Plant farnesyltransferases

Abstract

This invention relates to an isolated nucleic acid fragment encoding a farnesyltransferase subunit. The invention also relates to the construction of a chimeric gene encoding all or a portion of the farnesyltransferase subunit, in sense or antisense orientation, wherein expression of the chimeric gene results in production of altered levels of the farnesyltransferase subunit in a transformed host cell.

1 . An isolated polynucleotide comprising a nucleotide sequence encoding a first polypeptide of at least 300 amino acids that has at least 80% identity based on the Clustal method of alignment when compared to a polypeptide selected from the group consisting of a corn farnesyltransferase polypeptide of SEQ ID NO:2, a rice farnesyltransferase polypeptide of SEQ ID NO:4, a soybean farnesyltransferase polypeptide of SEQ ID NO:6, a soybean farnesyltransferase polypeptide of SEQ ID NO:8, a wheat farnesyltransferase polypeptide of SEQ ID NO:10, a corn farnesyltransferase polypeptide of SEQ ID NO:12, a rice farnesyltransferase polypeptide of SEQ ID NO:14, a soybean farnesyltransferase polypeptide of SEQ ID NO:16, and a soybean farnesyltransferase polypeptide of SEQ ID NO:18. 
     
     
         2 . An isolated polynucleotide comprising the complement of the polynucleotide of  claim 1 . 
     
     
         3 . The isolated polynucleotide of  claim 1 , wherein the nucleotide sequence comprises a nucleic acid sequence selected from the group consisting of SEQ ID NO:1, 3, 5, 7, 9, 11, 13, 15 and 17 that codes for the polypeptide selected from the group consisting of SEQ ID NO:2, 4, 6, 8, 10, 12, 14, 16 and 18. 
     
     
         4 . The isolated polynucleotide of  claim 1  which is DNA. 
     
     
         5 . The isolated polynucleotide of  claim 1  which is RNA. 
     
     
         6 . A chimeric gene comprising the isolated polynucleotide of  claim 1  or  claim 2  operably linked to suitable regulatory sequences. 
     
     
         7 . An isolated host cell comprising the chimeric gene of  claim 6 . 
     
     
         8 . An isolated host cell comprising an isolated polynucleotide of  claim 1 . 
     
     
         9 . The isolated host cell of  claim 8 , wherein the host cell is yeast. 
     
     
         10 . The isolated host cell of  claim 8 , wherein the host cell is a bacterial cell. 
     
     
         11 . The isolated host cell of  claim 8 , wherein the host cell is a plant cell. 
     
     
         12 . A virus comprising the isolated polynucleotide of  claim 1 . 
     
     
         13 . A process for producing an isolated host cell comprising the chimeric gene of  claim 6 , the process comprising either transforming or transfecting an isolated compatible host cell with the chimeric gene of  claim 6 . 
     
     
         14 . A farnesyltransferase polypeptide of at least 300 amino acids that has at least 80% homology based on the Clustal method of alignment compared to a polypeptide selected from the group consisting of SEQ ID NO:2, 4, 6, 8, 10, 12, 14, 16 and 18. 
     
     
         15 . A method of selecting an isolated polynucleotide that affects the level of expression of a farnesyltransferase polypeptide in a plant cell, the method comprising the steps of:
 constructing an isolated polynucleotide comprising a nucleotide sequence of at least one of 30 contiguous nucleotides derived from a nucleotide sequence selected from the group consisting of SEQ ID NO:1, 3, 5, 7, 9, 11, 13, 15, 17 and the complement of such nucleotide sequences;   introducing the isolated polynucleotide into a plant cell;   measuring the level of farnesyltransferase polypeptide in the plant cell containing the polynucleotide; and   comparing the level of farnesyltransferase polypeptide in the plant cell containing the isolated polynucleotide with the level of farnesyltransferase polypeptide in a plant cell that does not contain the polynucleotide.   
     
     
         16 . The method of  claim 15  wherein the isolated polynucleotide comprises a nucleic acid sequence selected from the group consisting of SEQ ID NO:1, 3, 5, 7, 9, 11, 13, 15, and 17 that codes for the polypeptide selected from the group consisting of SEQ ID NO:2, 4, 6, 8, 10, 12, 14, 16 and 18. 
     
     
         17 . The method of  claim 15  wherein the isolated polynucleotide is DNA. 
     
     
         18 . The method of  claim 15  wherein the isolated polynucleotide is RNA. 
     
     
         19 . The method of  claim 15  wherein the isolated polynucleotide is a chimeric gene comprising the nucleotide sequence operably linked to suitable regulatory sequences. 
     
     
         20 . A method of selecting an isolated polynucleotide that affects the level of expression of farnesyltransferase polypeptide in a plant cell, the method comprising the steps of:
 constructing the isolated polynucleotide of  claim 1 ;   introducing the isolated polynucleotide into a plant cell;   measuring the level of farnesyltransferase polypeptide in the plant cell containing the polynucleotide; and   comparing the level of farnesyltransferase polypeptide in the plant cell containing the isolated polynucleotide with the level of farnesyltransferase polypeptide in a plant cell that does not contain the isolated polynucleotide.   
     
     
         21 . A method of obtaining a nucleic acid fragment encoding a substantial portion of a farnesyltransferase gene comprising the steps of:
 synthesizing an oligonucleotide primer comprising a nucleotide sequence of at least one of 40 contiguous nucleotides derived from a nucleotide sequence selected from the group consisting of SEQ ID NO:1, 3, 5, 7, 9, 11, 13, 15, 17 and the complement of such nucleotide sequences; and   amplifying a nucleic acid sequence using the oligonucleotide primer.   
     
     
         22 . A method of obtaining a nucleic acid fragment encoding all or a substantial portion of the amino acid sequence encoding a farnesyltransferase protein comprising the steps of:
 probing a cDNA or genomic library with an isolated polynucleotide comprising a nucleotide sequence of at least one of 30 contiguous nucleotides derived from a nucleotide sequence selected from the group consisting of SEQ ID NO:1, 3, 5, 7, 9, 11, 13, 15, 17, and the complement of such nucleotide sequences;   identifying a DNA clone that hybridizes with the isolated polynucleotide;   isolating the identified DNA clone; and   sequencing the cDNA or genomic fragment that comprises the isolated DNA clone.   
     
     
         23 . The isolated polynucleotide of  claim 1 , wherein the first polypeptide is compared to the corn farnesyltransferase polypeptide of SEQ ID NO:2. 
     
     
         24 . The isolated polynucleotide of  claim 1 , wherein the first polypeptide is compared to the rice farnesyltransferase polypeptide of SEQ ID NO:4. 
     
     
         25 . The isolated polynucleotide of  claim 1 , wherein the first polypeptide is compared to the soybean farnesyltransferase polypeptide of SEQ ID NO:6. 
     
     
         26 . The isolated polynucleotide of  claim 1 , wherein the first polypeptide is compared to the soybean farnesyltransferase polypeptide of SEQ ID NO:8. 
     
     
         27 . The isolated polynucleotide of  claim 1 , wherein the first polypeptide is compared to the wheat farnesyltransferase polypeptide of SEQ ID NO:10. 
     
     
         28 . The isolated polynucleotide of  claim 1 , wherein the first polypeptide is compared to the corn farnesyltransferase polypeptide of SEQ ID NO:12. 
     
     
         29 . The isolated polynucleotide of  claim 1 , wherein the first polypeptide is compared to the rice farnesyltransferase polypeptide of SEQ ID NO:14. 
     
     
         30 . The isolated polynucleotide of  claim 1 , wherein the first polypeptide is compared to the soybean farnesyltransferase polypeptide of SEQ ID NO:16. 
     
     
         31 . The isolated polynucleotide of  claim 1 , wherein the first polypeptide is compared to the soybean farnesyltransferase polypeptide of SEQ ID NO:18.