttcaggac ccctgtttct 1080cccatcaact
ctcttggtac cgggatcgct cttgctggag ttttcctata ctcgcaattg 1140aagagactta
agcccaagcc caagactgct tga
117356720DNAGlycine max 56atggctgctg ctagtactat gatcacactc aagttctcat
tcctctggag tctccttctc 60tcagtttcac tcttgggtgt aaagtcatca catagccacc
aaaatggtgg gagaagaaca 120attcctccaa catgcaagcg cattgagtgc cccacccatg
atgtgattga agtgggtgat 180ggctatgaaa tccgacgcta taataataat tcaactgtgt
ggatgtcaac ttctcccatt 240caagacattt ctctggttga agctacaaga actggcttca
ggagtctatt tgattatatc 300caaggcaaga acaactacaa gcaaaaaatt gagatgacag
cgcctgtgat cacagaagtt 360tcacctagtg atggaccctt ttgtaaatcc tcatttgttg
tcagcttctt tgtgccaaaa 420ttgaaccaag caaaccctcc tcctgcaaag ggtctccatg
tccaaagatg gaacaatatg 480tatgtggcag caaggcagtt tggtggacac gtaaacgatt
caaatgttgc ggtggaagcc 540gctgtgttgc gagctagtat tgaaggcaca aaatggtctg
gtgccattga caaaaaccag 600aaagctggcc atgcttctgt ttacactgtg gcacaataca
atgacccttt tgaatatcag 660aatagggtga atgagatatg gttcttgttt gaaatggaaa
gtgaaaggca tgcaatttga 72057981DNAGlycine max 57atggaacgaa gtggcggaat
ggtaacgggg tcgcatgaaa ggaacgaact tgttagagtt 60agacacggtt ctgacagtgg
gtctaaaccc ttgaagaatt taaatggtca gatttgtcaa 120atatgtggtg acaccattgg
attaacggct actggtgacc tctttgttgc ttgtcatgag 180tgtggcttcc cactttgtca
ttcttgttac gagtatgagc tgaaaaatgt gagccaatct 240tgtccccagt gcaagactac
attcacaagt cgccaagagg gtgctgaagt ggagggagat 300gatgatgacg aagacgatgc
tgatgatcta gataatggga tcaactatgg ccaaggaaac 360aattccaagt cggggatgct
gtgggaagaa gatgctgacc tctcttcatc ttctggacat 420gattctcata taccaaaccc
ccatctagta aacgggcaac cgatgtctgg tgagtttcca 480tgtgctactt ctgatgctca
atctatgcaa actacatcag atcctatggg tcaatccgaa 540aaggttcact cacttccata
tgctgatcca aagcaaccag gtcctgagag tgatgaagag 600ataagaagag tgccggagat
tggaggtgaa agcgctggaa cttcagcctc tcggccagat 660gccggttcaa atgctggtac
agaacgtgct caggggacag gggacagcca gaagaagaga 720gggagaagcc cagctgataa
agaaagcaag cggctaaaga ggctactgag gaatagagtt 780tcggctcagc aagcaaggga
gaggaagaag gcatatttga ttgatttgga aacaagagtc 840aaagacttag agaagaagaa
ctcagagctc aaagaaagac tttccacttt gcagaatgaa 900aaccaaatgc ttagacaaat
attgaagaac acaacagcaa gcaggagagg gagcaatagt 960ggtaccaata atgctgagta a
98158792DNAZea mays
58atgttggagc tacgtctcgt gcagggctct ctcctgaaga aggttctcga atcgatcaag
60gatctcgtca acgacgccaa cttcgactgc tccaccaccg gcttctccct ccaagccatg
120gactccagcc acgtggcgct cgtctccctc ctgctcagat ccgaaggctt cgagcactac
180cgctgcgacc gtaacctttc catggggatg aatctcggaa acatgtcgaa gatgctcaaa
240tgcgccggaa acgacgacat catcaccatc aaagccgatg acggcggcga caccgtcacc
300ttcatgttcg agagtcccaa gcaagacaag attgcagatt ttgagatgaa gctgatggat
360atagacagtg agcatttggg gatacctgat gctgagtatc attcgattgt taggatgccg
420tctaatgagt tctctagaat ctgcaaagat ctcagtacca tcggtgacac tgttgtgata
480tctgtgacta aagaaggggt taagttctct actgctggtg acattgggac agctaacatt
540gtgttgagac agaacacaac tgttgacaag ccggaagatg cgattgtaat agagatgaac
600gagccggtgt cactctcgtt tgccttgagg tatatgaatt ccttcacaaa ggcgactcct
660ttgtcagaca cggtgacgat cagcttatcg tcggagctgc cagttgtggt ggagtataag
720gtggctgaga tgggttacat tcgttactac ttggctccta agattgaaga agatgaagaa
780gacaaggctt aa
79259902DNAZea mays 59atggcggcgc cgcgcgtcct cctcctcctc gccgccgcgg
cccttctcgc cgtcgcctcc 60ctcggggacg cttcgggcga gggcccccgc gggcgcaagc
tgctggtgct cgtcgacgat 120ctggccgtcc gctcatccca ctcggccttc ttcggctcgc
tccaggcccg cgggctagat 180ctggagttcc gcctcgccga cgaccccaag ctctcgctcc
accgctacgg tcagtacctc 240tacgacggcc tcgtgctctt cgccccatcg accccgcgct
tcggcggatc ggtggaccag 300aatgctgttc tggagtttat cgatgccggg catgatatga
ttctggcagc agatcattcg 360gcttcagatc tgatccgcgg catcgcgacg gagtgtgggg
ttgactttga tgaggacccg 420gaagcaatgg ttattgacca catcaattat gcctccagtg
aggttgaagg tgaccacacc 480ttgattgctg gcgatgacct gattcagtca gatgtgatat
tggggtccaa aaagattgag 540gctcctgtgc tgtttcgagg gattgggcat gcggccaatc
catccaacag cttggtttta 600aaggttctat ctgcctcgcc atcagcgtat tcagcaaacc
cggaggctaa gtggcatctg 660ttccatctct cactgggtcg gccatatcgc tggtttctgt
tatgcaggct aggaataatg 720ctcgtgtgtt gatatctgga tcactggatt tgtttagcaa
caggttccta aagtctggtg 780tgcagaaggc tggcagcaaa atgagccatg acaaagctgg
aaatgaacaa tttgtgacag 840agacgagcaa atgggtcttc catgagaggg ggcatctgaa
ggcagggaat gtcaagcacc 900at
902602184DNAZea mays 60atggccgcgg ggtcgatccg
ggtcaccatg gaggtgggcg ccgacggcgt cgcgctcatc 60accatcgcca acccgcccgt
caacgcgctc caccccatca tcatcgcggg gctcaaggac 120aagtacgcgg aggccttgcg
ccgtgacgac gttaaggcaa tcgtgctcac tggtgctgga 180ggcaagttct gtggaggatt
tgatatcaac gttttcacaa aggttcatca gactggggat 240gtatcactta tgccggacgt
atccgtcgag cttgtgtcaa acatgatgga agagggaaaa 300aaaccttctg ttgcagccat
tcaaggtctt gcattgggtg gtggcctaga gttgactatg 360ggttgtcatg ctcggatatc
tactcctgaa gctcaacttg gattgccaga gctaaccctt 420ggcatcatcc ctggatttgg
aggtacccag cgtttgccga ggcttgtagg tctacccaaa 480gcaattgaaa tgatgctgca
aagtaagttc attacggcaa aggaagggaa tgaacgtggt 540ttgattgatg ccctttgctc
tcctgatgaa ttgataaaga catcacgtct ttgggctctg 600gaaattgcta attgccgtaa
accttggatg aggtctcttg gcagaacaga taggcttgga 660ccactctctg aagctcgtgc
tgtgttaaat gcagcaagac agcaagcaat gaagatcgca 720ccaaacatgc cacaaaacca
ggcctgcctg gatgtgatgg aggaaggcat attatgtgga 780ggccaagctg gtgttttgaa
ggaggccgtg gttttcaagg agctggtgat agcaccaaca 840tcaaaggctc ttgtccatgt
tttctttgca caacgttcca cgacaaaggt gccaggtgta 900actgatgttc aactgaaacc
aaggccaatt agaaaagttg ctgttattgg tggtggtctg 960atgggatctg gaattgccac
atcacttctt gttagcaaca tttctgttgt gctcaaggaa 1020gtaaaccctc agtttctgca
aaggggagag aaaacaatag caggtaatct tgagggcctg 1080gtcaaaagag gttcactaac
aaaggatagg atgcacaagg ccatggccct tctcaagggt 1140gctttggatt attcagattt
caaggatgtt gatatggtta ttgaggctgt tattgagaag 1200attcctttga agcaatcaat
atttgctgac attgagaaaa tctgtccaaa acattgcata 1260cttgcaacaa acacatccac
cattgatttg aatgttgttg gcaagaagac aaattctcaa 1320gatagaatta taggggctca
ctttttcagc cctgctcata ttatgccctt gcttgaaatt 1380gttcggacgg agaagacatc
accacaagct atccttgatc tcatcaccat tgggaagata 1440ataaagaaag tccctattgt
ggtcggcaac tgcacaggat ttgcagtcaa ccgtacattt 1500tttccttaca cacagggttc
tcatcttcta gttagtcttg gtattgatgt tttcagaatt 1560gatcgagtaa taagcacctt
tggcatgcca atgggacctt ttcaactcca agatgtggct 1620gggtatggag ttgccttggc
agtaaaggat atctacgctg atgcctttgg agaaagaaat 1680ttggactctg accttgtgga
tttgatggta aaggatggac gacaaggaaa ggtgaacggc 1740aaaggttact acatttatga
gaagggtggg aagccaaagc cagatcctag tgttaagcat 1800gttatcgagg agtaccgaaa
gcacgcaaac acaatgcctg gtggaaagcc tgttacttta 1860acggatcaag atattttgga
gatgattttc ttcccagttg tgaatgaggc atgcagggtt 1920atggatgaaa atgttgtaat
tcgagcttct gatcttgata ttgcttctgt tcttggaatg 1980ggctttccta aatacagggg
tggtcttgtc ttctgggctg acactgttgg agcaccttac 2040atacattcta agctaagcaa
gtgggctgaa atttatggcc ccttcttcaa accatcatca 2100tatttggaac agcgagctaa
gagtggtgta ccattgagcg caccaggagc ttcgcagcaa 2160ggttcggcga ggtcacgcat
gtga 218461390PRTZea mays 61Met
Gln Ser Ala Ala Ala Ile Gly Leu Leu Arg Pro Cys Ala Ala Arg1
5 10 15Pro Leu Ala Ala Tyr Thr Ser
Pro Arg Arg Gly Ala Gly Ala Cys Ser 20 25
30Gly Gly Thr Gln Pro Leu Ile Thr Pro Arg Gly Ile Arg Leu
Ser Ala 35 40 45Arg Pro Gly Leu
Val Pro Ala Ser Pro Leu Glu Glu Lys Glu Asn Arg 50 55
60Arg Cys Arg Ala Ser Met His Ala Ala Ala Ser Ala Gly
Glu Glu Ala65 70 75
80Gly Gly Gly Leu Ala Lys Thr Leu Gln Leu Gly Ala Leu Phe Gly Leu
85 90 95Trp Tyr Leu Phe Asn Ile
Tyr Phe Asn Ile Tyr Asn Lys Gln Val Leu 100
105 110Lys Val Leu Pro Tyr Pro Ile Asn Ile Thr Thr Val
Gln Phe Ala Val 115 120 125Gly Ser
Ala Ile Ala Leu Phe Met Trp Ile Thr Gly Ile His Lys Arg 130
135 140Pro Lys Ile Ser Gly Ala Gln Leu Phe Ala Ile
Leu Pro Leu Ala Ile145 150 155
160Val His Thr Met Gly Asn Leu Phe Thr Asn Met Ser Leu Gly Lys Val
165 170 175Ala Val Ser Phe
Thr His Thr Ile Lys Ala Met Glu Pro Phe Phe Ser 180
185 190Val Leu Leu Ser Ala Ile Phe Leu Gly Glu Leu
Pro Thr Pro Trp Val 195 200 205Val
Leu Ser Leu Leu Pro Ile Val Gly Gly Val Ala Leu Ala Ser Leu 210
215 220Thr Glu Ala Ser Phe Asn Trp Ala Gly Phe
Trp Ser Ala Met Ala Ser225 230 235
240Asn Val Thr Phe Gln Ser Arg Asn Val Leu Ser Lys Lys Leu Met
Val 245 250 255Lys Lys Glu
Glu Ser Leu Asp Asn Ile Asn Leu Phe Ser Ile Ile Thr 260
265 270Val Met Ser Phe Phe Leu Leu Ala Pro Val
Thr Leu Leu Thr Glu Gly 275 280
285Val Lys Val Ser Pro Ala Val Leu Gln Ser Ala Gly Leu Asn Leu Lys 290
295 300Gln Val Tyr Thr Arg Ser Leu Ile
Ala Ala Phe Cys Phe His Ala Tyr305 310
315 320Gln Gln Val Ser Tyr Met Ile Leu Ala Arg Val Ser
Pro Val Thr His 325 330
335Ser Val Gly Asn Cys Val Lys Arg Val Val Val Ile Val Thr Ser Val
340 345 350Leu Phe Phe Arg Thr Pro
Val Ser Pro Ile Asn Ser Leu Gly Thr Gly 355 360
365Ile Ala Leu Ala Gly Val Phe Leu Tyr Ser Gln Leu Lys Arg
Leu Lys 370 375 380Pro Lys Pro Lys Thr
Ala385 39062239PRTGlycine max 62Met Ala Ala Ala Ser Thr
Met Ile Thr Leu Lys Phe Ser Phe Leu Trp1 5
10 15Ser Leu Leu Leu Ser Val Ser Leu Leu Gly Val Lys
Ser Ser His Ser 20 25 30His
Gln Asn Gly Gly Arg Arg Thr Ile Pro Pro Thr Cys Lys Arg Ile 35
40 45Glu Cys Pro Thr His Asp Val Ile Glu
Val Gly Asp Gly Tyr Glu Ile 50 55
60Arg Arg Tyr Asn Asn Asn Ser Thr Val Trp Met Ser Thr Ser Pro Ile65
70 75 80Gln Asp Ile Ser Leu
Val Glu Ala Thr Arg Thr Gly Phe Arg Ser Leu 85
90 95Phe Asp Tyr Ile Gln Gly Lys Asn Asn Tyr Lys
Gln Lys Ile Glu Met 100 105
110Thr Ala Pro Val Ile Thr Glu Val Ser Pro Ser Asp Gly Pro Phe Cys
115 120 125Lys Ser Ser Phe Val Val Ser
Phe Phe Val Pro Lys Leu Asn Gln Ala 130 135
140Asn Pro Pro Pro Ala Lys Gly Leu His Val Gln Arg Trp Asn Asn
Met145 150 155 160Tyr Val
Ala Ala Arg Gln Phe Gly Gly His Val Asn Asp Ser Asn Val
165 170 175Ala Val Glu Ala Ala Val Leu
Arg Ala Ser Ile Glu Gly Thr Lys Trp 180 185
190Ser Gly Ala Ile Asp Lys Asn Gln Lys Ala Gly His Ala Ser
Val Tyr 195 200 205Thr Val Ala Gln
Tyr Asn Asp Pro Phe Glu Tyr Gln Asn Arg Val Asn 210
215 220Glu Ile Trp Phe Leu Phe Glu Met Glu Ser Glu Arg
His Ala Ile225 230 23563326PRTGlycine max
63Met Glu Arg Ser Gly Gly Met Val Thr Gly Ser His Glu Arg Asn Glu1
5 10 15Leu Val Arg Val Arg His
Gly Ser Asp Ser Gly Ser Lys Pro Leu Lys 20 25
30Asn Leu Asn Gly Gln Ile Cys Gln Ile Cys Gly Asp Thr
Ile Gly Leu 35 40 45Thr Ala Thr
Gly Asp Leu Phe Val Ala Cys His Glu Cys Gly Phe Pro 50
55 60Leu Cys His Ser Cys Tyr Glu Tyr Glu Leu Lys Asn
Val Ser Gln Ser65 70 75
80Cys Pro Gln Cys Lys Thr Thr Phe Thr Ser Arg Gln Glu Gly Ala Glu
85 90 95Val Glu Gly Asp Asp Asp
Asp Glu Asp Asp Ala Asp Asp Leu Asp Asn 100
105 110Gly Ile Asn Tyr Gly Gln Gly Asn Asn Ser Lys Ser
Gly Met Leu Trp 115 120 125Glu Glu
Asp Ala Asp Leu Ser Ser Ser Ser Gly His Asp Ser His Ile 130
135 140Pro Asn Pro His Leu Val Asn Gly Gln Pro Met
Ser Gly Glu Phe Pro145 150 155
160Cys Ala Thr Ser Asp Ala Gln Ser Met Gln Thr Thr Ser Asp Pro Met
165 170 175Gly Gln Ser Glu
Lys Val His Ser Leu Pro Tyr Ala Asp Pro Lys Gln 180
185 190Pro Gly Pro Glu Ser Asp Glu Glu Ile Arg Arg
Val Pro Glu Ile Gly 195 200 205Gly
Glu Ser Ala Gly Thr Ser Ala Ser Arg Pro Asp Ala Gly Ser Asn 210
215 220Ala Gly Thr Glu Arg Ala Gln Gly Thr Gly
Asp Ser Gln Lys Lys Arg225 230 235
240Gly Arg Ser Pro Ala Asp Lys Glu Ser Lys Arg Leu Lys Arg Leu
Leu 245 250 255Arg Asn Arg
Val Ser Ala Gln Gln Ala Arg Glu Arg Lys Lys Ala Tyr 260
265 270Leu Ile Asp Leu Glu Thr Arg Val Lys Asp
Leu Glu Lys Lys Asn Ser 275 280
285Glu Leu Lys Glu Arg Leu Ser Thr Leu Gln Asn Glu Asn Gln Met Leu 290
295 300Arg Gln Ile Leu Lys Asn Thr Thr
Ala Ser Arg Arg Gly Ser Asn Ser305 310
315 320Gly Thr Asn Asn Ala Glu
32564263PRTZea mays 64Met Leu Glu Leu Arg Leu Val Gln Gly Ser Leu Leu Lys
Lys Val Leu1 5 10 15Glu
Ser Ile Lys Asp Leu Val Asn Asp Ala Asn Phe Asp Cys Ser Thr 20
25 30Thr Gly Phe Ser Leu Gln Ala Met
Asp Ser Ser His Val Ala Leu Val 35 40
45Ser Leu Leu Leu Arg Ser Glu Gly Phe Glu His Tyr Arg Cys Asp Arg
50 55 60Asn Leu Ser Met Gly Met Asn Leu
Gly Asn Met Ser Lys Met Leu Lys65 70 75
80Cys Ala Gly Asn Asp Asp Ile Ile Thr Ile Lys Ala Asp
Asp Gly Gly 85 90 95Asp
Thr Val Thr Phe Met Phe Glu Ser Pro Lys Gln Asp Lys Ile Ala
100 105 110Asp Phe Glu Met Lys Leu Met
Asp Ile Asp Ser Glu His Leu Gly Ile 115 120
125Pro Asp Ala Glu Tyr His Ser Ile Val Arg Met Pro Ser Asn Glu
Phe 130 135 140Ser Arg Ile Cys Lys Asp
Leu Ser Thr Ile Gly Asp Thr Val Val Ile145 150
155 160Ser Val Thr Lys Glu Gly Val Lys Phe Ser Thr
Ala Gly Asp Ile Gly 165 170
175Thr Ala Asn Ile Val Leu Arg Gln Asn Thr Thr Val Asp Lys Pro Glu
180 185 190Asp Ala Ile Val Ile Glu
Met Asn Glu Pro Val Ser Leu Ser Phe Ala 195 200
205Leu Arg Tyr Met Asn Ser Phe Thr Lys Ala Thr Pro Leu Ser
Asp Thr 210 215 220Val Thr Ile Ser Leu
Ser Ser Glu Leu Pro Val Val Val Glu Tyr Lys225 230
235 240Val Ala Glu Met Gly Tyr Ile Arg Tyr Tyr
Leu Ala Pro Lys Ile Glu 245 250
255Glu Asp Glu Glu Asp Lys Ala 26065243PRTZea mays 65Met
Ala Ala Pro Arg Val Leu Leu Leu Leu Ala Ala Ala Ala Leu Leu1
5 10 15Ala Val Ala Ser Leu Gly Asp
Ala Ser Gly Glu Gly Pro Arg Gly Arg 20 25
30Lys Leu Leu Val Leu Val Asp Asp Leu Ala Val Arg Ser Ser
His Ser 35 40 45Ala Phe Phe Gly
Ser Leu Gln Ala Arg Gly Leu Asp Leu Glu Phe Arg 50 55
60Leu Ala Asp Asp Pro Lys Leu Ser Leu His Arg Tyr Gly
Gln Tyr Leu65 70 75
80Tyr Asp Gly Leu Val Leu Phe Ala Pro Ser Thr Pro Arg Phe Gly Gly
85 90 95Ser Val Asp Gln Asn Ala
Val Leu Glu Phe Ile Asp Ala Gly His Asp 100
105 110Met Ile Leu Ala Ala Asp His Ser Ala Ser Asp Leu
Ile Arg Gly Ile 115 120 125Ala Thr
Glu Cys Gly Val Asp Phe Asp Glu Asp Pro Glu Ala Met Val 130
135 140Ile Asp His Ile Asn Tyr Ala Ser Ser Glu Val
Glu Gly Asp His Thr145 150 155
160Leu Ile Ala Gly Asp Asp Leu Ile Gln Ser Asp Val Ile Leu Gly Ser
165 170 175Lys Lys Ile Glu
Ala Pro Val Leu Phe Arg Gly Ile Gly His Ala Ala 180
185 190Asn Pro Ser Asn Ser Leu Val Leu Lys Val Leu
Ser Ala Ser Pro Ser 195 200 205Ala
Tyr Ser Ala Asn Pro Glu Ala Lys Trp His Leu Phe His Leu Ser 210
215 220Leu Gly Arg Pro Tyr Arg Trp Phe Leu Leu
Cys Arg Leu Gly Ile Met225 230 235
240Leu Val Cys66727PRTZea mays 66Met Ala Ala Gly Ser Ile Arg Val
Thr Met Glu Val Gly Ala Asp Gly1 5 10
15Val Ala Leu Ile Thr Ile Ala Asn Pro Pro Val Asn Ala Leu
His Pro 20 25 30Ile Ile Ile
Ala Gly Leu Lys Asp Lys Tyr Ala Glu Ala Leu Arg Arg 35
40 45Asp Asp Val Lys Ala Ile Val Leu Thr Gly Ala
Gly Gly Lys Phe Cys 50 55 60Gly Gly
Phe Asp Ile Asn Val Phe Thr Lys Val His Gln Thr Gly Asp65
70 75 80Val Ser Leu Met Pro Asp Val
Ser Val Glu Leu Val Ser Asn Met Met 85 90
95Glu Glu Gly Lys Lys Pro Ser Val Ala Ala Ile Gln Gly
Leu Ala Leu 100 105 110Gly Gly
Gly Leu Glu Leu Thr Met Gly Cys His Ala Arg Ile Ser Thr 115
120 125Pro Glu Ala Gln Leu Gly Leu Pro Glu Leu
Thr Leu Gly Ile Ile Pro 130 135 140Gly
Phe Gly Gly Thr Gln Arg Leu Pro Arg Leu Val Gly Leu Pro Lys145
150 155 160Ala Ile Glu Met Met Leu
Gln Ser Lys Phe Ile Thr Ala Lys Glu Gly 165
170 175Asn Glu Arg Gly Leu Ile Asp Ala Leu Cys Ser Pro
Asp Glu Leu Ile 180 185 190Lys
Thr Ser Arg Leu Trp Ala Leu Glu Ile Ala Asn Cys Arg Lys Pro 195
200 205Trp Met Arg Ser Leu Gly Arg Thr Asp
Arg Leu Gly Pro Leu Ser Glu 210 215
220Ala Arg Ala Val Leu Asn Ala Ala Arg Gln Gln Ala Met Lys Ile Ala225
230 235 240Pro Asn Met Pro
Gln Asn Gln Ala Cys Leu Asp Val Met Glu Glu Gly 245
250 255Ile Leu Cys Gly Gly Gln Ala Gly Val Leu
Lys Glu Ala Val Val Phe 260 265
270Lys Glu Leu Val Ile Ala Pro Thr Ser Lys Ala Leu Val His Val Phe
275 280 285Phe Ala Gln Arg Ser Thr Thr
Lys Val Pro Gly Val Thr Asp Val Gln 290 295
300Leu Lys Pro Arg Pro Ile Arg Lys Val Ala Val Ile Gly Gly Gly
Leu305 310 315 320Met Gly
Ser Gly Ile Ala Thr Ser Leu Leu Val Ser Asn Ile Ser Val
325 330 335Val Leu Lys Glu Val Asn Pro
Gln Phe Leu Gln Arg Gly Glu Lys Thr 340 345
350Ile Ala Gly Asn Leu Glu Gly Leu Val Lys Arg Gly Ser Leu
Thr Lys 355 360 365Asp Arg Met His
Lys Ala Met Ala Leu Leu Lys Gly Ala Leu Asp Tyr 370
375 380Ser Asp Phe Lys Asp Val Asp Met Val Ile Glu Ala
Val Ile Glu Lys385 390 395
400Ile Pro Leu Lys Gln Ser Ile Phe Ala Asp Ile Glu Lys Ile Cys Pro
405 410 415Lys His Cys Ile Leu
Ala Thr Asn Thr Ser Thr Ile Asp Leu Asn Val 420
425 430Val Gly Lys Lys Thr Asn Ser Gln Asp Arg Ile Ile
Gly Ala His Phe 435 440 445Phe Ser
Pro Ala His Ile Met Pro Leu Leu Glu Ile Val Arg Thr Glu 450
455 460Lys Thr Ser Pro Gln Ala Ile Leu Asp Leu Ile
Thr Ile Gly Lys Ile465 470 475
480Ile Lys Lys Val Pro Ile Val Val Gly Asn Cys Thr Gly Phe Ala Val
485 490 495Asn Arg Thr Phe
Phe Pro Tyr Thr Gln Gly Ser His Leu Leu Val Ser 500
505 510Leu Gly Ile Asp Val Phe Arg Ile Asp Arg Val
Ile Ser Thr Phe Gly 515 520 525Met
Pro Met Gly Pro Phe Gln Leu Gln Asp Val Ala Gly Tyr Gly Val 530
535 540Ala Leu Ala Val Lys Asp Ile Tyr Ala Asp
Ala Phe Gly Glu Arg Asn545 550 555
560Leu Asp Ser Asp Leu Val Asp Leu Met Val Lys Asp Gly Arg Gln
Gly 565 570 575Lys Val Asn
Gly Lys Gly Tyr Tyr Ile Tyr Glu Lys Gly Gly Lys Pro 580
585 590Lys Pro Asp Pro Ser Val Lys His Val Ile
Glu Glu Tyr Arg Lys His 595 600
605Ala Asn Thr Met Pro Gly Gly Lys Pro Val Thr Leu Thr Asp Gln Asp 610
615 620Ile Leu Glu Met Ile Phe Phe Pro
Val Val Asn Glu Ala Cys Arg Val625 630
635 640Met Asp Glu Asn Val Val Ile Arg Ala Ser Asp Leu
Asp Ile Ala Ser 645 650
655Val Leu Gly Met Gly Phe Pro Lys Tyr Arg Gly Gly Leu Val Phe Trp
660 665 670Ala Asp Thr Val Gly Ala
Pro Tyr Ile His Ser Lys Leu Ser Lys Trp 675 680
685Ala Glu Ile Tyr Gly Pro Phe Phe Lys Pro Ser Ser Tyr Leu
Glu Gln 690 695 700Arg Ala Lys Ser Gly
Val Pro Leu Ser Ala Pro Gly Ala Ser Gln Gln705 710
715 720Gly Ser Ala Arg Ser Arg Met
72567328DNAArtificialEngineered miRNA precursor 67ggcagagccg tgcccgtctc
atcccctgcc cgtgcaagca gctaggtagg acgatttgag 60cgtggtgtta ggccgaaccg
ctgaaggaag attgctccac tgttgactgc attagatgtt 120gaagaggtac caaaaatgta
ttgcttatat tcagcaatat aatgttcttg gtacctaggc 180aacatctaat atagtcgata
gtggaagaac ggtaacatat gtggtttgca gcaggtgagc 240aggatgggtg tggatgattg
aatatctctg ttcagtgttt tcatcatctg actgaacact 300gaatcagctt gctgacgtta
gaggttag
32868328DNAArtificialEngineered miRNA precursor 68ggcagagccg tgcccgtctc
atcccctgcc cgtgcaagca gctaggtagg acgatttgag 60cgtggtgtta ggccgaaccg
ctgaaggaag attgctccac tgttgactgc atttcatagc 120catcacccac ttcaaatgta
ttgcttatat tcagcaatat aatgttcgaa gtgggttcgg 180gctatgaaat atagtcgata
gtggaagaac ggtaacatat gtggtttgca gcaggtgagc 240aggatgggtg tggatgattg
aatatctctg ttcagtgttt tcatcatctg actgaacact 300gaatcagctt gctgacgtta
gaggttag
32869325DNAArtificialEngineered miRNA precursor 69ggcagagccg tgcccgtctc
atcccctgcc cgtgcaagca gctaggtagg acgatttgag 60cgtggtgtta ggccgaaccg
ctgaaggaag attgctccac tgttgactgc attgacaagc 120aacaaagagg tcaaaatgta
ttgcttatat tcagcaatat aatgttctga cctcttgtgt 180gcttgtcaat atagtcgata
gtggaagaac ggtaacatat gtggtttgca gcaggtgagc 240aggatgggtg tggatgattg
aatatctctg ttcagtgttt tcatcatctg actgaacact 300gaatcagctt gctgacgtta
gaggt
32570325DNAArtificialEngineered miRNA precursor 70ggcagagccg tgcccgtctc
atcccctgcc cgtgcaagca gctaggtagg acgatttgag 60cgtggtgtta ggccgaaccg
ctgaaggaag attgctccac tgttgactgc atttatactc 120caccacaact ggcaaatgta
ttgcttatat tcagcaatat aatgttcgcc agttgtttgg 180gagtataaat atagtcgata
gtggaagaac ggtaacatat gtggtttgca gcaggtgagc 240aggatgggtg tggatgattg
aatatctctg ttcagtgttt tcatcatctg actgaacact 300gaatcagctt gctgacgtta
gaggt
32571408DNAArtificialEngineered miRNA precursor 71ggcagagccg tgcccgtctc
atcccctgcc cgtgcaagca gctaggtagg acgatttgag 60cgtggtgtta ggccgaaccg
ctgaaggaag attgctccac tgttgactgc attgagagat 120ggaacagatg cccaaatgta
ttgcttatat tcagcaatat aatgttcggg catctgggac 180atctctcaat atagtcgata
gtggaagaac ggtaacatat gtggtttgca gcaggtgagc 240aggatgggtg tggatgattg
aatatctctg ttcagtgttt tcatcatctg actgaacact 300gaatcagctt gctgacgtta
gaggtttcag tttacctaat ttatggtctg tacccatgaa 360aagtgggaaa aggctgaaga
attcgatttc tttctttctt tcaatgtt
40872408DNAArtificialEngineered miRNA precursor 72ggcagagccg tgcccgtctc
atcccctgcc cgtgcaagca gctaggtagg acgatttgag 60cgtggtgtta ggccgaaccg
ctgaaggaag attgctccac tgttgactgc attagaagat 120gagaaccctg tgcaaatgta
ttgcttatat tcagcaatat aatgttcgca cagggtgagc 180atcttctaat atagtcgata
gtggaagaac ggtaacatat gtggtttgca gcaggtgagc 240aggatgggtg tggatgattg
aatatctctg ttcagtgttt tcatcatctg actgaacact 300gaatcagctt gctgacgtta
gaggtttcag tttacctaat ttatggtctg tacccatgaa 360aagtgggaaa aggctgaaga
attcgatttc tttctttctt tcaatgtt 4087321DNAArtificialmiRNA
recognition site 73tagatgttga agaggtacca a
217421DNAArtificialmiRNA recognition site 74ttcatagcca
tcacccactt c
217521DNAArtificialmiRNA recognition site 75tgacaagcaa caaagaggtc a
217621DNAArtificialmiRNA
recognition site 76ttatactcca ccacaactgg c
217721DNAArtificialmiRNA recognition site 77tgagagatgg
aacagatgcc c
217821DNAArtificialmiRNA recognition site 78tagaagatga gaaccctgtg c
2179315PRTArabidopsis thaliana
79Met Val Gly Trp Ala Ile Ala Leu His Gly Gly Ala Gly Asp Ile Pro1
5 10 15Ile Asp Leu Pro Asp Glu
Arg Arg Ile Pro Arg Glu Ser Ala Leu Arg 20 25
30His Cys Leu Asp Leu Gly Ile Ser Ala Leu Lys Ser Gly
Lys Pro Pro 35 40 45Leu Asp Val
Ala Glu Leu Val Val Arg Glu Leu Glu Asn His Pro Asp 50
55 60Phe Asn Ala Gly Lys Gly Ser Val Leu Thr Ala Gln
Gly Thr Val Glu65 70 75
80Met Glu Ala Ser Ile Met Asp Gly Lys Thr Lys Arg Cys Gly Ala Val
85 90 95Ser Gly Leu Thr Thr Val
Val Asn Pro Ile Ser Leu Ala Arg Leu Val 100
105 110Met Glu Lys Thr Pro His Ile Tyr Leu Ala Phe Asp
Ala Ala Glu Ala 115 120 125Phe Ala
Arg Ala His Gly Val Glu Thr Val Tyr Ser Ser His Phe Ile 130
135 140Thr Pro Glu Asn Ile Ala Arg Leu Lys Gln Ala
Lys Glu Phe Asn Arg145 150 155
160Val Gln Leu Asp Tyr Thr Val Pro Ser Pro Lys Val Pro Asp Asn Cys
165 170 175Gly Asp Ser Gln
Ile Gly Thr Val Gly Cys Val Ala Val Asp Ser Ala 180
185 190Gly Asn Leu Ala Ser Ala Thr Ser Thr Gly Gly
Tyr Val Asn Lys Met 195 200 205Val
Gly Arg Ile Gly Asp Thr Pro Val Ile Gly Ala Gly Thr Tyr Ala 210
215 220Asn His Leu Cys Ala Ile Ser Ala Thr Gly
Lys Gly Glu Asp Ile Ile225 230 235
240Arg Gly Thr Val Ala Arg Asp Val Ala Ala Leu Met Glu Tyr Lys
Gly 245 250 255Leu Ser Leu
Thr Glu Ala Ala Ala Tyr Val Val Asp Gln Ser Val Pro 260
265 270Arg Gly Ser Cys Gly Leu Val Ala Val Ser
Ala Asn Gly Glu Val Thr 275 280
285Met Pro Phe Asn Thr Thr Gly Met Phe Arg Ala Cys Ala Ser Glu Asp 290
295 300Gly Tyr Ser Glu Ile Ala Ile Trp
Pro Asn Asn305 310 31580726PRTArabidopsis
thaliana 80Met Pro Ser His Pro Asn Phe Ile Phe Arg Trp Ile Gly Leu Phe
Ser1 5 10 15Asp Lys Phe
Arg Arg Gln Thr Thr Gly Ile Asp Glu Asn Ser Asn Leu 20
25 30Gln Ile Asn Gly Gly Asp Ser Ser Ser Ser
Gly Ser Asp Glu Thr Pro 35 40
45Val Leu Ser Ser Ile Glu Cys Tyr Ala Cys Thr Gln Val Gly Val Pro 50
55 60Ala Phe His Ser Thr Ser Cys Asp Gln
Ala His Ala Pro Glu Trp Arg65 70 75
80Ala Ser Ala Gly Ser Ser Leu Val Pro Ile Gln Glu Gly Ser
Val Pro 85 90 95Asn Pro
Ala Arg Thr Arg Phe Arg Arg Leu Lys Gly Pro Phe Gly Glu 100
105 110Val Leu Asp Pro Arg Ser Lys Arg Val
Gln Arg Trp Asn Arg Ala Leu 115 120
125Leu Leu Ala Arg Gly Met Ala Leu Ala Val Asp Pro Leu Phe Phe Tyr
130 135 140Ala Leu Ser Ile Gly Arg Thr
Thr Gly Pro Ala Cys Leu Tyr Met Asp145 150
155 160Gly Ala Phe Ala Ala Val Val Thr Val Leu Arg Thr
Cys Leu Asp Ala 165 170
175Val His Leu Trp His Val Trp Leu Gln Phe Arg Leu Ala Tyr Val Ser
180 185 190Arg Glu Ser Leu Val Val
Gly Cys Gly Lys Leu Val Trp Asp Pro Arg 195 200
205Ala Ile Ala Ser His Tyr Ala Arg Ser Leu Thr Gly Phe Trp
Phe Asp 210 215 220Val Ile Val Ile Leu
Pro Val Pro Gln Ala Val Phe Trp Leu Val Val225 230
235 240Pro Lys Leu Ile Arg Glu Glu Lys Val Lys
Leu Ile Met Thr Ile Leu 245 250
255Leu Leu Ile Phe Leu Phe Gln Phe Leu Pro Lys Ile Tyr His Cys Ile
260 265 270Cys Leu Met Arg Arg
Met Gln Lys Val Thr Gly Tyr Ile Phe Gly Thr 275
280 285Ile Trp Trp Gly Phe Ala Leu Asn Leu Ile Ala Tyr
Phe Ile Ala Ser 290 295 300His Val Ala
Gly Gly Cys Trp Tyr Val Leu Ala Ile Gln Arg Val Ala305
310 315 320Ser Cys Ile Arg Gln Gln Cys
Met Arg Thr Gly Asn Cys Asn Leu Ser 325
330 335Leu Ala Cys Lys Glu Glu Val Cys Tyr Gln Phe Val
Ser Pro Thr Ser 340 345 350Thr
Val Gly Tyr Pro Cys Leu Ser Gly Asn Leu Thr Ser Val Val Asn 355
360 365Lys Pro Met Cys Leu Asp Ser Asn Gly
Pro Phe Arg Tyr Gly Ile Tyr 370 375
380Arg Trp Ala Leu Pro Val Ile Ser Ser Asn Ser Leu Ala Val Lys Ile385
390 395 400Leu Tyr Pro Ile
Phe Trp Gly Leu Met Thr Leu Ser Thr Phe Ala Asn 405
410 415Asp Leu Glu Pro Thr Ser Asn Trp Leu Glu
Val Ile Phe Ser Ile Val 420 425
430Met Val Leu Ser Gly Leu Leu Leu Phe Thr Leu Leu Ile Gly Asn Ile
435 440 445Gln Val Phe Leu His Ala Val
Met Ala Lys Lys Arg Lys Met Gln Ile 450 455
460Arg Cys Arg Asp Met Glu Trp Trp Met Lys Arg Arg Gln Leu Pro
Ser465 470 475 480Arg Leu
Arg Gln Arg Val Arg Arg Phe Glu Arg Gln Arg Trp Asn Ala
485 490 495Leu Gly Gly Glu Asp Glu Leu
Glu Leu Ile His Asp Leu Pro Pro Gly 500 505
510Leu Arg Arg Asp Ile Lys Arg Tyr Leu Cys Phe Asp Leu Ile
Asn Lys 515 520 525Val Pro Leu Phe
Arg Gly Met Asp Asp Leu Ile Leu Asp Asn Ile Cys 530
535 540Asp Arg Ala Lys Pro Arg Val Phe Ser Lys Asp Glu
Lys Ile Ile Arg545 550 555
560Glu Gly Asp Pro Val Gln Arg Met Ile Phe Ile Met Arg Gly Arg Val
565 570 575Lys Arg Ile Gln Ser
Leu Ser Lys Gly Val Leu Ala Thr Ser Thr Leu 580
585 590Glu Pro Gly Gly Tyr Leu Gly Asp Glu Leu Leu Ser
Trp Cys Leu Arg 595 600 605Arg Pro
Phe Leu Asp Arg Leu Pro Pro Ser Ser Ala Thr Phe Val Cys 610
615 620Leu Glu Asn Ile Glu Ala Phe Ser Leu Gly Ser
Glu Asp Leu Arg Tyr625 630 635
640Ile Thr Asp His Phe Arg Tyr Lys Phe Ala Asn Glu Arg Leu Lys Arg
645 650 655Thr Ala Arg Tyr
Tyr Ser Ser Asn Trp Arg Thr Trp Ala Ala Val Asn 660
665 670Ile Gln Met Ala Trp Arg Arg Arg Arg Lys Arg
Thr Arg Gly Glu Asn 675 680 685Ile
Gly Gly Ser Met Ser Pro Val Ser Glu Asn Ser Ile Glu Gly Asn 690
695 700Ser Glu Arg Arg Leu Leu Gln Tyr Ala Ala
Met Phe Met Ser Ile Arg705 710 715
720Pro His Asp His Leu Glu 72581365PRTOryza
sativa Japonica Group 81Ala Gly Ser Asp Glu Val Asn Arg Asn Glu Cys Lys
Thr Val Val Pro1 5 10
15Leu His Thr Trp Val Leu Ile Ser Asn Phe Lys Leu Ser Tyr Asn Ile
20 25 30Leu Arg Arg Ala Asp Gly Thr
Phe Glu Arg Asp Leu Gly Glu Tyr Leu 35 40
45Asp Arg Arg Val Pro Ala Asn Ala Arg Pro Leu Glu Gly Val Ser
Ser 50 55 60Phe Asp His Ile Ile Asp
Gln Ser Val Gly Leu Glu Val Arg Ile Tyr65 70
75 80Arg Ala Ala Ala Glu Gly Asp Ala Glu Glu Gly
Ala Ala Ala Val Thr 85 90
95Arg Pro Ile Leu Glu Phe Leu Thr Asp Ala Pro Ala Ala Glu Pro Phe
100 105 110Pro Val Ile Ile Phe Phe
His Gly Gly Ser Phe Val His Ser Ser Ala 115 120
125Ser Ser Thr Ile Tyr Asp Ser Leu Cys Arg Arg Phe Val Lys
Leu Ser 130 135 140Lys Gly Val Val Val
Ser Val Asn Tyr Arg Arg Ala Pro Glu His Arg145 150
155 160Tyr Pro Cys Ala Tyr Asp Asp Gly Trp Thr
Ala Leu Lys Trp Val Met 165 170
175Ser Gln Pro Phe Met Arg Ser Gly Gly Asp Ala Gln Ala Arg Val Phe
180 185 190Leu Ser Gly Asp Ser
Ser Gly Gly Asn Ile Ala His His Val Ala Val 195
200 205Arg Ala Ala Asp Glu Gly Val Lys Val Cys Gly Asn
Ile Leu Leu Asn 210 215 220Ala Met Phe
Gly Gly Thr Glu Arg Thr Glu Ser Glu Arg Arg Leu Asp225
230 235 240Gly Lys Tyr Phe Val Thr Leu
Gln Asp Arg Asp Trp Tyr Trp Lys Ala 245
250 255Tyr Leu Pro Glu Asp Ala Asp Arg Asp His Pro Ala
Cys Asn Pro Phe 260 265 270Gly
Pro Asn Gly Arg Arg Leu Gly Gly Leu Pro Phe Ala Lys Ser Leu 275
280 285Ile Ile Val Ser Gly Leu Asp Leu Thr
Cys Asp Arg Gln Leu Ala Tyr 290 295
300Ala Asp Ala Leu Arg Glu Asp Gly His His Val Lys Val Val Gln Cys305
310 315 320Glu Asn Ala Thr
Val Gly Phe Tyr Leu Leu Pro Asn Thr Val His Tyr 325
330 335His Glu Val Met Glu Glu Ile Ser Asp Phe
Leu Asn Ala Asn Leu Tyr 340 345
350Tyr Gly Ser His His His His His His His His His His 355
360 36582390PRTZea mays 82Met Gln Ser Ala Ala
Ala Ile Gly Leu Leu Arg Pro Cys Ala Ala Arg1 5
10 15Pro Leu Ala Ala Tyr Thr Ser Pro Arg Arg Gly
Ala Gly Ala Cys Ser 20 25
30Gly Gly Thr Gln Pro Ile Ile Thr Pro Arg Gly Ile Arg Leu Ser Ala
35 40 45Arg Pro Gly Leu Val Pro Ala Ser
Pro Leu Glu Glu Lys Glu Asn Arg 50 55
60Arg Cys Arg Ala Ser Met His Thr Ala Ala Ser Ala Gly Glu Glu Ala65
70 75 80Gly Gly Gly Leu Ala
Lys Thr Leu Gln Leu Gly Ala Leu Phe Gly Leu 85
90 95Trp Tyr Leu Phe Asn Ile Tyr Phe Asn Ile Tyr
Asn Lys Gln Val Leu 100 105
110Lys Val Leu Pro Tyr Pro Ile Asn Ile Thr Thr Val Gln Phe Ala Val
115 120 125Gly Ser Ala Ile Ala Leu Phe
Met Trp Ile Thr Gly Ile His Lys Arg 130 135
140Pro Lys Ile Ser Gly Ala Gln Leu Phe Ala Ile Leu Pro Leu Ala
Ile145 150 155 160Val His
Thr Met Gly Asn Leu Phe Thr Asn Met Ser Leu Gly Lys Val
165 170 175Ala Val Ser Phe Thr His Thr
Ile Lys Ala Met Glu Pro Phe Phe Ser 180 185
190Val Leu Leu Ser Ala Ile Phe Leu Gly Glu Leu Pro Thr Pro
Trp Val 195 200 205Val Leu Ser Leu
Leu Pro Ile Val Gly Gly Val Ala Leu Ala Ser Leu 210
215 220Thr Glu Ala Ser Phe Asn Trp Ala Gly Phe Trp Ser
Ala Met Ala Ser225 230 235
240Asn Val Thr Phe Gln Ser Arg Asn Val Leu Ser Lys Lys Leu Met Val
245 250 255Lys Lys Glu Glu Ser
Leu Asp Asn Ile Asn Leu Phe Ser Ile Ile Thr 260
265 270Val Met Ser Phe Phe Leu Leu Ala Pro Val Thr Leu
Leu Thr Glu Gly 275 280 285Val Lys
Val Ser Pro Ala Val Leu Gln Ser Ala Gly Leu Asn Leu Lys 290
295 300Gln Val Tyr Thr Arg Ser Leu Ile Ala Ala Cys
Cys Phe His Ala Tyr305 310 315
320Gln Gln Val Ser Tyr Met Ile Leu Ala Arg Val Ser Pro Val Thr His
325 330 335Ser Val Gly Asn
Cys Val Lys Arg Val Val Val Ile Val Thr Ser Val 340
345 350Leu Phe Phe Arg Thr Pro Val Ser Pro Ile Asn
Ser Leu Gly Thr Gly 355 360 365Ile
Ala Leu Ala Gly Val Phe Leu Tyr Ser Gln Leu Lys Arg Leu Lys 370
375 380Pro Lys Pro Lys Thr Ala385
39083504PRTArabidopsis thaliana 83Met Val Ser Leu Leu Ser Phe Phe Leu
Leu Leu Leu Val Pro Ile Phe1 5 10
15Phe Leu Leu Ile Phe Thr Lys Lys Ile Lys Glu Ser Lys Gln Asn
Leu 20 25 30Pro Pro Gly Pro
Ala Lys Leu Pro Ile Ile Gly Asn Leu His Gln Leu 35
40 45Gln Gly Leu Leu His Lys Cys Leu His Asp Leu Ser
Lys Lys His Gly 50 55 60Pro Val Met
His Leu Arg Leu Gly Phe Ala Pro Met Val Val Ile Ser65 70
75 80Ser Ser Glu Ala Ala Glu Glu Ala
Leu Lys Thr His Asp Leu Glu Cys 85 90
95Cys Ser Arg Pro Ile Thr Met Ala Ser Arg Val Phe Ser Arg
Asn Gly 100 105 110Lys Asp Ile
Gly Phe Gly Val Tyr Gly Asp Glu Trp Arg Glu Leu Arg 115
120 125Lys Leu Ser Val Arg Glu Phe Phe Ser Val Lys
Lys Val Gln Ser Phe 130 135 140Lys Tyr
Ile Arg Glu Glu Glu Asn Asp Leu Met Ile Lys Lys Leu Lys145
150 155 160Glu Leu Ala Ser Lys Gln Ser
Pro Val Asp Leu Ser Lys Ile Leu Phe 165
170 175Gly Leu Thr Ala Ser Ile Ile Phe Arg Thr Ala Phe
Gly Gln Ser Phe 180 185 190Phe
Asp Asn Lys His Val Asp Gln Glu Ser Ile Lys Glu Leu Met Phe 195
200 205Glu Ser Leu Ser Asn Met Thr Phe Arg
Phe Ser Asp Phe Phe Pro Thr 210 215
220Ala Gly Leu Lys Trp Phe Ile Gly Phe Val Ser Gly Gln His Lys Arg225
230 235 240Leu Tyr Asn Val
Phe Asn Arg Val Asp Thr Phe Phe Asn His Ile Val 245
250 255Asp Asp His His Ser Lys Lys Ala Thr Gln
Asp Arg Pro Asp Met Val 260 265
270Asp Ala Ile Leu Asp Met Ile Asp Asn Glu Gln Gln Tyr Ala Ser Phe
275 280 285Lys Leu Thr Val Asp His Leu
Lys Gly Val Leu Ser Asn Ile Tyr His 290 295
300Ala Gly Ile Asp Thr Ser Ala Ile Thr Leu Ile Trp Ala Met Ala
Glu305 310 315 320Leu Val
Arg Asn Pro Arg Val Met Lys Lys Ala Gln Asp Glu Ile Arg
325 330 335Thr Cys Ile Gly Ile Lys Gln
Glu Gly Arg Ile Met Glu Glu Asp Leu 340 345
350Asp Lys Leu Gln Tyr Leu Lys Leu Val Val Lys Glu Thr Leu
Arg Leu 355 360 365His Pro Ala Ala
Pro Leu Leu Leu Pro Arg Glu Thr Met Ala Asp Ile 370
375 380Lys Ile Gln Gly Tyr Asp Ile Pro Gln Lys Arg Ala
Leu Leu Val Asn385 390 395
400Ala Trp Ser Ile Gly Arg Asp Pro Glu Ser Trp Lys Asn Pro Glu Glu
405 410 415Phe Asn Pro Glu Arg
Phe Ile Asp Cys Pro Val Asp Tyr Lys Gly His 420
425 430Ser Cys Glu Leu Leu Pro Phe Gly Ser Gly Arg Arg
Ile Cys Pro Gly 435 440 445Ile Ala
Met Ala Ile Ala Thr Ile Glu Leu Gly Leu Leu Asn Leu Leu 450
455 460Tyr Phe Phe Asp Trp Asn Met Pro Glu Lys Lys
Lys Asp Met Asp Met465 470 475
480Glu Glu Ala Gly Asp Leu Thr Val Asp Lys Lys Val Pro Leu Glu Leu
485 490 495Leu Pro Val Ile
Arg Ile Ser Leu 50084387PRTPseudomonas syringae pv. tomato
str. DC3000 84Met Thr Val Leu Lys Met Thr Asp Leu Asp Leu Gln Gly Lys Arg
Val1 5 10 15Leu Ile Arg
Glu Asp Leu Asn Val Pro Ile Lys Asp Gly Val Val Ser 20
25 30Ser Asp Ala Arg Ile Leu Ala Ser Leu Pro
Thr Ile Arg Leu Ala Leu 35 40
45Glu Lys Gly Ala Ala Val Met Val Cys Ser His Leu Gly Arg Pro Thr 50
55 60Glu Gly Glu Phe Ser Ala Glu Asn Ser
Leu Lys Pro Val Ala Glu Tyr65 70 75
80Leu Ser Lys Ala Leu Gly Arg Asp Val Pro Leu Val Ala Asp
Tyr Leu 85 90 95Asp Gly
Val Asp Val Lys Ala Gly Asp Ile Val Leu Phe Glu Asn Val 100
105 110Arg Phe Asn Lys Gly Glu Lys Lys Asn
Ala Asp Glu Leu Ala Gln Lys 115 120
125Tyr Ala Ala Leu Cys Asp Val Phe Val Met Asp Ala Phe Gly Thr Ala
130 135 140His Arg Ala Glu Gly Ser Thr
His Gly Val Ala Lys Tyr Ala Lys Val145 150
155 160Ala Ala Ala Gly Pro Leu Leu Ala Ala Glu Leu Glu
Ala Leu Gly Lys 165 170
175Ala Leu Gly Ala Pro Ala Gln Pro Met Ala Ala Ile Val Ala Gly Ser
180 185 190Lys Val Ser Thr Lys Leu
Asp Val Leu Asn Ser Leu Ser Ala Ile Cys 195 200
205Asp Gln Leu Ile Val Gly Gly Gly Ile Ala Asn Thr Phe Leu
Ala Ala 210 215 220Ala Gly His Lys Val
Gly Lys Ser Leu Tyr Glu Pro Asp Leu Leu Asp225 230
235 240Thr Ala Arg Ala Ile Ala Ala Lys Val Ser
Val Pro Leu Pro Thr Asp 245 250
255Val Val Val Ala Lys Glu Phe Ala Glu Ser Ala Thr Ala Thr Val Lys
260 265 270Leu Ile Ala Asp Val
Ala Asp Asp Asp Met Ile Leu Asp Ile Gly Pro 275
280 285Gln Thr Ala Ala His Phe Ala Glu Leu Leu Lys Ser
Ser Gly Thr Ile 290 295 300Leu Trp Asn
Gly Pro Val Gly Val Phe Glu Phe Asp Gln Phe Gly Glu305
310 315 320Gly Thr Lys Thr Leu Ala Lys
Ala Ile Ala Glu Ser Lys Ala Phe Ser 325
330 335Ile Ala Gly Gly Gly Asp Thr Leu Ala Ala Ile Asp
Lys Tyr Gly Val 340 345 350Ala
Asp Gln Ile Ser Tyr Ile Ser Thr Gly Gly Gly Ala Phe Leu Glu 355
360 365Phe Val Glu Gly Lys Val Leu Pro Ala
Val Glu Met Leu Glu Gln Arg 370 375
380Ala Arg Ala38585387PRTPseudomonas syringae pv. phaseolicola 1448A
85Met Thr Val Leu Lys Met Thr Asp Leu Asp Leu Gln Gly Lys Arg Val1
5 10 15Leu Ile Arg Glu Asp Leu
Asn Val Pro Val Lys Asp Gly Val Val Ser 20 25
30Ser Asp Ala Arg Ile Leu Ala Ser Leu Pro Thr Ile Arg
Leu Ala Leu 35 40 45Glu Lys Gly
Ala Ala Val Met Val Cys Ser His Leu Gly Arg Pro Thr 50
55 60Glu Gly Glu Phe Ser Ala Glu Asn Ser Leu Lys Pro
Val Ala Asp Tyr65 70 75
80Leu Ser Lys Ala Leu Gly Arg Asp Val Pro Leu Val Ala Asp Tyr Leu
85 90 95Asp Gly Val Asp Val Lys
Ala Gly Asp Val Val Leu Phe Glu Asn Val 100
105 110Arg Phe Asn Lys Gly Glu Lys Lys Asn Ala Asp Glu
Leu Ala Gln Lys 115 120 125Tyr Ala
Ala Leu Cys Asp Val Phe Val Met Asp Ala Phe Gly Thr Ala 130
135 140His Arg Ala Glu Gly Ser Thr His Gly Val Ala
Lys Phe Ala Lys Val145 150 155
160Ala Ala Ala Gly Pro Leu Leu Ala Ala Glu Leu Glu Ala Leu Gly Lys
165 170 175Ala Leu Gly Ala
Pro Ala Gln Pro Met Thr Ala Ile Val Ala Gly Ser 180
185 190Lys Val Ser Thr Lys Leu Asp Val Leu Asn Ser
Leu Ser Gly Ile Cys 195 200 205Asn
Gln Leu Ile Val Gly Gly Gly Ile Ala Asn Thr Phe Leu Ala Ala 210
215 220Ala Gly His Lys Val Gly Lys Ser Leu Tyr
Glu Pro Asp Leu Leu Asp225 230 235
240Thr Ala Arg Ala Ile Ala Ala Lys Val Ser Val Pro Leu Pro Thr
Asp 245 250 255Val Val Val
Ala Lys Glu Phe Ala Glu Ser Ala Thr Ala Thr Val Lys 260
265 270Leu Ile Ala Asp Val Ala Asp Asp Asp Met
Ile Leu Asp Ile Gly Pro 275 280
285Gln Thr Ala Ala His Phe Ala Glu Leu Leu Lys Ser Ser Gly Thr Ile 290
295 300Leu Trp Asn Gly Pro Val Gly Val
Phe Glu Phe Asp Gln Phe Gly Glu305 310
315 320Gly Thr Lys Thr Leu Ala Lys Ala Ile Gly Glu Ser
Gln Ala Phe Ser 325 330
335Ile Ala Gly Gly Gly Asp Thr Leu Ala Ala Ile Asp Lys Tyr Gly Val
340 345 350Ala Glu Gln Ile Ser Tyr
Ile Ser Thr Gly Gly Gly Ala Phe Leu Glu 355 360
365Phe Val Glu Gly Lys Val Leu Pro Ala Val Glu Val Leu Glu
Gln Arg 370 375 380Ala Lys
Ala38586387PRTPseudomonas syringae pv. syringae B728a 86Met Thr Val Leu
Lys Met Thr Asp Leu Asp Leu Gln Gly Lys Arg Val1 5
10 15Leu Ile Arg Glu Asp Leu Asn Val Pro Val
Lys Asp Gly Val Val Ser 20 25
30Ser Asp Ala Arg Ile Leu Ala Ser Leu Pro Thr Ile Arg Leu Ala Leu
35 40 45Glu Lys Gly Ala Ala Val Met Val
Cys Ser His Leu Gly Arg Pro Thr 50 55
60Glu Gly Glu Phe Ser Ala Glu Asn Ser Leu Lys Pro Val Ala Asp Tyr65
70 75 80Leu Ser Lys Ala Leu
Gly Arg Asp Val Pro Leu Val Ala Asp Tyr Leu 85
90 95Asp Gly Val Asp Val Lys Ala Gly Glu Val Val
Leu Phe Glu Asn Val 100 105
110Arg Phe Asn Lys Gly Glu Lys Lys Asn Ala Asp Glu Leu Ala Gln Gln
115 120 125Tyr Ala Ala Leu Cys Asp Val
Phe Val Met Asp Ala Phe Gly Thr Ala 130 135
140His Arg Ala Glu Gly Ser Thr His Gly Val Ala Lys Phe Ala Lys
Val145 150 155 160Ala Ala
Ala Gly Pro Leu Leu Ala Ala Glu Leu Glu Ala Leu Gly Lys
165 170 175Ala Leu Gly Ala Pro Ala Gln
Pro Met Thr Ala Ile Val Ala Gly Ser 180 185
190Lys Val Ser Thr Lys Leu Asp Val Leu Asn Ser Leu Ser Gly
Ile Cys 195 200 205Asn Gln Leu Ile
Val Gly Gly Gly Ile Ala Asn Thr Phe Leu Ala Ala 210
215 220Ala Gly His Lys Val Gly Lys Ser Leu Tyr Glu Pro
Asp Leu Leu Asp225 230 235
240Thr Ala Arg Ala Ile Ala Ala Lys Val Ser Val Pro Leu Pro Thr Asp
245 250 255Val Val Val Ala Lys
Glu Phe Ala Glu Ser Ala Ala Ala Thr Val Lys 260
265 270Leu Ile Ala Asp Val Ala Asp Asp Asp Met Ile Leu
Asp Ile Gly Pro 275 280 285Gln Thr
Ala Ala His Phe Ala Glu Leu Leu Lys Ser Ser Gly Thr Ile 290
295 300Leu Trp Asn Gly Pro Val Gly Val Phe Glu Phe
Asp Gln Phe Gly Glu305 310 315
320Gly Thr Lys Thr Leu Ala Lys Ala Ile Ala Glu Ser Gln Ala Phe Ser
325 330 335Ile Ala Gly Gly
Gly Asp Thr Leu Ala Ala Ile Asp Lys Tyr Gly Val 340
345 350Ala Gln Gln Ile Ser Tyr Ile Ser Thr Gly Gly
Gly Ala Phe Leu Glu 355 360 365Phe
Val Glu Gly Lys Val Leu Pro Ala Val Glu Val Leu Glu Gln Arg 370
375 380Ala Lys Ala38587393PRTSorghum bicolor
87Met Ser Leu Ile Arg Gly Met Gly Asn Ile Ala Lys Arg Trp Lys Glu1
5 10 15Leu Asn Gly Leu Asn Tyr
Trp Lys Gly Leu Val Asp Pro Leu Asp Leu 20 25
30Asp Leu Arg Arg Asn Ile Ile Asn Tyr Gly Glu Leu Ser
Gln Ala Ala 35 40 45Tyr Thr Gly
Leu Asn Arg Glu Arg Arg Ser Arg Tyr Ala Gly Ser Cys 50
55 60Leu Phe Asn Arg Arg Asp Phe Leu Ser Arg Val Asp
Val Ser Asn Pro65 70 75
80Asn Leu Tyr Glu Ile Thr Lys Phe Ile Tyr Ala Met Cys Thr Val Ser
85 90 95Leu Pro Asp Gly Phe Met
Val Lys Ser Leu Ser Lys Ala Ala Trp Ser 100
105 110Arg Gln Ser Asn Trp Met Gly Phe Val Ala Val Ala
Thr Asp Glu Gly 115 120 125Lys Glu
Val Leu Gly Arg Arg Asp Val Val Val Ala Trp Arg Gly Thr 130
135 140Ile Arg Met Val Glu Trp Met Asp Asp Leu Asp
Ile Ser Leu Val Pro145 150 155
160Ala Ser Glu Ile Val Leu Pro Gly Ser Ala Thr Asn Pro Cys Val His
165 170 175Gly Gly Trp Leu
Ser Val Tyr Thr Ser Ala Asp Pro Gly Ser Gln Tyr 180
185 190Asn Lys Glu Ser Ala Arg His Gln Val Leu Asn
Glu Val Lys Arg Ile 195 200 205Gln
Asp Leu Tyr Lys Thr Glu Glu Thr Ser Ile Ser Ile Thr Gly His 210
215 220Ser Leu Gly Ala Ala Leu Ala Thr Ile Asn
Ala Ile Asp Ile Val Ser225 230 235
240Asn Gly Tyr Asn Arg Ser Cys Pro Val Ser Ala Phe Val Phe Gly
Ser 245 250 255Pro Arg Val
Gly Asn Pro Asp Phe Gln Glu Ala Phe Asp Ser Ala Ala 260
265 270Asp Leu Arg Leu Leu Arg Val Arg Asn Ser
Pro Asp Val Val Pro Lys 275 280
285Trp Pro Lys Leu Gly Tyr Ser Asp Val Gly Thr Glu Leu Arg Ile Asp 290
295 300Thr Gly Glu Ser Pro Tyr Leu Lys
Ser Pro Gly Asn Pro Leu Thr Trp305 310
315 320His Asp Met Glu Cys Tyr Met His Gly Val Ala Gly
Ala Gln Gly Ser 325 330
335Ser Gly Gly Phe Glu Leu Ala Val Asp Arg Asp Ile Ala Leu Val Asn
340 345 350Lys His Glu Asp Ala Leu
Lys Asn Glu Phe Ala Val Pro Ser Ser Trp 355 360
365Trp Val Val Gln Asn Lys Asp Met Val Lys Gly Lys Asp Gly
Arg Trp 370 375 380His Leu Ala Asp His
Glu Asp Asp Asp385 39088512PRTArabidopsis thaliana 88Met
Ala Thr Leu Leu Ala Thr Pro Ile Phe Ser Pro Leu Ala Ser Ser1
5 10 15Pro Ala Arg Asn Arg Leu Ser
Cys Ser Asn Ile Arg Phe Gly Ser Lys 20 25
30Asn Gly Lys Ile Leu Asn Ser Asp Gly Ala Gln Lys Leu Asn
Leu Ser 35 40 45Lys Phe Arg Lys
Pro Asp Gly Gln Arg Phe Leu Gln Met Gly Ser Ser 50 55
60Lys Glu Met Asn Phe Glu Arg Lys Leu Ser Val Gln Ala
Met Asp Gly65 70 75
80Ala Gly Thr Gly Asn Thr Ser Thr Ile Ser Arg Asn Val Ile Ala Ile
85 90 95Ser His Leu Leu Val Ser
Leu Gly Ile Ile Leu Ala Ala Asp Tyr Phe 100
105 110Leu Lys Gln Ala Phe Val Ala Ala Ser Ile Lys Phe
Pro Ser Ala Leu 115 120 125Phe Gly
Met Phe Cys Ile Phe Ser Val Leu Met Ile Phe Asp Ser Val 130
135 140Val Pro Ala Ala Ala Asn Gly Leu Met Asn Phe
Phe Glu Pro Ala Phe145 150 155
160Leu Phe Ile Gln Arg Trp Leu Pro Leu Phe Tyr Val Pro Ser Leu Val
165 170 175Val Leu Pro Leu
Ser Val Arg Asp Ile Pro Ala Ala Ser Gly Val Lys 180
185 190Ile Cys Tyr Ile Val Ala Gly Gly Trp Leu Ala
Ser Leu Cys Val Ala 195 200 205Gly
Tyr Thr Ala Ile Ala Val Arg Lys Met Val Lys Thr Glu Met Thr 210
215 220Glu Ala Glu Pro Met Ala Lys Pro Ser Pro
Phe Ser Thr Leu Glu Leu225 230 235
240Trp Ser Trp Ser Gly Ile Phe Val Val Ser Phe Val Gly Ala Leu
Phe 245 250 255Tyr Pro Asn
Ser Leu Gly Thr Ser Ala Arg Thr Ser Leu Pro Phe Leu 260
265 270Leu Ser Ser Thr Val Leu Gly Tyr Ile Val
Gly Ser Gly Leu Pro Ser 275 280
285Ser Ile Lys Lys Val Phe His Pro Ile Ile Cys Cys Ala Leu Ser Ala 290
295 300Val Leu Ala Ala Leu Ala Phe Gly
Tyr Ala Ser Gly Ser Gly Leu Asp305 310
315 320Pro Val Leu Gly Asn Tyr Leu Thr Lys Val Ala Ser
Asp Pro Gly Ala 325 330
335Gly Asp Ile Leu Met Gly Phe Leu Gly Ser Val Ile Leu Ser Phe Ala
340 345 350Phe Ser Met Phe Lys Gln
Arg Lys Leu Val Lys Arg His Ala Ala Glu 355 360
365Ile Phe Thr Ser Val Ile Val Ser Thr Val Phe Ser Leu Tyr
Ser Thr 370 375 380Ala Leu Val Gly Arg
Leu Val Gly Leu Glu Pro Ser Leu Thr Val Ser385 390
395 400Ile Leu Pro Arg Cys Ile Thr Val Ala Leu
Ala Leu Ser Ile Val Ser 405 410
415Leu Phe Glu Gly Thr Asn Ser Ser Leu Thr Ala Ala Val Val Val Val
420 425 430Thr Gly Leu Ile Gly
Ala Asn Phe Val Gln Val Val Leu Asp Lys Leu 435
440 445Arg Leu Arg Asp Pro Ile Ala Arg Gly Ile Ala Thr
Ala Ser Ser Ala 450 455 460His Gly Leu
Gly Thr Ala Ala Leu Ser Ala Lys Glu Pro Glu Ala Leu465
470 475 480Pro Phe Cys Ala Ile Ala Tyr
Ala Leu Thr Gly Ile Phe Gly Ser Leu 485
490 495Leu Cys Ser Val Pro Ala Val Arg Gln Ser Leu Leu
Ala Val Val Gly 500 505
51089311PRTZea mays 89Met Ala Arg Asn Glu Glu Lys Ala Gln Ser Met Leu Asn
Arg Phe Ile1 5 10 15Thr
Met Lys Gln Glu Glu Lys Arg Lys Pro Arg Glu Arg Arg Pro Tyr 20
25 30Leu Ala Ser Glu Cys Arg Asp Leu
Ala Asp Ala Glu Arg Trp Arg Ser 35 40
45Glu Ile Leu Arg Glu Ile Gly Ala Lys Val Ala Glu Ile Gln Asn Glu
50 55 60Gly Leu Gly Glu His Arg Leu Arg
Asp Leu Asn Asp Glu Ile Asn Lys65 70 75
80Leu Leu Arg Glu Arg Gly His Trp Glu Arg Arg Ile Val
Glu Leu Gly 85 90 95Gly
Arg Asp Tyr Ser Arg Ser Ser Asn Ala Pro Leu Met Thr Asp Leu
100 105 110Asp Gly Asn Ile Val Ala Val
Pro Asn Pro Ser Gly Arg Gly Pro Gly 115 120
125Tyr Arg Tyr Phe Gly Ala Ala Arg Lys Leu Pro Gly Val Arg Glu
Leu 130 135 140Phe Asp Lys Pro Pro Glu
Met Arg Lys Arg Arg Thr Arg Tyr Glu Ile145 150
155 160His Lys Arg Ile Asn Ala Gly Tyr Tyr Gly Tyr
Tyr Asp Asp Glu Asp 165 170
175Gly Val Leu Glu Arg Leu Glu Gly Pro Ala Glu Lys Arg Met Arg Glu
180 185 190Glu Ile Val Ser Glu Trp
His Arg Val Glu Arg Val Arg Arg Glu Ala 195 200
205Met Lys Gly Val Met Ser Gly Glu Val Ala Ala Ala Gly Gly
Arg Ser 210 215 220Gly Glu Ala Ala Arg
Glu Val Leu Phe Glu Gly Val Glu Glu Glu Val225 230
235 240Glu Glu Glu Arg Lys Arg Glu Glu Glu Lys
Arg Glu Arg Glu Lys Gly 245 250
255Glu Glu Val Gly Arg Glu Phe Val Ala His Val Pro Leu Pro Asp Glu
260 265 270Lys Glu Ile Glu Arg
Met Val Leu Glu Arg Lys Lys Lys Glu Leu Leu 275
280 285Ser Lys Tyr Ala Ser Asp Ser Leu Leu Val Glu Gln
Glu Glu Ala Lys 290 295 300Glu Met Leu
Asn Val Arg Arg305 31090309PRTSorghum bicolor 90Met Ala
Arg Asn Glu Glu Lys Ala Gln Ser Met Leu Asn Arg Phe Ile1 5
10 15Thr Met Lys Gln Glu Glu Lys Arg
Lys Pro Arg Glu Arg Arg Pro Tyr 20 25
30Leu Ala Ser Glu Cys Arg Asp Leu Ala Asp Ala Glu Arg Trp Arg
Ser 35 40 45Glu Ile Leu Arg Glu
Ile Gly Ala Lys Val Ala Glu Ile Gln Asn Glu 50 55
60Gly Leu Gly Glu His Arg Leu Arg Asp Leu Asn Asp Glu Ile
Asn Lys65 70 75 80Leu
Leu Arg Glu Arg Gly His Trp Glu Arg Arg Ile Val Glu Leu Gly
85 90 95Gly Arg Asp Tyr Ser Arg Ser
Ser Asn Ala Pro Leu Met Thr Asp Leu 100 105
110Asp Gly Asn Ile Val Ala Val Pro Asn Pro Ser Gly Arg Gly
Pro Gly 115 120 125Tyr Arg Tyr Phe
Gly Ala Ala Arg Lys Leu Pro Gly Val Arg Glu Leu 130
135 140Phe Asp Lys Pro Pro Glu Met Arg Lys Arg Arg Thr
Arg Tyr Glu Ile145 150 155
160His Lys Arg Ile Asn Ala Gly Tyr Tyr Gly Tyr Tyr Asp Asp Glu Asp
165 170 175Gly Val Leu Glu Arg
Leu Glu Ala Pro Ala Glu Lys Arg Met Arg Glu 180
185 190Glu Ile Val Ser Glu Trp His Arg Val Glu Arg Val
Arg Arg Glu Ala 195 200 205Met Lys
Gly Val Val Ser Gly Glu Val Ala Ala Ala Gly Gly Arg Ser 210
215 220Gly Glu Ala Ala Arg Glu Val Leu Phe Glu Gly
Val Glu Glu Glu Val225 230 235
240Glu Glu Glu Arg Lys Arg Glu Glu Glu Lys Arg Glu Arg Glu Lys Gly
245 250 255Glu Glu Ala Glu
Phe Val Ala His Val Pro Leu Pro Asp Glu Lys Glu 260
265 270Ile Glu Arg Met Val Leu Glu Arg Lys Lys Lys
Glu Leu Leu Ser Lys 275 280 285Tyr
Ala Ser Asp Ser Leu Leu Val Glu Gln Glu Glu Ala Lys Glu Met 290
295 300Leu Asn Val Arg Arg30591453PRTArabidopsis
thalianamisc_feature(327)..(327)Xaa can be any naturally occurring amino
acid 91Met Ala Lys Val Tyr Trp Pro Tyr Phe Asp Pro Glu Tyr Glu Asn Leu1
5 10 15Ser Ser Arg Ile Asn
Pro Pro Ser Val Ser Ile Asp Asn Thr Ser Cys 20
25 30Lys Glu Cys Thr Leu Val Lys Val Asp Ser Met Asn
Lys Pro Gly Ile 35 40 45Leu Leu
Glu Val Val Gln Val Leu Thr Asp Leu Asp Leu Thr Ile Thr 50
55 60Lys Ala Tyr Ile Ser Ser Asp Gly Gly Trp Phe
Met Asp Val Phe His65 70 75
80Val Thr Asp Gln Gln Gly Asn Lys Val Thr Asp Ser Lys Thr Ile Asp
85 90 95Tyr Ile Glu Lys Val
Leu Gly Pro Lys Gly His Ala Ser Ala Ser Gln 100
105 110Asn Thr Trp Pro Gly Lys Arg Val Gly Val His Ser
Leu Gly Asp His 115 120 125Thr Ser
Ile Glu Ile Ile Ala Arg Asp Arg Pro Gly Leu Leu Ser Glu 130
135 140Val Ser Ala Val Leu Ala Asp Leu Asn Ile Asn
Val Val Ala Ala Glu145 150 155
160Ala Trp Thr His Asn Arg Arg Ile Ala Cys Val Leu Tyr Val Asn Asp
165 170 175Asn Ala Thr Ser
Arg Ala Val Asp Asp Pro Glu Arg Leu Ser Ser Met 180
185 190Glu Glu Gln Leu Asn Asn Val Leu Arg Gly Cys
Glu Glu Gln Asp Glu 195 200 205Lys
Phe Ala Arg Thr Ser Leu Ser Ile Gly Ser Thr His Val Asp Arg 210
215 220Arg Leu His Gln Met Phe Phe Ala Asp Arg
Asp Tyr Glu Ala Val Thr225 230 235
240Lys Leu Asp Asp Ser Ala Ser Cys Gly Phe Glu Pro Lys Ile Thr
Val 245 250 255Glu His Cys
Glu Glu Lys Gly Tyr Ser Val Ile Asn Val Ser Cys Glu 260
265 270Asp Arg Pro Lys Leu Met Phe Asp Ile Val
Cys Thr Leu Thr Asp Met 275 280
285Gln Tyr Ile Val Phe His Ala Thr Ile Ser Ser Ser Gly Ser His Ala 290
295 300Ser Gln Glu Tyr Phe Ile Arg His
Lys Asp Gly Cys Thr Leu Asp Thr305 310
315 320Glu Gly Glu Lys Glu Arg Xaa Val Lys Cys Leu Glu
Ala Ala Ile His 325 330
335Arg Arg Val Ser Glu Gly Trp Ser Leu Glu Leu Cys Ala Lys Asp Arg
340 345 350Val Gly Leu Leu Ser Glu
Val Thr Arg Ile Leu Arg Glu His Gly Leu 355 360
365Ser Val Ser Arg Ala Gly Val Thr Thr Val Gly Glu Gln Ala
Val Asn 370 375 380Val Phe Tyr Val Lys
Asp Ala Ser Gly Asn Pro Val Asp Val Lys Thr385 390
395 400Ile Glu Ala Leu Arg Gly Glu Ile Gly His
Ser Met Met Ile Asp Phe 405 410
415Lys Asn Lys Val Pro Ser Arg Lys Trp Lys Glu Glu Gly Gln Ala Gly
420 425 430Thr Gly Gly Gly Trp
Ala Lys Thr Ser Phe Phe Phe Gly Asn Leu Leu 435
440 445Glu Lys Leu Leu Pro 45092143PRTArabidopsis
thaliana 92Met Gln Glu Leu Gly Leu Gln Arg Phe Ser Asn Asp Val Val Arg
Leu1 5 10 15Asp Leu Thr
Pro Pro Ser Gln Thr Ser Ser Thr Ser Leu Ser Ile Asp 20
25 30Glu Glu Glu Ser Thr Glu Ala Lys Ile Arg
Arg Leu Ile Ser Glu His 35 40
45Pro Val Ile Ile Phe Ser Arg Ser Ser Cys Cys Met Cys His Val Met 50
55 60Lys Arg Leu Leu Ala Thr Ile Gly Val
Ile Pro Thr Val Ile Glu Leu65 70 75
80Asp Asp His Glu Val Ser Ser Leu Pro Thr Ala Leu Gln Asp
Glu Tyr 85 90 95Ser Gly
Gly Val Ser Val Val Gly Pro Pro Pro Ala Val Phe Ile Gly 100
105 110Arg Glu Cys Val Gly Gly Leu Glu Ser
Leu Val Ala Leu His Leu Ser 115 120
125Gly Gln Leu Val Pro Lys Leu Val Gln Val Gly Ala Leu Trp Val 130
135 14093942PRTZea mays 93Met Glu Gly Asp
Asp Phe Thr Pro Glu Gly Gly Lys Leu Pro Glu Phe1 5
10 15Lys Leu Asp Ala Arg Gln Ala Gln Gly Phe
Ile Ser Phe Phe Lys Lys 20 25
30Leu Pro Gln Asp Pro Arg Ala Val Arg Leu Phe Asp Arg Arg Asp Tyr
35 40 45Tyr Thr Ala His Gly Glu Asn Ala
Thr Phe Ile Ala Arg Thr Tyr Tyr 50 55
60His Thr Met Ser Ala Leu Arg Gln Leu Gly Ser Ser Ser Asp Gly Ile65
70 75 80Leu Ser Ala Ser Val
Ser Lys Ala Met Phe Glu Thr Ile Ala Arg Asn 85
90 95Ile Leu Leu Glu Arg Thr Asp Cys Thr Leu Glu
Leu Tyr Glu Gly Ser 100 105
110Gly Ser Asn Trp Arg Leu Thr Lys Ser Gly Thr Pro Gly Asn Ile Gly
115 120 125Ser Phe Glu Asp Ile Leu Phe
Ala Asn Asn Asp Met Glu Asp Ser Pro 130 135
140Val Ile Val Ala Leu Phe Pro Ala Cys Arg Glu Ser Gln Leu Tyr
Val145 150 155 160Gly Leu
Ser Phe Leu Asp Met Thr Asn Arg Lys Leu Gly Leu Ala Glu
165 170 175Phe Pro Glu Asp Ser Arg Phe
Thr Asn Val Glu Ser Ala Leu Val Ala 180 185
190Leu Gly Cys Lys Glu Cys Leu Leu Pro Ala Asp Cys Glu Lys
Ser Ile 195 200 205Asp Leu Asn Pro
Leu Gln Asp Val Ile Ser Asn Cys Asn Val Leu Leu 210
215 220Thr Glu Lys Lys Lys Ala Asp Phe Lys Ser Arg Asp
Leu Ala Gln Asp225 230 235
240Leu Gly Arg Ile Ile Arg Gly Ser Val Glu Pro Val Arg Asp Leu Leu
245 250 255Ser Gln Phe Asp Tyr
Ala Leu Gly Pro Leu Gly Ala Leu Leu Ser Tyr 260
265 270Ala Glu Leu Leu Ala Asp Asp Thr Asn Tyr Gly Asn
Tyr Thr Ile Glu 275 280 285Lys Tyr
Asn Leu Asn Cys Tyr Met Arg Leu Asp Ser Ala Ala Val Arg 290
295 300Ala Leu Asn Ile Ala Glu Gly Lys Thr Asp Val
Asn Lys Asn Phe Ser305 310 315
320Leu Phe Gly Leu Met Asn Arg Thr Cys Thr Val Gly Met Gly Lys Arg
325 330 335Leu Leu Asn Arg
Trp Leu Lys Gln Pro Leu Leu Asp Val Asn Glu Ile 340
345 350Asn Asn Arg Leu Asp Met Val Gln Ala Phe Val
Glu Asp Pro Glu Leu 355 360 365Arg
Gln Gly Leu Arg Gln Gln Leu Lys Arg Ile Ser Asp Ile Asp Arg 370
375 380Leu Thr His Ser Leu Arg Lys Lys Ser Ala
Asn Leu Gln Pro Val Val385 390 395
400Lys Leu Tyr Gln Ser Cys Ser Arg Ile Pro Tyr Ile Lys Gly Ile
Leu 405 410 415Gln Gln Tyr
Asn Gly Gln Phe Ser Thr Leu Ile Arg Ser Lys Phe Leu 420
425 430Glu Pro Leu Glu Glu Trp Met Ala Lys Asn
Arg Phe Gly Arg Phe Ser 435 440
445Ser Leu Val Glu Thr Ala Ile Asp Leu Ala Gln Leu Glu Asn Gly Glu 450
455 460Tyr Arg Ile Ser Pro Leu Tyr Ser
Ser Asp Leu Gly Val Leu Lys Asp465 470
475 480Glu Leu Ser Val Val Glu Asn His Ile Asn Asn Leu
His Val Asp Thr 485 490
495Ala Ser Asp Leu Asp Leu Ser Val Asp Lys Gln Leu Lys Leu Glu Lys
500 505 510Gly Ser Leu Gly His Val
Phe Arg Met Ser Lys Lys Glu Glu Gln Lys 515 520
525Val Arg Lys Lys Leu Thr Gly Ser Tyr Leu Ile Ile Glu Thr
Arg Lys 530 535 540Asp Gly Val Lys Phe
Thr Asn Ser Lys Leu Lys Asn Leu Ser Asp Gln545 550
555 560Tyr Gln Ala Leu Phe Gly Glu Tyr Thr Ser
Cys Gln Lys Lys Val Val 565 570
575Gly Asp Val Val Arg Val Ser Gly Thr Phe Ser Glu Val Phe Glu Asn
580 585 590Phe Ala Ala Val Leu
Ser Glu Leu Asp Val Leu Gln Ser Phe Ala Asp 595
600 605Leu Ala Thr Ser Cys Pro Val Pro Tyr Val Arg Pro
Asp Ile Thr Ala 610 615 620Ser Asp Glu
Gly Asp Ile Val Leu Leu Gly Ser Arg His Pro Cys Leu625
630 635 640Glu Ala Gln Asp Gly Val Asn
Phe Ile Pro Asn Asp Cys Thr Leu Val 645
650 655Arg Gly Lys Ser Trp Phe Gln Ile Ile Thr Gly Pro
Asn Met Gly Gly 660 665 670Lys
Ser Thr Phe Ile Arg Gln Val Gly Val Asn Val Leu Met Ala Gln 675
680 685Val Gly Ser Phe Val Pro Cys Asp Gln
Ala Ser Ile Ser Val Arg Asp 690 695
700Cys Ile Phe Ala Arg Val Gly Ala Gly Asp Cys Gln Leu His Gly Val705
710 715 720Ser Thr Phe Met
Gln Glu Met Leu Glu Thr Ala Ser Ile Leu Lys Gly 725
730 735Ala Ser Asp Lys Ser Leu Ile Ile Ile Asp
Glu Leu Gly Arg Gly Thr 740 745
750Ser Thr Tyr Asp Gly Phe Gly Leu Ala Trp Ala Ile Cys Glu His Leu
755 760 765Met Glu Val Thr Arg Ala Pro
Thr Leu Phe Ala Thr His Phe His Glu 770 775
780Leu Thr Ala Leu Ala His Arg Asn Asp Asp Glu His Gln His Ile
Ser785 790 795 800Asp Ile
Gly Val Ala Asn Tyr His Val Gly Ala His Ile Asp Pro Leu
805 810 815Ser Arg Lys Leu Thr Met Leu
Tyr Lys Val Glu Pro Gly Ala Cys Asp 820 825
830Gln Ser Phe Gly Ile His Val Ala Glu Phe Ala Asn Phe Pro
Glu Ala 835 840 845Val Val Ala Leu
Ala Lys Ser Lys Ala Ala Glu Leu Glu Asp Phe Ser 850
855 860Thr Thr Pro Thr Phe Ser Asp Asp Leu Lys Asp Glu
Val Gly Ser Lys865 870 875
880Arg Lys Arg Val Phe Ser Pro Asp Asp Ile Thr Arg Gly Ala Ala Arg
885 890 895Ala Arg Leu Phe Leu
Glu Glu Phe Ala Ala Leu Pro Met Asp Glu Met 900
905 910Asp Gly Ser Lys Ile Leu Glu Met Ala Thr Lys Met
Lys Ala Asp Leu 915 920 925Gln Lys
Asp Ala Ala Asp Asn Pro Trp Leu Gln Gln Phe Phe 930
935 94094942PRTSorghum bicolor 94Met Glu Gly Asp Asp Phe
Thr Pro Glu Gly Gly Lys Leu Pro Glu Phe1 5
10 15Lys Leu Asp Ala Arg Gln Ala Gln Gly Phe Ile Ser
Phe Phe Lys Arg 20 25 30Leu
Pro Gln Asp Pro Arg Ala Val Arg Leu Phe Asp Arg Arg Asp Tyr 35
40 45Tyr Thr Ala His Gly Glu Asn Ala Thr
Phe Ile Ala Arg Thr Tyr Tyr 50 55
60His Thr Met Ser Ala Leu Arg Gln Leu Gly Ser Ser Ser Asp Gly Ile65
70 75 80Ser Ser Val Ser Val
Ser Lys Ala Met Phe Glu Thr Ile Ala Arg Asn 85
90 95Ile Leu Leu Glu Arg Thr Asp Cys Thr Leu Glu
Leu Tyr Glu Gly Ser 100 105
110Gly Ser Asn Trp Arg Leu Thr Lys Ser Gly Thr Pro Gly Asn Ile Gly
115 120 125Ser Phe Glu Asp Leu Leu Phe
Ala Asn Asn Asp Met Gln Asp Ser Pro 130 135
140Val Ile Val Ala Leu Phe Pro Val Cys Arg Glu Ser Gln Leu Tyr
Val145 150 155 160Gly Leu
Ser Phe Leu Asp Met Thr Asn Arg Lys Leu Gly Leu Ala Glu
165 170 175Phe Pro Glu Asp Ser Arg Phe
Thr Asn Val Glu Ser Ala Leu Val Ala 180 185
190Leu Gly Cys Lys Glu Cys Leu Leu Ser Glu Asp Cys Glu Lys
Ser Ile 195 200 205Asp Leu Asn Pro
Leu Arg Asp Ala Ile Ser Asn Cys Asn Val Leu Leu 210
215 220Thr Val Lys Lys Lys Ala Asp Phe Lys Ser Arg Asp
Leu Ala Gln Asp225 230 235
240Leu Gly Arg Ile Ile Arg Gly Ser Val Glu Pro Val Arg Asp Leu Leu
245 250 255Ser Gln Phe Asp Tyr
Ala Leu Gly Pro Leu Gly Ala Leu Leu Ser Tyr 260
265 270Ala Glu Leu Leu Ala Asp Asp Thr Asn Tyr Gly Asn
Tyr Thr Ile Glu 275 280 285Lys Tyr
Asn Leu Asn Cys Tyr Met Arg Leu Asp Ser Ala Ala Val Arg 290
295 300Ala Leu Asn Ile Ser Glu Arg Lys Thr Asp Val
Asn Lys Asn Phe Ser305 310 315
320Leu Phe Gly Leu Met Asn Arg Thr Cys Thr Val Gly Met Gly Lys Arg
325 330 335Leu Leu Asn Arg
Trp Leu Lys Gln Pro Leu Leu Asp Val Asn Glu Ile 340
345 350Asn Asn Arg Leu Asp Met Val Gln Ala Phe Val
Glu Asp Pro Glu Leu 355 360 365Arg
Gln Gly Leu Arg Gln Gln Leu Lys Arg Ile Ser Asp Ile Asp Arg 370
375 380Leu Thr His Ala Leu Arg Lys Lys Ser Ala
Thr Leu Gln Pro Val Val385 390 395
400Lys Leu Tyr Gln Ser Cys Cys Arg Ile Ser Tyr Ile Lys Gly Ile
Leu 405 410 415Glu Gln Tyr
Asn Gly Gln Phe Ser Thr Leu Ile Arg Ser Lys Phe Leu 420
425 430Glu Pro Leu Glu Glu Trp Met Ala Glu Asp
Arg Phe Gly Arg Phe Ser 435 440
445Ser Leu Val Glu Thr Thr Ile Asp Leu Gly Gln Leu Glu Asn Gly Glu 450
455 460Tyr Arg Ile Ser Pro Leu Tyr Ser
Ser Asp Leu Gly Val Leu Lys Asp465 470
475 480Glu Leu Ser Val Val Glu Asn His Ile Asn Asn Leu
His Val Asp Thr 485 490
495Ala Ser Asp Leu Asp Leu Ser Val Asp Lys Gln Leu Lys Leu Glu Lys
500 505 510Gly Pro Leu Gly His Val
Phe Arg Met Ser Lys Lys Glu Glu Gln Lys 515 520
525Val Arg Lys Lys Leu Thr Gly Ser Tyr Leu Ile Ile Glu Thr
Arg Lys 530 535 540Asp Gly Val Lys Phe
Thr Ser Ser Lys Leu Lys Lys Leu Ser Asp Gln545 550
555 560Tyr Gln Ala Leu Phe Ala Glu Tyr Thr Ser
Cys Gln Lys Lys Val Val 565 570
575Gly Asp Val Val Arg Val Ser Gly Ser Tyr Ser Glu Val Phe Glu Asn
580 585 590Phe Ala Ala Val Leu
Ser Glu Leu Asp Val Leu Gln Ser Phe Ala Asp 595
600 605Leu Ala Thr Ser Cys Pro Val Pro Tyr Val Arg Pro
Asp Ile Thr Val 610 615 620Ser Asp Glu
Gly Asp Ile Val Leu Leu Gly Ser Arg His Pro Cys Leu625
630 635 640Glu Ala Gln Asp Gly Val Asn
Phe Ile Pro Asn Asp Cys Thr Leu Val 645
650 655Arg Gly Lys Ser Trp Phe Gln Ile Ile Thr Gly Pro
Asn Met Gly Gly 660 665 670Lys
Ser Thr Phe Ile Arg Gln Val Gly Val Asn Val Leu Met Ala Gln 675
680 685Val Gly Ser Phe Val Pro Cys Asp Gln
Ala Ser Val Ser Val Arg Asp 690 695
700Cys Ile Phe Ala Arg Val Gly Ala Gly Asp Cys Gln Leu His Gly Val705
710 715 720Ser Thr Phe Met
Gln Glu Met Leu Glu Thr Ala Ser Ile Leu Lys Gly 725
730 735Ala Ser Asp Lys Ser Leu Ile Ile Ile Asp
Glu Leu Gly Arg Gly Thr 740 745
750Ser Thr Tyr Asp Gly Phe Gly Leu Ala Trp Ala Ile Cys Glu His Leu
755 760 765Met Glu Val Thr Arg Ala Pro
Thr Leu Phe Ala Thr His Phe His Glu 770 775
780Leu Thr Ala Leu Ala His Lys Asn Asp Asp Glu His Gln Arg Val
Ser785 790 795 800Asn Ile
Gly Ile Ala Asn Tyr His Val Gly Ala His Ile Asp Pro Ser
805 810 815Ser Arg Lys Leu Thr Met Leu
Tyr Lys Val Glu Pro Gly Ala Cys Asp 820 825
830Gln Ser Phe Gly Ile His Val Ala Glu Phe Ala Asn Phe Pro
Glu Ala 835 840 845Val Val Ala Leu
Ala Lys Ser Lys Ala Ala Glu Leu Glu Asp Phe Ser 850
855 860Thr Thr Pro Thr Phe Ser Asp Asp Ser Lys Asp Glu
Val Gly Ser Lys865 870 875
880Arg Lys Arg Val Phe Ser Pro Asp Asp Val Thr Arg Gly Ala Ala Arg
885 890 895Ala Arg Leu Phe Leu
Glu Asp Phe Ala Ala Leu Pro Val Asp Glu Met 900
905 910Asp Arg Ser Lys Ile Val Glu Met Val Thr Lys Met
Lys Ser Asp Leu 915 920 925Gln Lys
Asp Ala Ala Asp Asn Pro Trp Leu Gln Gln Phe Phe 930
935 94095274PRTGlycine max 95Met Arg Ala Lys Leu Phe Val
Phe Pro Ile Arg Gly Arg Asn Trp Cys1 5 10
15Phe Ser Arg Thr Ile Asp His Ser Leu Ser Ala Ser His
Ala Ser Ser 20 25 30Gln Ser
Pro Ser Thr Leu Lys Asp Leu Trp Thr Asn Ile Asn Val Gly 35
40 45Asp Lys Pro Leu Asn Thr Lys Thr Glu Leu
Phe Val Asp Tyr Ile Ala 50 55 60Asn
Lys Met Asn Asn Ala Trp Ile Gly Leu Glu Lys Ala Pro Glu Gly65
70 75 80Ser Phe Lys Asn Lys Ile
His Gly Leu Gly Leu Arg Leu Leu Ser Arg 85
90 95Val Lys Pro Ser Glu Ile Phe Leu Lys Ser Ile Ser
Lys Glu Ile Thr 100 105 110Ser
Val Glu Ile Ile Tyr Pro Ser Ser Leu Asn Ala Gln Leu Val Arg 115
120 125Arg Arg Leu Arg His Ile Ala Val Arg
Gly Ala Val Ile His Arg Asn 130 135
140Tyr Leu Tyr Gly Leu Val Ser Leu Ile Pro Leu Thr Ser Ala Leu Ser145
150 155 160Ile Leu Pro Leu
Pro Asn Val Pro Phe Phe Trp Val Leu Phe Arg Thr 165
170 175Tyr Ser His Trp Arg Ala Leu Gln Gly Ser
Glu Arg Leu Phe Gln Leu 180 185
190Val Ser Asp Asn Ser Lys Thr Ser Asn Thr Cys Thr Tyr Glu Lys Lys
195 200 205Thr Glu His Lys Glu Ser Lys
Ser Gln Arg His Ser Ser Asn Glu Pro 210 215
220Cys Trp Val Leu Arg Pro Ser Lys Glu Leu Glu Asn Leu Val His
Leu225 230 235 240Glu Asp
Gly Gln Glu Ser Phe Ser Gln His Ala Ile Ile Asn Ile Cys
245 250 255Lys Ile Tyr Asp Leu Asn Pro
Val Asp Val Ile Lys Tyr Glu Lys Ser 260 265
270Val Phe96390PRTZea mays 96Met Gln Ser Ala Ala Ala Ile Gly
Leu Leu Arg Pro Cys Ala Ala Arg1 5 10
15Pro Leu Ala Ala Tyr Thr Ser Pro Arg Arg Gly Ala Gly Ala
Cys Ser 20 25 30Gly Gly Thr
Gln Pro Ile Ile Thr Pro Arg Gly Ile Arg Leu Ser Ala 35
40 45Arg Pro Gly Leu Val Pro Ala Ser Pro Leu Glu
Glu Lys Glu Asn Arg 50 55 60Arg Cys
Arg Ala Ser Met His Ala Ala Ala Ser Ala Gly Glu Glu Ala65
70 75 80Gly Gly Gly Leu Ala Lys Thr
Leu Gln Leu Gly Ala Leu Phe Gly Leu 85 90
95Trp Tyr Leu Phe Asn Ile Tyr Phe Asn Ile Tyr Asn Lys
Gln Val Leu 100 105 110Lys Val
Leu Pro Tyr Pro Ile Asn Ile Thr Thr Val Gln Phe Ala Val 115
120 125Gly Ser Ala Ile Ala Leu Phe Met Trp Ile
Thr Gly Ile His Lys Arg 130 135 140Pro
Lys Ile Ser Gly Ala Gln Leu Phe Ala Ile Leu Pro Leu Ala Ile145
150 155 160Val His Thr Met Gly Asn
Leu Phe Thr Asn Met Ser Leu Gly Lys Val 165
170 175Ala Val Ser Phe Thr His Thr Ile Lys Ala Met Glu
Pro Phe Phe Ser 180 185 190Val
Leu Leu Ser Ala Ile Phe Leu Gly Glu Leu Pro Thr Pro Trp Val 195
200 205Val Leu Ser Leu Leu Pro Ile Val Gly
Gly Val Ala Leu Ala Ser Leu 210 215
220Thr Glu Ala Ser Phe Asn Trp Ala Gly Phe Trp Ser Ala Met Ala Ser225
230 235 240Asn Val Thr Phe
Gln Ser Arg Asn Val Leu Ser Lys Lys Leu Met Val 245
250 255Lys Lys Glu Glu Ser Leu Asp Asn Ile Asn
Leu Phe Ser Ile Ile Thr 260 265
270Val Met Ser Phe Phe Leu Leu Ala Pro Val Thr Leu Leu Thr Glu Gly
275 280 285Val Lys Val Ser Pro Ala Val
Leu Gln Ser Ala Gly Leu Asn Leu Lys 290 295
300Gln Val Tyr Thr Arg Ser Leu Ile Ala Ala Phe Cys Phe His Ala
Tyr305 310 315 320Gln Gln
Val Ser Tyr Met Ile Leu Ala Arg Val Ser Pro Val Thr His
325 330 335Ser Val Gly Asn Cys Val Lys
Arg Val Val Val Ile Val Thr Ser Val 340 345
350Leu Phe Phe Arg Thr Pro Val Ser Pro Ile Asn Ser Leu Gly
Thr Gly 355 360 365Ile Ala Leu Ala
Gly Val Phe Leu Tyr Ser Gln Leu Lys Arg Leu Lys 370
375 380Pro Lys Pro Lys Thr Ala385
390972092DNAArtificialCold inducible promoter 97cagatccacg ctcgctcggg
tgtcgggtca gatcgatcca gttggcgcac gtaataatcc 60ttttccccag aaggagtcga
acccctcctc cccgtccaat ccaatcaaag cgaccaatcg 120actggctgtc ctacacacac
acaaaaccga ccgaggcgac acaccgcagc agtgatcatt 180ctgagcattt gcagaaaaag
gagaacgtcc cgaaatcctg gtggttgtat tgtgtgattg 240ctcactcagt ccgtgcaggg
tcagggtgaa gccaagccaa caacccaacg ctcgctggga 300gtagggtcca ccggatttat
tggcagtaca tcgctgtttg gtcctcctgc ccttcgctta 360ttttttaatt cggcagacgt
gcacagacag ggcaccaccg gaccaaggaa gggcgcacac 420cgtcgtcagt caccaggtgg
gtgtgatcag cagccgcttc tcttgtgctg ctttatagcg 480tatgaaattc cagtgtccct
gttccacctg catgcaattg gtttgactga acaacatgat 540agcaagtgat actatatata
tttttataga ggaacacagt gaaaaaatat ttagtattat 600tacgtgcatg aaattgtatt
cacagttatc cctgatgcaa cgcaattgtt caatatatag 660cagtatatat tatacgaagt
atatatgtat atctaatttt atgagaccgg gagaaggtgt 720attcacagta cagtgcaggg
ccatggccat gcagcccttg gggcctgaaa agggtcgcgt 780gaagtggcca acgctgtgca
attgcaacca aacaaacttt tggtggcggg gtccctgtcc 840ctggccggct ttgcccacag
gccacagcgc atcacaccac cgctttatag cgccacccca 900ccaccctcgt ctctcccccc
gtcgagcaca caacacaccc tcctcgtcct ccaatccaat 960caacctggta gactcgcttc
gcttctcccc ccagctcgga cggagctcct cgcagcagcc 1020gccgatcaac ctgcgctcgg
gctcagcgct ggaaggtgag agctcagtgc ctcgtcccgc 1080ccgccccaaa tctggttctt
gtgctggctc tggctgtgcg ctgcacgaat tctgcatctg 1140gttctttcga gacgcaattc
ccggaccgtg ggctttggtt tcggaggggg ccgagagtaa 1200ggcgttagga ctttctccga
gctgcaaggc cgctcgtcgt tgcggcattt ttcgtttcgc 1260ttgtcctgtg atgagagatg
tgcatttccc tttggcgggc ttaccgttcc ctgctcgtct 1320gtatgtgtgt atgtttgtgt
gacctttccc tcaacgccag gctcttctcc cctcttgctg 1380tttctttcag cagtacagac
gcgcatctgt acagcgcctt tcttcggtcc tgggttatga 1440ttgatccgtt aacagttggt
caccaagtgc tggctgttta atatgtacta taagcttctt 1500ggtgccgctg cctctgccta
tacgacttta tgcgctgcct gcacaagtct cagccatctg 1560tgggaacgtg tgtctctcac
ctacctttca tattgcacta gctggattga atcattctgc 1620tttggagaga tgtccggtca
ttttttttta aatcattttc atctcgcgta ctagtttttg 1680ttttgttttg cgagagagta
attttttttt aatatttact gtctcctgtc ccatttgctg 1740tttctttacc cagaaatttc
caccagattc agtcaaacga aactcctgtg ctcttttttt 1800tctccctttc aaaagggtgt
gtaaccgact accgactcag ataatataag tgcggtcaca 1860tatcacatga tatcatctcg
cctctctccc ttctcctgtg ttttattttc cttttttcta 1920accacagcgt gatgaacttc
tttttttttt gggggggggg gggggggtaa ctacagctta 1980gcgaacatga atgggtagtt
ttacaactaa tgcaacggct ggttcactga acaactgtag 2040gtgttggaag agaatagcct
gaaggttcac agtaaccttc atctgtcgga ag 2092982516DNAArtificialSeed
preferred promoter 98acacttttat tatcgcgtca aatcagtacc tcaatcgata
ttgtagccta gtgttcttat 60taaatgggaa gaattcgagg acacactaat tccttgctaa
cacacactta tgctccattt 120ggatgtcgat attggagggc atggaactga attggtttca
attacaaatc agccatgata 180ttgtaatgag atgtaatttc aattctattc tttggatgtc
actgaattgg agtttggaat 240tgtgtggtcc aattccacct tatatagaag agggatgctc
tgtattggga gagtgagttt 300ctagttatag tctagcttcg ggaaattgag tctctcgttc
caaatctcaa ttccatgtgc 360aaccaaacaa tagaattctg gaaagctgat tccaattcct
aattccgtgc tccaatatct 420acatccaaac gggtgttaca taaatataga aatgacatat
caaccatgca aaaccacatt 480ggcgatgttg aacaaaggcg aacacccaca tactatgtac
cgcacacggc atctctttct 540caaaggtcga accacgtgtg ttccatgcat gcgtggaaca
tgcaaggttg tcacgtatag 600ggaatgatga cacacgagag cgcctacaag gcaacaaaca
ccttacgtac cacgtagagt 660gcattttgct accacctgcc accggatgac atgtatgcat
gcatgcgttg tgtacgcata 720cactgctgtc tgctggtgcc caaagaccat ctagaacagc
atcttttaat tctccatttc 780cctcacgcca ttgctagtgc cttgcacatg ctcgcactcc
ctaacacatc ttcctccctt 840tatttttcgt tgccaattgc tagttgttca aatgccacgt
tttccttaca cagctgtagg 900gcaccgtacc acgtagaatg cattcctcgc caccaacaga
caacacggcc gggcatatgt 960acgtcttacg ccggaccatc accagtatat atgatgctag
ggatcagtgg gcgccctttt 1020tgcctcgtcc tcccggggcg gcattcctat gtcctaactg
aagcaaccca cgcgccgcca 1080tttctgttgc gaatgagtcc atggacatat gtgccaacag
aacccctcgg aaggcaccat 1140ctatctatct atctctcaag caatattata tttggcacct
acgctcaagt acatagacag 1200tgtgcacggc attgtgcagc tggaaagccc gcccgacacg
agggctgcca aatcgacagc 1260tccgcgccct tggaaatcct agtcacttgt tcacaattga
ccaatctacc cttgaagcac 1320acggtggatg gtactgccac atttggctta taggggcata
gaggacaatg aatgcaactg 1380gagcgggaag gagagcttta atttgtaagt actcggtgaa
cacggcacct gatgatgatg 1440atgatggaca gcgaggaatt gttataaaag gcgcccgtcc
ctcccatggc tcaagaacaa 1500gggaatcgaa gccattccct cttcaagagg ggatcatcag
attgggctta ttattcctta 1560ttactccagg taattcttag tttgttgccc ttccaaacct
ttacatctca tataagaatg 1620attattacat gcaagattat gttgacatgc gtcgtcatgg
tatttttttt aggcaaggat 1680cggagttgct ctgaattgac tgaaccagat ctaccgtctt
cggtacgcgc tcactccgcc 1740ctctgccttt gttactgcca cgtttctctg aatgctctct
tgtgtggtga ttgctgagag 1800tggtttagct ggatctagaa ttacactctg aaatcgtgtt
ctgcctgtgc tgattacttg 1860ccgtcctttg tagcagcaaa atatagggac atggtagtac
gaaacgaaga tagaacctac 1920acagcaatac gagaaatgtg taatttggtg cttagcggta
tttatttaag cacatgttgg 1980tgttataggg cacttggatt cagaagtttg ctgttaattt
aggcacaggc ttcatactac 2040atgggtcaat agtataggga ttcatattat aggcgatact
ataataattt gttcgtctgc 2100agagcttatt atttgccaaa attagatatt cctattctgt
ttttgtttgt gtgctgttaa 2160attgttaacg cctgaaggaa taaatataaa tgacgaaatt
ttgatgttta tctctgctcc 2220tttattgtga ccataagtca agatcagatg cacttgtttt
aaatattgtt gtctgaagaa 2280ataagtactg acagtatttt gatgcattga tctgcttgtt
tgttgtaaca aaatttaaaa 2340ataaagagtt tcctttttgt tgctctcctt acctcctgat
ggtatctagt atctaccaac 2400tgacactata ttgcttctct ttacatacgt atcttgctcg
atgccttctc cctagtgttg 2460accagtgtta ctcacatagt ctttgctcat ttcattgtaa
tgcagatacc aagcgg 2516991761DNAArtificialLeaf preferred promoter
99gacatggagg tggaaggcct gacgtagata gagaagatgc tcttagcttt cattgtcttt
60cttttgtagt catctgattt acctctctcg tttatacaac tggtttttta aacactcctt
120aacttttcaa attgtctctt tctttaccct agactagata attttaatgg tgattttgct
180aatgtggcgc catgttagat agaggtaaaa tgaactagtt aaaagctcag agtgataaat
240caggctctca aaaattcata aactgttttt taaatatcca aatattttta catggaaaat
300aataaaattt agtttagtat taaaaaattc agttgaatat agttttgtct tcaaaaatta
360tgaaactgat cttaattatt tttccttaaa accgtgctct atctttgatg tctagtttga
420gacgattata taattttttt tgtgcttaac tacgacgagc tgaagtacgt agaaatacta
480gtggagtcgt gccgcgtgtg cctgtagcca ctcgtacgct acagcccaag cgctagagcc
540caagaggccg gaggtggaag gcgtcgcggc actatagcca ctcgccgcaa gagcccaaga
600gaccggagct ggaaggatga gggtctgggt gttcacgaat tgcctggagg caggaggctc
660gtcgtccgga gccacaggcg tggagacgtc cgggataagg tgagcagccg ctgcgatagg
720ggcgcgtgtg aaccccgtcg cgccccacgg atggtataag aataaaggca ttccgcgtgc
780aggattcacc cgttcgcctc tcaccttttc gctgtactca ctcgccacac acaccccctc
840tccagctccg ttggagctcc ggacagcagc aggcgcgggg cggtcacgta gtaagcagct
900ctcggctccc tctccccttg ctccatttga tagtgcaacc catcgagcta cgggcccacc
960gtcttcggta cgcgctcact ccgccctctg cctttgttac tgccacgttt ctctgaatgc
1020tctcttgtgt ggtgattgct gagagtggtt tagctggatc tagaattaca ctctgaaatc
1080gtgttctgcc tgtgctgatt acttgccgtc ctttgtagca gcaaaatata gggacatggt
1140agtacgaaac gaagatagaa cctacacagc aatacgagaa atgtgtaatt tggtgcttag
1200cggtatttat ttaagcacat gttggtgtta tagggcactt ggattcagaa gtttgctgtt
1260aatttaggca caggcttcat actacatggg tcaatagtat agggattcat attataggcg
1320atactataat aatttgttcg tctgcagagc ttattatttg ccaaaattag atattcctat
1380tctgtttttg tttgtgtgct gttaaattgt taacgcctga aggaataaat ataaatgacg
1440aaattttgat gtttatctct gctcctttat tgtgaccata agtcaagatc agatgcactt
1500gttttaaata ttgttgtctg aagaaataag tactgacagt attttgatgc attgatctgc
1560ttgtttgttg taacaaaatt taaaaataaa gagtttcctt tttgttgctc tccttacctc
1620ctgatggtat ctagtatcta ccaactgaca ctatattgct tctctttaca tacgtatctt
1680gctcgatgcc ttctccctag tgttgaccag tgttactcac atagtctttg ctcatttcat
1740tgtaatgcag ataccaagcg g
17611002924DNAArtificialLeaf preferred promoter 100atgtgctggt gccccataag
gtaggcacct aggtctgtgt ttgaagcatc gacagatttg 60taaacatgtt cctatgaacc
tatttctgat tgataatttg tcaaaactca tcatttgtct 120tcatccttgc ctgcttgcgt
tcacgtgaca aagtacgtgt atgtcttcgg cctttgctgt 180gtatgtttcg cattgcttag
atgtggtgaa agaacatcag aagatgcatt gatggcgtgc 240ttaaaccagt gatgtgctcc
aggtgttcct gcagtctgca gagatattta ctcttgtagt 300cttgttgaca gcacagttgt
atgtgatttc ttggatgtaa tgtaaaccaa atgaaagata 360ggaacagttc gtcctcttcc
gtatacgaag gtcactgtat catttgtcgt ggcacaagat 420gatctgcagg caggactgca
acatggtttc ttggactgtc ctgaatgccc gttcttgttc 480tttagttgag ccagagcagc
agcctggtgt cggtgcctga gacctgacga agcacacggc 540aaacaaacaa gtcgcagcag
ctagcagggg cgttgccatc gccacaagcc cccaagagac 600ccgccgagga aaagaaaaaa
aaactacggc cgccgttgcc aagccgagcg tgcgaaccga 660tccacggatg ggagatcaga
gatcacccac cgcaggcggg cggcagtggc tggcgaggtg 720cgtccacaga acctgctgca
ggtccctgtc cgtcccggcg accccttttc taggcgagca 780actccccatg gcagagctgc
acgcagcagg gcccgtcgtt ggttgcagct ttaacccttt 840ttgttttaac catacaatgc
agagtcgcag aggtgaaaca ggacggaaat tacagaaaag 900atggtggtgt gccagcagcc
ccagcatgaa gaagatcagg acaaaagaaa agcttgtgat 960tggtgacagc aacaggattg
gattggagcc aagctaggca gtgagaggca ggcagcaaga 1020cgcgtcagcc actgaaatcc
agagggcaac ctcggcctca caactcatat ccccttgtgc 1080tgttgcgcgc cgtggttagc
caggtgtgct gcagggggta ccatggcatg catcgataga 1140tctcgaggga tccaaagaca
tggaggtgga aggcctgacg tagatagaga agatgctctt 1200agctttcatt gtctttcttt
tgtagtcatc tgatttacct ctctcgttta tacaactggt 1260tttttaaaca ctccttaact
tttcaaattg tctctttctt taccctagac tagataattt 1320taatggtgat tttgctaatg
tggcgccatg ttagatagag gtaaaatgaa ctagttaaaa 1380gctcagagtg ataaatcagg
ctctcaaaaa ttcataaact gttttttaaa tatccaaata 1440tttttacatg gaaaataata
aaatttagtt tagtattaaa aaattcagtt gaatatagtt 1500ttgtcttcaa aaattatgaa
actgatctta attatttttc cttaaaaccg tgctctatct 1560ttgatgtcta gtttgagacg
attatataat tttttttgtg cttaactacg acgagctgaa 1620gtacgtagaa atactagtgg
agtcgtgccg cgtgtgcctg tagccactcg tacgctacag 1680cccaagcgct agagcccaag
aggccggagg tggaaggcgt cgcggcacta tagccactcg 1740ccgcaagagc ccaagagacc
ggagctggaa ggatgagggt ctgggtgttc acgaattgcc 1800tggaggcagg aggctcgtcg
tccggagcca caggcgtgga gacgtccggg ataaggtgag 1860cagccgctgc gataggggcg
cgtgtgaacc ccgtcgcgcc ccacggatgg tataagaata 1920aaggcattcc gcgtgcagga
ttcacccgtt cgcctctcac cttttcgctg tactcactcg 1980ccacacacac cccctctcca
gctccgttgg agctccggac agcagcaggc gcggggcggt 2040cacgtagtaa gcagctctcg
gctccctctc cccttgctcc atttgatagt gcaacccatc 2100gagctacacc ggtgcggccc
accgtcttcg gtacgcgctc actccgccct ctgcctttgt 2160tactgccacg tttctctgaa
tgctctcttg tgtggtgatt gctgagagtg gtttagctgg 2220atctagaatt acactctgaa
atcgtgttct gcctgtgctg attacttgcc gtcctttgta 2280gcagcaaaat atagggacat
ggtagtacga aacgaagata gaacctacac agcaatacga 2340gaaatgtgta atttggtgct
tagcggtatt tatttaagca catgttggtg ttatagggca 2400cttggattca gaagtttgct
gttaatttag gcacaggctt catactacat gggtcaatag 2460tatagggatt catattatag
gcgatactat aataatttgt tcgtctgcag agcttattat 2520ttgccaaaat tagatattcc
tattctgttt ttgtttgtgt gctgttaaat tgttaacgcc 2580tgaaggaata aatataaatg
acgaaatttt gatgtttatc tctgctcctt tattgtgacc 2640ataagtcaag atcagatgca
cttgttttaa atattgttgt ctgaagaaat aagtactgac 2700agtattttga tgcattgatc
tgcttgtttg ttgtaacaaa atttaaaaat aaagagtttc 2760ctttttgttg ctctccttac
ctcctgatgg tatctagtat ctaccaactg acactatatt 2820gcttctcttt acatacgtat
cttgctcgat gccttctccc tagtgttgac cagtgttact 2880cacatagtct ttgctcattt
cattgtaatg cagataccaa gcgg 2924101408DNAOryza sativa
101ggcagagccg tgcccgtctc atcccctgcc cgtgcaagca gctaggtagg acgatttgag
60cgtggtgtta ggccgaaccg ctgaaggaag attgctccac tgttgactgc attaggattc
120aatccttgct gctaaatgta ttgcttatat tcagcaatat aatgttcagc agcaagaact
180ggatcttaat atagtcgata gtggaagaac ggtaacatat gtggtttgca gcaggtgagc
240aggatgggtg tggatgattg aatatctctg ttcagtgttt tcatcatctg actgaacact
300gaatcagctt gctgacgtta gaggtttcag tttacctaat ttatggtctg tacccatgaa
360aagtgggaaa aggctgaaga attcgatttc tttctttctt tcaatgtt
408102325DNAOryza sativa 102ggcagagccg tgcccgtctc atcccctgcc cgtgcaagca
gctaggtagg acgatttgag 60cgtggtgtta ggccgaaccg ctgaaggaag attgctccac
tgttgactgc attaggattc 120aatccttgct gctaaatgta ttgcttatat tcagcaatat
aatgttcagc agcaagaact 180ggatcttaat atagtcgata gtggaagaac ggtaacatat
gtggtttgca gcaggtgagc 240aggatgggtg tggatgattg aatatctctg ttcagtgttt
tcatcatctg actgaacact 300gaatcagctt gctgacgtta gaggt
325103280DNAOryza sativa 103ggcagagccg tgcccgtctc
atcccctgcc cgtgcaagca gctaggtagg acgatttgag 60cgtggtgtta ggccgaaccg
ctgaaggaag attgctccac tgttgactgc attaggattc 120aatccttgct gctaaatgta
ttgcttatat tcagcaatat aatgttcagc agcaagaact 180ggatcttaat atagtcgata
gtggaagaac ggtaacatat gtggtttgca gcaggtgagc 240aggatgggtg tggatgattg
aatatctctg ttcagtgttt 280104249DNAOryza sativa
104ggcagagccg tgcccgtctc atcccctgcc cgtgcaagca gctaggtagg acgatttgag
60cgtggtgtta ggccgaaccg ctgaaggaag attgctccac tgttgactgc attaggattc
120aatccttgct gctaaatgta ttgcttatat tcagcaatat aatgttcagc agcaagaact
180ggatcttaat atagtcgata gtggaagaac ggtaacatat gtggtttgca gcaggtgagc
240aggatgggt
249
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