# US 2014/0154720 A1 · Plant diacylglycerol acyltransferases > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2014/0154720 A1 - Document ID: US20140154720A1 - Status: Published application - Kind: A1 - Application number: 14/174,014 - Filed: 2014-02-06 - Published: 2014-06-05 - Earliest claimed: 1998-12-02 - Assignees: E. I. Du Pont de Nemours and Company - Applicants: E I Du Pont De Nemours and Company - Inventors: Karlene H Butler; Edgar Benjamin Cahoon; Rebecca E Cahoon; Anthony J Kinney - Web page: https://patentgrep.com/patent/US20140154720A1 ## CPC - C12N 9/1029: {transferring groups other than amino-acyl groups (2.3.1)} - C07K 2319/00: Fusion polypeptide - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) ## 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. ## Claims ### Claim 1 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. ### Claim 2 (depends on 1) The isolated polynucleotide of claim 1, wherein the isolated nucleotide sequence consists of a nucleic acid sequence selected from the group consisting of SEQ ID NOs:1, 7, 13, 15, and 21 that codes for the polypeptide selected from the group consisting of SEQ ID NOs:2, 8, 14, 16, and 22. ### Claim 3 (depends on 1) The isolated polynucleotide of claim 1 wherein the isolated polynucleotide is DNA. ### Claim 4 (depends on 1) The isolated polynucleotide of claim 1 wherein the isolated polynucleotide is RNA. ### Claim 5 (depends on 1) A chimeric gene comprising the isolated polynucleotide of claim 1 operably linked to suitable regulatory sequences. ### Claim 6 (depends on 5) An isolated host cell comprising the chimeric gene of claim 5. ### Claim 7 (depends on 1) An isolated host cell comprising the isolated polynucleotide of claim 1. ### Claim 8 (depends on 7) The isolated host cell of claim 7 wherein the isolated host is selected from the group consisting of yeast, bacteria, plant, and virus. ### Claim 9 (depends on 1) A virus comprising the isolated polynucleotide of claim 1. ### Claim 10 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. ### Claim 11 (depends on 10) The method of claim 10 wherein the isolated polynucleotide consists of a nucleotide sequence selected from the group consisting of SEQ ID NOs:1, 3, 5, 7, 11, 13, 15, 17, 19 and 21 that codes for the polypeptide selected from the group consisting of SEQ ID NOs:2, 4, 6, 8, 10, 12, 14, 16, 18, 20 and 22. ### Claim 12 (depends on 1) 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 the isolated polynucleotide of claim 1; (b) introducing the isolated polynucleotide into a plant cell; and (c) measuring the level of diacylglycerol acyltransferase polypeptide in the plant cell containing the polynucleotide. ### Claim 13 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. ### Claim 14 (depends on 1) A composition comprising the isolated polynucleotide of claim 1. ### Claim 15 (depends on 1) An expression cassette comprising the isolated polynucleotide of claim 1 operably linked to a promoter. ### Claim 16 (depends on 15) A method for positive selection of a transformed cell comprising: (a) transforming a plant cell with the expression cassette of claim 15; and (b) growing the transformed plant cell under conditions allowing expression of the polynucleotide in an amount sufficient to modify oil content in the plant cell to provide a positive selection means.