Plant caffeic acid-3-O-methyltransferase homologs
AssigneeE. I. Du Pont de Nemours and Company
InventorsRebecca E. Cahoon, J. Antoni Rafalski, Jennie B. Shen
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Abstract
This invention relates to an isolated nucleic acid fragment encoding a protein involved in phenylpropanoid metabolism. The invention also relates to the construction of a chimeric gene encoding all or a portion of the protein involved in phenylpropanoid metabolism, in sense or antisense orientation, wherein expression of the chimeric gene results in production of altered levels of the protein involved in phenylpropanoid metabolism in a transformed host cell.
Claims23
An isolated polynucleotide comprising a first nucleotide sequence encoding a polypeptide of at least 305 amino acids that has at least 92% identity based on the Clustal method of alignment when compared to a polypeptide selected from the group consisting of SEQ ID NOs:2, 4, 6, 8, 10, 12, 14 and 16,
- or a second nucleotide sequence comprising the complement of the first nucleotide sequence.
A polypeptide of at least 305 amino acids that has at least 92% identity based on the Clustal method of alignment when compared to a polypeptide selected from the group consisting of SEQ ID NOs:2, 4, 6, 8, 10, 12, 14 and 16.
A method of obtaining a nucleic acid fragment encoding a phenylpropanoid metabolism polypeptide comprising the steps of:
- (a) synthesizing an oligonucleotide primer 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 NOs:1, 3, 5, 7, 9, 11, 13 and 15 and the complement of such nucleotide sequences;
- and
- (b) amplifying a nucleic acid sequence using the oligonucleotide primer.
A method of obtaining a nucleic acid fragment encoding a phenylpropanoid metabolism polypeptide comprising the steps of:
- (a) probing a cDNA or genomic library with an isolated polynucleotide comprising at least one of 30 contiguous nucleotides derived from a nucleotide sequence selected from the group consisting of SEQ ID NOs:1, 3, 5, 7, 9, 11, 13 and 15 and the complement of such nucleotide sequences;
- (b) identifying a DNA clone that hybridizes with the isolated polynucleotide;
- (c) isolating the identified DNA clone;
- and
- (d) sequencing the cDNA or genomic fragment that comprises the isolated DNA clone.
An isolated polynucleotide comprising the nucleotide sequence having at least one of 30 contiguous nucleotides derived from a nucleic acid sequence selected from the group consisting of SEQ ID NOs:1, 3, 5, 7, 9, 11, and 13 and the complement of such sequences.
A method for positive selection of a transformed cell comprising:
- (a) transforming a host cell with the chimeric gene of claim 5 or an expression cassette of claim 20;
- and
- (b) growing the transformed host cell under conditions which allow expression of the polynucleotide in an amount sufficient to complement a null mutant and alter methylation of both caffeic acid and 5-hydroxyferulic acid to provide a positive selection means.
The method of any of claims 21 or 23 wherein the plant cell is a monocot.
The method of any of claims 21 or 23 wherein the plant cell is a dicot.
An isolated polynucleotide comprising a first nucleotide sequence 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 selected from the group consisting of SEQ ID NOs:18, 20, 22, 24, 26 and 28
- or a second nucleotide sequence comprising the complement of the first nucleotide sequence.
Dependencies are inferred from the claim text.