Brassica events LBFLFK and LBFDAU and methods for detection thereof

Patent metadata

Field	Value
Publication number	US-11813302-B2
Application number	US-202117346522-A
Country	US
Kind code	B2
Filing date	Jun 14, 2021
Priority date	Nov 14, 2014
Publication date	Nov 14, 2023
Grant date	Nov 14, 2023

Abstract

The present invention provides transgenic Brassica events LBFLFK and LBFDAU and progeny thereof, and cells, seeds, plants comprising DNA diagnostic for these events. The invention also provides artificial oligonucleotide primers and probes that are diagnostic for the LBFLFK and LBFDAU events and their progeny in a sample, and methods for detecting the presence of the LBFLFK and LBFDAU events and their progeny in a sample. The invention further provides oil and commodity products derived from the LBFLFK and LBFDAU events.

First claim

The invention claimed is:

1. A method of detecting the presence of DNA corresponding to Brassica event LBFLFK in a sample comprising DNA, the method comprising the steps of:
(a) contacting the sample with an LBFLFK Locus 1 primer pair that, when used in a nucleic acid amplification reaction with genomic DNA from Brassica event LBFLFK, produces a Locus 1 amplicon that is diagnostic for Brassica event LBFLFK;
(b) performing a nucleic acid amplification reaction, thereby producing the Locus 1 amplicon; and
(c) detecting the amplicon, wherein the amplicon comprises the LBFLFK Locus 1 junction region SEQ ID NO:4 or SEQ ID NO:5, or the complement thereof.

2. The method of claim 1 , wherein the LBFLFK Locus 1 primer pair comprises:
(i) a first primer selected from the group consisting of at least 11 consecutive nucleotides of SEQ ID NO:6, at least 11 consecutive nucleotides of the complement of SEQ ID NO:6, at least 11 consecutive nucleotides of SEQ ID NO:7, and at least 11 consecutive nucleotides of the complement of SEQ ID NO:7; and
(ii) a second primer selected from the group consisting of at least 11 consecutive nucleotides of SEQ ID NO:3 and at least 11 consecutive nucleotides of the complement of SEQ ID NO:3.

3. The method of claim 2 , wherein the first primer comprises SEQ ID NO:8 and the second primer comprises SEQ ID NO:9.

4. A method of detecting the presence of DNA corresponding to Brassica event LBFLFK in a sample comprising DNA, the method comprising the steps of:
(a) contacting the sample with an LBFLFK Locus 2 primer pair that, when used in a nucleic acid amplification reaction with genomic DNA from Brassica event LBFLFK, produces a Locus 2 amplicon that is diagnostic for Brassica event LBFLFK;
(b) performing a nucleic acid amplification reaction, thereby producing the Locus 2 amplicon; and
(c) detecting the amplicon, wherein the amplicon comprises the LBFLFK Locus 2 junction region SEQ ID NO:13 or SEQ ID NO:14, or the complement thereof.

5. The method of claim 4 , wherein the LBFLFK Locus 2 primer pair comprises:
(i) a first primer selected from the group consisting of at least 11 consecutive nucleotides of SEQ ID NO:15, at least 11 consecutive nucleotides of the complement of SEQ ID NO:15, at least 11 consecutive nucleotides of SEQ ID NO:16, and at least 11 consecutive nucleotides of the complement of SEQ ID NO:16; and
(ii) a second primer selected from the group consisting of at least 11 consecutive nucleotides of SEQ ID NO:12 and at least 11 consecutive nucleotides of the complement of SEQ ID NO:12.

6. The method of claim 5 , wherein the first primer comprises SEQ ID NO:17 and the second primer comprises SEQ ID NO:18.

Assignees

• Basf Plant Science Co Gmbh

Inventors

• Senger Toralf
• Marty Laurent
• Kunze Irene
• Rein Dietrich
• Bauer Jörg
• Andre Carl

Classifications

• A61K36/31 Primary

  Brassicaceae or Cruciferae (Mustard family), e.g. broccoli, cabbage or kohlrabi · CPC title

• C12N15/8247 Primary

  involving modified lipid metabolism, e.g. seed oil composition · CPC title

• C12Q1/6895

  for plants, fungi or algae · CPC title

• A61K31/202

  having three or more double bonds, e.g. linolenic (eicosanoids, e.g. leukotrienes A61K31/557) · CPC title

• C12N9/0071

  acting on paired donors with incorporation of molecular oxygen (1.14) · CPC title