Plant diacylglycerol acyltransferases
AssigneeE. I. Du Pont de Nemours and Company
InventorsKarlene H Butler, Edgar Benjamin Cahoon, Rebecca E Cahoon, Anthony J Kinney
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Abstract
This invention relates to an isolated nucleic acid fragment encoding a diacylglycerol acyltransferase. The invention also relates to the construction of a chimeric gene encoding all or a portion of the diacylglycerol acyltransferase, in sense or antisense orientation, wherein expression of the chimeric gene results in production of altered levels of the diacylglycerol acyltransferase in a transformed host cell.
Claims16
An isolated polynucleotide comprising a nucleotide sequence encoding a first polypeptide of at least 50 amino acids that has at least 60% identity based on the Clustal method of alignment when compared to a polypeptide selected from the group consisting of SEQ ID NOs:4, 6, 8, 10, 14, 20 and 22,
- or an isolated polynucleotide comprising the complement of the nucleotide sequence.
A method of selecting an isolated polynucleotide that affects the level of expression of a diacylglycerol acyltransferase polypeptide in a plant cell, the method comprising the steps of:
- (a) 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 NOs:
- 1, 3, 5, 7, 11, 13, 15, 17, 19, 21, and the complement of such nucleotide sequences;
- (b) introducing the isolated polynucleotide into a plant cell;
- (c) measuring the level of a polypeptide in the plant cell containing the polynucleotide;
- and
- (d) comparing the level of polypeptide in the plant cell containing the isolated polynucleotide with the level of polypeptide in a plant cell that does not contain the isolated polynucleotide.
A method for evaluating at least one compound for its ability to inhibit the activity of a diacylglycerol acyltransferase, the method comprising the steps of:
- (a) transforming a host cell with a chimeric gene comprising a nucleic acid fragment encoding a diacylglycerol acyltransferase, operably linked to suitable regulatory sequences;
- (b) growing the transformed host cell under conditions that are suitable for expression of the chimeric gene wherein expression of the chimeric gene results in production of the diacylglycerol acyltransferase encoded by the operably linked nucleic acid fragment in the transformed host cell;
- (c) optionally purifying the diacylglycerol acyltransferase expressed by the transformed host cell;
- (d) treating the diacylglycerol acyltransferase with a compound to be tested;
- and
- (e) comparing the activity of the diacylglycerol acyltransferase that has been treated with a test compound to the activity of an untreated diacylglycerol acyltransferase, thereby selecting compounds with potential for inhibitory activity.
Dependencies are inferred from the claim text.