Corn event DAS-59122-7 and methods for detection thereof
View Patent ↗The invention provides DNA compositions that relate to transgenic insect resistant maize plants. Also provided are assays for detecting the presence of the maize DAS-59122-7 event based on the DNA sequence of the recombinant construct inserted into the maize genome and the DNA sequences flanking the insertion site. Kits and conditions useful in conducting the assays are provided.
1. A kit for identifying event DAS-59122-7 in a biological sample which detects a DAS-59122-7 specific region, said kit comprising a primer pair which when used in an amplification reaction is capable of amplifying a DAS-59122-7 specific region, said primer pair comprising a first primer and a second primer, wherein said first primer and said second primer are selected from the group consisting of:
(a) a first primer that is capable of annealing to a nucleic acid molecule comprising the nucleotide sequence set forth in SEQ ID NO: 19 or the full complement thereof and a second primer that is capable of annealing to a nucleic acid molecule comprising the nucleotide sequence set forth in SEQ ID NO: 20 or the full complement thereof;
(b) a first primer that is capable of annealing to a nucleic acid molecule comprising the nucleotide sequence set forth in SEQ ID NO: 19 or the full complement thereof and a second primer that is capable of annealing to a nucleic acid molecule comprising the nucleotide sequence set forth in SEQ ID NO: 24 or the full complement thereof; and
(c) a first primer that is capable of annealing to a nucleic acid molecule comprising the nucleotide sequence set forth in SEQ ID NO: 20 or the full complement thereof and a second primer that is capable of annealing to a nucleic acid molecule comprising the nucleotide sequence set forth in SEQ ID NO: 24 or the full complement thereof.
2. A method for identifying event DAS-59122-7 in a biological sample, comprising detecting a DNA molecule in a biological sample with a probe or first primer which specifically recognizes a sequence within SEQ ID NO: 19, SEQ ID NO: 20, or the full complement of SEQ ID NO: 19 or SEQ ID NO: 20, wherein said DNA molecule comprises at least one junction sequence of DAS-59122-7, thereby identifying event DAS-5912297 in said biological sample.
3. The method of claim 2 , further comprising amplifying a DNA fragment from a nucleic acid present in said biological sample using a polymerase chain reaction with at least two primers, wherein said first primer recognizes a sequence within SEQ ID NO: 19, SEQ ID NO: 20, or the full complement of SEQ ID NO: 19 or SEQ ID NO: 20, and a second primer recognizes a sequence within SEQ ID NO: 20, SEQ ID NO: 24, or the full complement of SEQ ID NO: 20 or SEQ ID NO: 24.
4. The method of claim 3 , wherein said first primer recognizes a sequence within SEQ ID NO: 19 or the full complement thereof, and said second primer recognizes a sequence within SEQ ID NO: 24 or the full complement thereof.
5. The method of claim 3 , wherein said first primer recognizes a sequence within SEQ ID NO: 20 or the full complement thereof, and said second primer recognizes a sequence within SEQ ID NO: 24 or the full complement thereof.
6. The method of claim 4 , wherein said first and second primers comprise the sequence of SEQ ID NO: 18 and SEQ ID NO: 1 respectively.
7. The method of claim 4 , wherein said first and second primers comprise the sequence of SEQ ID NO: 10 and SEQ ID NO:9 respectively.
8. The method of claim 5 , wherein said first and second primers comprise the sequence of SEQ ID NO: 2 and SEQ ID NO: 17 respectively.
9. The method of claim 5 , wherein said first and second primers comprise the sequence of SEQ ID NO: 8 and SEQ ID NO: 17 respectively.
10. The method of claim 4 , wherein said first and second primers comprise the sequence of SEQ ID NO: 36 and SEQ ID NO: 37 respectively.
11. The method of claim 6 , comprising amplifying a fragment of about 555 by using a DAS-59122-7 PCR identification protocol.
12. The method of claim 7 , comprising amplifying a fragment of about 313 by using a DAS-59122-7 PCR identification protocol.
13. The method of claim 8 , comprising amplifying a fragment of about 547 by using a DAS-59122-7 PCR identification protocol.
14. The method of claim 9 , comprising amplifying a fragment of about 754 by using a DAS-59122-7 PCR identification protocol.
15. The method of claim 10 , comprising amplifying a fragment of about 104 by using a DAS-59122-7 PCR identification protocol.
16. A method for confirming seed purity, comprising detecting a DNA molecule in plant material with a specific primer or probe which specifically recognizes a sequence within SEQ ID NO: 19 or SEQ ID NO: 20 or the full complement of SEQ ID NO: 19 or SEQ ID NO: 20, in a seed sample, wherein said DNA molecule comprises at least one junction sequence of DAS-59122-7, thereby confirming the seed sample's purity for event DAS-59122-7.
17. A method for screening seeds for the presence of event DAS-59122-7, comprising detecting a DNA molecule in plant material with a specific primer or probe which specifically recognizes a sequence within SEQ ID NO: 19 or SEQ ID NO: 20 or the full complement of SEQ ID NO: 19 or SEQ ID NO: 20, in a sample of a seed lot, wherein said DNA molecule comprises at least one junction sequence of DAS-59122-7, thereby screening said seeds for the presence of event DAS-59122-7.