Plant farnesyltransferases
AssigneesE. I. Du Pont de Nemours and Company, Pioneer Hi-Bred International, Inc.
InventorsRebecca E. Cahoon, Timothy George Helentjaris, Guo-Hua Miao
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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.
Claims31
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.
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.
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.
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.
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.
Dependencies are inferred from the claim text.