Polynucleotides encoding proteins involved in plant metabolism

Abstract

The invention provides isolated pyruvate dehydrogenase kinase nucleic acids and their encoded polypeptides. The present invention provides methods and compositions relating to altering pyruvate dehydrogenase kinase levels in plants. The invention further provides recombinant expression cassettes, host cells, transgenic plants, and antibody compositions.

What is claimed is: 
     
         1 . An isolated nucleic acid encoding a polypeptide selected from the group consisting of SEQ ID NOs:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150, 152, 154, 156, 158, 160, 162, 164, 166, 168, 170, 172, 174, 176, 178, 180, 182, 184, 186, 188, 190, 192, 194, 196, 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 236, 238, 240, 242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264, 266, 268, 270, 272, 274, 276, 278, 280, 282, 284, 286, 288, 290, 292, 294, 296, 298, 300, 302, 304, 306, 308, 310, 312, 314, 316, 318, 320, 322, 324, 326, 328, 330, 332, 334, 336, 338, 340, 342, 344, 346, 348, 350, 352, 354, 356, 358, 360, and 362. 
     
     
         2 . An isolated nucleic acid comprising a member selected from the group consisting of:
 (a) a polynucleotide encoding a polypeptide of at least 50 amino acids that has at least 80% identity based on the Clustal method of alignment when compared to a polypeptide of SEQ ID NO:20;   (b) a polynucleotide encoding a polypeptide of at least 60 amino acids that has at least 95% identity based on the Clustal method of alignment when compared to a polypeptide of SEQ ID NO:6;   (c) a polynucleotide encoding a polypeptide of at least 65 amino acids that has at least 90% identity based on the Clustal method of alignment when compared to a polypeptide of SEQ ID NO:16;   (d) a polynucleotide encoding a polypeptide of at least 95 amino acids that has at least 90% identity based on the Clustal method of alignment when compared to a polypeptide of SEQ ID NO:18;   (e) a polynucleotide encoding a polypeptide of at least 100 amino acids that has at least 85% identity based on the Clustal method of alignment when compared to a polypeptide of SEQ ID NO:12;   (f) a polynucleotide encoding a polypeptide of at least 100 amino acids that has at least 90% identity based on the Clustal method of alignment when compared to a polypeptide of SEQ ID NO:2;   (g) a polynucleotide encoding a polypeptide of at least 100 amino acids that has at least 95% identity based on the Clustal method of alignment when compared to a polypeptide of SEQ ID NO:8;   (h) a polynucleotide encoding a polypeptide of at least 150 amino acids that has at least 90% identity based on the Clustal method of alignment when compared to a polypeptide of SEQ ID NO:22;   (i) a polynucleotide encoding a polypeptide of at least 200 amino acids that has at least 90% identity based on the Clustal method of alignment when compared to a polypeptide of SEQ ID NO:24;   (j) a polynucleotide encoding a polypeptide of at least 200 amino acids that has at least 95% identity based on the Clustal method of alignment when compared to a polypeptide of SEQ ID NO:10;   (k) a polynucleotide encoding a polypeptide of at least 300 amino acids that has at least 90% identity based on the Clustal method of alignment when compared to a polypeptide of SEQ ID NO:4;   (l) a polynucleotide encoding a polypeptide of at least 350 amino acids that has at least 85% identity based on the Clustal method of alignment when compared to a polypeptide of SEQ ID NO:14;   (m) a polynucleotide encoding a polypeptide selected from the group consisting of SEQ ID NOs:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, and 24;   (n) a polynucleotide amplified from a  Zea mays, Oryza sativa, Glycine max , or  Triticum aestivum  nucleic acid library using primers which selectively hybridize, under stringent hybridization conditions, to loci within a polynucleotide selected from the group consisting of SEQ ID NOs:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, and 23;   (o) a polynucleotide which selectively hybridizes, under stringent hybridization conditions and a wash in 2×SSC at 50° C., to a polynucleotide selected from the group consisting of SEQ ID NOs:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, and 23;   (p) a polynucleotide selected from the group consisting of SEQ ID NOs:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, and 23;   (q) a polynucleotide which is complementary to a polynucleotide of (a), (b), (c), (d), (e), (f), (g), (h), (i), (j), (k), (l), (m), (n), (o), or (p); and   (r) a polynucleotide comprising at least 25 contiguous nucleotides from a polynucleotide of (a), (b), (c), (d), (e), (f), (g), (h), (i), (j), (k), (l), (m), (n), (o), (p), or (q).   
     
     
         3 . A recombinant expression cassette, comprising a member of  claim 2  operably linked, in sense or anti-sense orientation, to a promoter. 
     
     
         4 . A host cell comprising the recombinant expression cassette of  claim 3 . 
     
     
         5 . A transgenic plant comprising a recombinant expression cassette of  claim 3 . 
     
     
         6 . The transgenic plant of  claim 5 , wherein said plant is a monocot. 
     
     
         7 . The transgenic plant of  claim 5 , wherein said plant is a dicot. 
     
     
         8 . The transgenic plant of  claim 5 , wherein said plant is selected from the group consisting of: maize, soybean, sunflower, sorghum, canola, wheat, alfalfa, cotton, rice, barley, millet, peanut, and cocoa. 
     
     
         9 . A transgenic seed from the transgenic plant of  claim 5 . 
     
     
         10 . A method of modulating the level of pyruvate dehydrogenase kinase in a plant, comprising:
 (a) introducing into a plant cell a recombinant expression cassette comprising a polynucleotide of  claim 2  operably linked to a promoter;   (b) culturing the plant cell under plant cell growing conditions; and   (c) inducing expression of said polynucleotide for a time sufficient to modulate the level of pyruvate dehydrogenase kinase in said plant.   
     
     
         11 . The method of  claim 10 , wherein the plant is a member of the group consisting of: corn, wheat, rice, and soybean. 
     
     
         12 . An isolated protein comprising a member selected from the group consisting of:
 (a) polypeptide of at least 20 contiguous amino acids from a polypeptide selected from the group consisting of SEQ ID NOs:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, and 24;   (b) a polypeptide selected from the group consisting of SEQ ID NOs:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, and 24;   (c) a polypeptide of at least 50 amino acids that has at least 80% identity based on the Clustal method of alignment when compared to, and having at least one epitope in common with, a polypeptide of SEQ ID NO:20;   (d) a polypeptide of at least 60 amino acids that has at least 95% identity based on the Clustal method of alignment when compared to, and having at least one epitope in common with, a polypeptide of SEQ ID NO:6;   (e) a polypeptide of at least 65 amino acids that has at least 90% identity based on the Clustal method of alignment when compared to, and having at least one epitope in common with, a polypeptide of SEQ ID NO:16;   (f) a polypeptide of at least 95 amino acids that has at least 90% identity based on the Clustal method of alignment when compared to, and having at least one epitope in common with, a polypeptide of SEQ ID NO:18;   (g) a polypeptide of at least 100 amino acids that has at least 85% identity based on the Clustal method of alignment when compared to, and having at least one epitope in common with, a polypeptide of SEQ ID NO:12;   (h) a polypeptide of at least 100 amino acids that has at least 90% identity based on the Clustal method of alignment when compared to, and having at least one epitope in common with, a polypeptide of SEQ ID NO:2;   (i) a polypeptide of at least 100 amino acids that has at least 95% identity based on the Clustal method of alignment when compared to, and having at least one epitope in common with, a polypeptide of SEQ ID NO:8;   (j) a polypeptide of at least 150 amino acids that has at least 90% identity based on the Clustal method of alignment when compared to, and having at least one epitope in common with, a polypeptide of SEQ ID NO:22;   (k) a polypeptide of at least 200 amino acids that has at least 90% identity based on the Clustal method of alignment when compared to, and having at least one epitope in common with, a polypeptide of SEQ ID NO:24;   (l) a polypeptide of at least 200 amino acids that has at least 95% identity based on the Clustal method of alignment when compared to, and having at least one epitope in common with, a polypeptide of SEQ ID NO:10;   (m) a polypeptide of at least 300 amino acids that has at least 90% identity based on the Clustal method of alignment when compared to, and having at least one epitope in common with, a polypeptide of SEQ ID NO:4;   (n) a polypeptide of at least 350 amino acids that has at least 85% identity based on the Clustal method of alignment when compared to, and having at least one epitope in common with, a polypeptide of SEQ ID NO:14; and   (o) at least one polypeptide encoded by a member of  claim 2 .   
     
     
         13 . A data processing system, comprising:
 a set of data representing at least one genetic sequence;   a genetic identification, analysis, or modeling computer program designed to govern the processing of said set of data;   a data processor having an output for storing or displaying data processing results, said data processor containing said data and said program and executing instructions according to said program to process said data or a contiguous subsequence thereof; and   wherein said genetic sequence is: (i) at least 90% sequence identical to a polynucleotide sequence of SEQ ID NOs:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, or 23, or (ii) at least 95% sequence identical to a polypeptide sequence of SEQ ID NOs:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, or 24, and wherein sequence identity is determined by a GAP algorithm under default parameters.