AAD-1 event DAS-40278-9, related transgenic corn lines, and event-specific identification thereof
View Patent ↗This invention relates in part to plant breeding and herbicide tolerant plants. This invention includes a novel aad-1 transformation event in corn plants comprising a polynucleotide sequence, as described herein, inserted into a specific site within the genome of a corn cell. In some embodiments, said event/polynucleotide sequence can be “stacked” with other traits, including, for example, other herbicide tolerance gene(s) and/or insect-inhibitory proteins. Additionally, the subject invention provides assays for detecting the presence of the subject event in a sample (of corn grain, for example). The assays can be based on the DNA sequence of the recombinant construct, inserted into the corn genome, and on the genomic sequences flanking the insertion site. Kits and conditions useful in conducting the assays are also provided.
1. A method of detecting corn event DAS-40278-9 in a sample comprising corn DNA, wherein representative seed comprising said event has been deposited with the American Type Culture Collection (ATCC) under Accession No. PTA-10244, wherein said method comprises contacting said sample with at least one polynucleotide that is diagnostic for said event, wherein said polynucleotide comprises nucleotides 6679 to 6700 of SEQ ID NO: 29 or full complement thereof.
2. A method of detecting corn event DAS-40278-9 in a sample comprising corn DNA, wherein said method comprises contacting said sample with
(a) a first primer that binds to a flanking sequence of the event, said flanking sequence selected from the group consisting of nucleotides 1-1873 of SEQ ID NO: 29, nucleotides 6690-8557 of SEQ ID NO: 29, and full complements thereof; and
(b) a second primer that binds to the insert sequence of the event, said insert sequence comprising nucleotides 1874-6689 of SEQ ID NO: 29 or full complement thereof;
subjecting said sample to polymerase chain reaction; and
assaying for an amplicon generated between said primers.
3. The method of claim 2 , wherein said first primer is selected from the group consisting of SEQ ID NO: 5, 15, 16, 21, 24, and 26; and wherein said second primer is selected from the group consisting of SEQ ID NO: 6, 17, 18, 23, 25, and 27.
4. The method of claim 1 , wherein said polynucleotide comprises at least 30 contiguous nucleotides and hybridizes under stringent conditions with a sequence selected from the group consisting of nucleotides 6679 to 6700 of SEQ ID NO: 29 and full complement thereof, and does not hybridize under stringent conditions with a sequence not comprising nucleotides 6679 to 6700 of SEQ ID NO: 29 or full complement thereof; wherein said method further comprises:
subjecting said sample and said polynucleotide to stringent hybridization conditions; and
assaying said sample for hybridization of said polynucleotide to said DNA.
5. The method of claim 1 , further comprising contacting said sample with an additional polynucleotide, wherein said additional polynucleotide comprises nucleotides 1863 to 1875 of SEQ ID NO: 29 or full complement thereof.
6. The method of claim 4 , further comprising contacting said sample with an additional polynucleotide, wherein said additional polynucleotide comprises at least 30 contiguous nucleotides and hybridizes under stringent conditions with a sequence selected from the group consisting of nucleotides 1863 to 1875 of SEQ ID NO: 29 and full complement thereof, and does not hybridize under stringent conditions with a sequence not comprising nucleotides 1863 to 1875 of SEQ ID NO: 29 or full complement thereof; wherein said method further comprises:
subjecting said sample and said additional polynucleotide to stringent hybridization conditions; and
assaying said sample for hybridization of said additional polynucleotide to said DNA.