Back to results
US 9,006,514 B2Granted
Plant membrane O-acyl transferase (MBOAT) family protein sequences and their uses for altering fatty acid compositions
AssigneeE. I. Du Pont de Nemours and Company
InventorsHoward Glenn Damude, Knut Meyer
Life of this patent
Estimating the term…
Abstract
This invention is in the field of biotechnology, in particular, this pertains to polynucleotide sequences encoding membrane bound O-acyltransferase genes and the use of these acyltransferases for altering fatty acid profiles in oilseed plants. Methods for increasing elongation and desaturation conversion efficiencies are also disclosed.
Claims8
A method for increasing the content of at least one unusual fatty acid in an oilseed plant cell comprising:
- (a) transforming the oilseed plant cell with:
- (i) an isolated polynucleotide comprising:
- (1) a nucleotide sequence encoding a polypeptide with membrane-bound O-acyltransferase (MBOAT) activity, wherein, based on the Clustal V method of alignment with pairwise alignment default parameters of KTUPLE=1, GAP PENALTY=3, WINDOW=5 and DIAGONALS SAVED=5, the polypeptide has an amino acid sequence of at least 85% sequence identity when compared to SEQ ID NO:122;
- or
- (2) the full complement of the nucleotide sequence of (1);
- and a heterologous promoter operably linked to either (1) or (2);
- and
- (ii) at least one additional recombinant construct comprising an isolated polynucleotide, operably linked to at least one regulatory sequence, encoding a polypeptide selected from the group consisting of a delta-4 desaturase, a delta-5 desaturase, a delta-6 desaturase, a delta-8 desaturase, a delta-12 desaturase, a delta-15 desaturase, a delta-17 desaturase, a delta-9 desaturase, a delta-9 elongase, a C14/16 elongase, a C16/18 elongase, a C18/20 elongase, a C20/22 elongase, a DGAT, an MBOAT, a fatty acid conjugase, a hydroxylase, an acetylenase, an epoxidase and a multizyme;
- (b) regenerating an oilseed plant from the transformed cell of step (a);
- and
- (c) selecting those seeds obtained from the plants of step (b) having an increased level of at least one unusual fatty acid selected from the group consisting of GLA, STA, EDA, ERA, DGLA, ETA, ARA, and EPA;
- when compared to the level in seeds obtained from a transgenic plant comprising at least one recombinant construct comprising an isolated polynucleotide, operably linked to at least one regulatory sequence, encoding the same polypeptide as encoded by the isolated polynucleotide of step (a)(ii), but not comprising the polynucleotide of step (a)(i).
9 more elements
A method for increasing at least one conversion efficiency, wherein said conversion efficiency is at least one selected from the group consisting of:
- C18 to C20 elongation, delta-6 desaturation, delta-9 elongation, delta-8 desaturation, conjugation to Eleostearic acid, and conjugation to Calendic acid, in an oilseed plant cell comprising:
- (a) transforming the oilseed plant cell with:
- (i) an isolated polynucleotide comprising:
- (1) a nucleotide sequence encoding a polypeptide with membrane-bound O-acyltransferase (MBOAT) activity, wherein, based on the Clustal V method of alignment with pairwise alignment default parameters of KTUPLE=1, GAP PENALTY=3, WINDOW=5 and DIAGONALS SAVED=5, the polypeptide has an amino acid sequence of at least 85% sequence identity when compared to SEQ ID NO:122;
- or
- (2) the full complement of the nucleotide sequence of (1);
- and a heterologous promoter operably linked to either (1) or (2);
- and
- (ii) at least one additional recombinant construct comprising an isolated polynucleotide, operably linked to at least one regulatory sequence, encoding a polypeptide selected from the group consisting of a delta-4 desaturase, a delta-5 desaturase, a delta-6 desaturase, a delta-8 desaturase, a delta-12 desaturase, a delta-15 desaturase, a delta-17 desaturase, a delta-9 desaturase, a delta-9 elongase, a C14/16 elongase, a C16/18 elongase, a C18/20 elongase, a C20/22 elongase, a DGAT, an MBOAT, a fatty acid conjugase, a hydroxylase, an acetylenase, an epoxidase and a multizyme;
- (b) regenerating an oilseed plant from the transformed cell of step (a);
- and
- (c) selecting those seeds obtained from the plants of step (b) having an increased C18 to C20 elongation conversion efficiency and an increased delta-6 desaturation conversion efficiency when compared to the level in seeds obtained from a transgenic plant comprising at least one recombinant construct comprising an isolated polynucleotide, operably linked to at least one regulatory sequence, encoding the same polypeptide as encoded by the isolated polynucleotide of step (a)(ii), but not comprising the polynucleotide of step (a)(i).
9 more elements
An oil seed plant or oil seed comprising:
- (a) an isolated polynucleotide comprising:
- (i) a nucleotide sequence encoding a polypeptide with membrane-bound O-acyltransferase (MBOAT) activity, wherein, based on the Clustal V method of alignment with pairwise alignment default parameters of KTUPLE=1, GAP PENALTY=3, WINDOW=5 and DIAGONALS SAVED=5, the polypeptide has an amino acid sequence of at least 85% sequence identity when compared to SEQ ID NO:122;
- or
- (ii) the full complement of the nucleotide sequence of (i);
- and a heterologous promoter operably linked to either (i) or (ii);
- and
- (b) at least one additional recombinant construct comprising an isolated polynucleotide, operably linked to at least one regulatory sequence, encoding a polypeptide selected from the group consisting of a delta-4 desaturase, a delta-5 desaturase, a delta-6 desaturase, a delta-8 desaturase, a delta-12 desaturase, a delta-15 desaturase, a delta-17 desaturase, a delta-9 desaturase, a delta-9 elongase, a C14/16 elongase, a C16/18 elongase, a C18/20 elongase, a C20/22 elongase, a DGAT, an MBOAT, a fatty acid conjugase, a hydroxylase, an acetylenase, an epoxidase, and a multizyme.
4 more elements
Dependencies are inferred from the claim text.