Corn event DAS-59122-7 and methods for detection thereof
First Claim
Patent Images
1. An isolated DNA molecule comprising a nucleotide sequence selected from the group consisting of:
- a) the nucleotide sequence set forth in SEQ ID NO;
19;
b) the nucleotide sequence set forth in SEQ ID NO;
20;
c) the nucleotide sequence set forth in SEQ ID NO;
23;
d) the nucleotide sequence set forth in SEQ ID NO;
21; and
e) the nucleotide sequence set forth in SEQ ID NO;
22.
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Abstract
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.
45 Citations
49 Claims
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1. An isolated DNA molecule comprising a nucleotide sequence selected from the group consisting of:
-
a) the nucleotide sequence set forth in SEQ ID NO;
19;
b) the nucleotide sequence set forth in SEQ ID NO;
20;
c) the nucleotide sequence set forth in SEQ ID NO;
23;
d) the nucleotide sequence set forth in SEQ ID NO;
21; and
e) the nucleotide sequence set forth in SEQ ID NO;
22.
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2. A kit for identifying event DAS-59122-7 in a biological sample which detects a DAS-59122-7 specific region, said kit comprising at least a first primer, which recognizes a sequence within SEQ ID NO:
- 19 or within SEQ ID NO;
20. - View Dependent Claims (3, 4, 5)
- 19 or within SEQ ID NO;
-
6. A DNA detection kit specific for junction DNA of maize event DAS-59122-7 and its progeny comprising at least one DNA molecule of a sufficient length of contiguous DNA polynucleotides to function in a DNA detection method, that is homologous or complementary to a sequence selected from the group consisting of:
-
a) the nucleotide sequence set forth in SEQ ID NO;
21; and
b) the nucleotide sequence set forth in SEQ ID NO;
22.
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7. A kit for identifying event DAS-59122-7 in a biological sample, said kit comprising a specific probe comprising a sequence which hybridizes with sequences selected from the group consisting of:
-
a) the sequences of SEQ ID NO;
19 and SEQ ID NO;
24; and
b) the sequences of SEQ ID NO;
20 and SEQ ID NO;
24;
contiguous therewith.
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8. A DNA detection kit comprising at least one DNA molecule of a sufficient length of contiguous nucleotides homologous or complementary to SEQ ID NO:
- 21 or SEQ ID NO;
22 that functions as a DNA primer or probe specific for maize event DAS-59122-7 and its progeny.
- 21 or SEQ ID NO;
-
9. A DNA construct comprising:
- a first, second, and third expression cassette, wherein said first expression cassette in operable linkage comprises;
(a) a maize ubiquitin promoter;
(b) a 5′
untranslated exon of a maize ubiquitin gene;
(c) a maize ubiquitin first intron;
(d) a Cry34Ab1 encoding DNA molecule; and
(e) a PinII transcriptional terminator;
said second expression cassette comprising in operable linkage(i) a wheat peroxidase promoter;
(ii) a Cry35Ab1 encoding DNA molecule; and
(iii) a PinII transcriptional terminator; and
said third expression cassette comprising in operable linkage(1) a CaMV 35S promoter;
(2) a pat encoding DNA molecule; and
(3) a 3′
transcriptional terminator from (CaMV) 35S. - View Dependent Claims (10, 11)
- a first, second, and third expression cassette, wherein said first expression cassette in operable linkage comprises;
-
12. A method for identifying event DAS-59122-7 in a biological sample, comprising detecting a DAS-59122-7 specific region with a probe or first primer which specifically recognizes a sequence within SEQ ID NO:
- 19 or SEQ ID NO;
20. - View Dependent Claims (13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25)
- 19 or SEQ ID NO;
-
26. A method of detecting the presence of maize event DAS-59122-7 or progeny thereof in a biological sample, comprising:
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(a) extracting a DNA sample from said biological sample;
(b) providing a pair of DNA primer molecules selected from the group consisting of;
i) the sequences of SEQ ID NO;
18 and SEQ ID NO;
1;
ii) the sequences of SEQ ID NO;
10 and SEQ ID NO;
9;
iii) the sequences of SEQ ID NO;
2 and SEQ ID NO;
17; and
iv) the sequences of SEQ ID NO;
8 and SEQ ID NO;
17;
(c) providing DNA amplification reaction conditions;
(d) performing said DNA amplification reaction, thereby producing a DNA amplicon molecule; and
(e) detecting said DNA amplicon molecule, wherein the detection of said DNA amplicon molecule in said DNA amplification reaction indicates the presence of maize event DAS-59122-7. - View Dependent Claims (27)
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28. A method of detecting the presence of DNA corresponding to the DAS-59122-7 event in a sample, the method comprising:
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(a) contacting the sample comprising maize DNA with a polynucleotide probe that hybridizes under stringent hybridization conditions with DNA from maize event DAS-59122-7 and does not hybridize under said stringent hybridization conditions with a non-DAS-59122-7 maize plant DNA;
(b) subjecting the sample and probe to stringent hybridization conditions; and
(c) detecting hybridization of the probe to the DNA, wherein detection of hybridization indicates the presence of the DAS-59122-7 event.
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29. An isolated DNA nucleotide primer sequence comprising a sequence selected from the group consisting of:
- SEQ ID NOs;
1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 36 and 37, or its complement. - View Dependent Claims (30)
- SEQ ID NOs;
-
31. A pair of DNA molecules comprising:
- a first DNA molecule and a second DNA molecule, wherein the DNA molecules are of a sufficient length of contiguous nucleotides of a sequence selected from the group consisting of;
a) the sequence set forth in SEQ ID NO;
21 or its complement; and
b) the sequence set forth in SEQ ID NO;
22 or its complement;
to function as DNA primers or probes diagnostic for DNA extracted from a DAS-59122-7 corn plant or progeny thereof.
- a first DNA molecule and a second DNA molecule, wherein the DNA molecules are of a sufficient length of contiguous nucleotides of a sequence selected from the group consisting of;
-
32. An isolated DNA molecule comprising a junction sequence comprising a sequence selected from the group consisting of SEQ ID NO:
- 32, 33, 34, and 35 and complements thereof.
-
33. A method for confirming seed purity, comprising detection of a DAS-59122-7 specific region with a specific primer or probe which specifically recognizes a sequence within SEQ ID NO:
- 19 or SEQ ID NO;
20, in a seed sample.
- 19 or SEQ ID NO;
-
34. A method for screening seeds for the presence of event DAS-59122-7, comprising detection of a DAS-59122-7 specific region with a specific primer or probe which specifically recognizes a sequence within SEQ ID NO:
- 19 or SEQ ID NO;
20 in a sample of a seed lot.
- 19 or SEQ ID NO;
-
35. An insect resistant corn plant, or parts thereof, wherein DNA having at least one nucleotide sequence selected from the group consisting of SEQ ID NO:
- 32, 33, 34, and 35 and complements thereof forms part of the plant'"'"'s genome.
- View Dependent Claims (36, 37, 38)
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39. An isolated DNA sequence comprising at least one nucleotide sequence selected from the group consisting of SEQ ID NO:
- 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 36 and 37, or complements thereof.
- 40. A pair of isolated DNA sequences, each comprising at least ten nucleotides and which when used together in a DNA amplification procedure will produce an amplicon diagnostic for event DAS-59122-7.
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42. A method of detecting the presence of the DAS-59122-7 event insertion in corn tissue comprising:
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(a) selecting a primer pair each comprising at least ten nucleotides from SEQ ID NO;
21 or SEQ ID NO;
22 wherein each member of the pair is on opposite sides of a sequence diagnostic for said DAS-59122-7 event insertion;
(b) contacting a sample of said corn tissue with said primer pair;
(c) performing DNA amplification and analyzing for an amplicons. - View Dependent Claims (43)
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44. A method of detecting the presence of the DAS-59122-7 event insertion in corn tissue comprising:
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(a) contacting a sample of said corn tissue with a polynucleotide probe that hybridizes under stringent hybridization conditions with one or more DNA sequence selected from the group consisting of SEQ ID NO;
32, 33, 34, and 35 and complements thereof;
(b) subjecting said sample and probe to stringent hybridization conditions; and
(c) analyzing for hybridization of the probe.
-
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45. A DNA detection kit comprising a polynucleotide probe that hybridizes under stringent hybridization conditions with one or more DNA sequences selected from the group consisting of SEQ ID NO:
- 32, 33, 34, and 35 and complements thereof.
-
46. A DNA detection kit comprising a primer pair each comprising at least 10 nucleotides from within SEQ ID NO:
- 21 and SEQ ID NO;
22, wherein each is on opposite sides of a sequence diagnostic for the DAS-59122-7 event insertion. - View Dependent Claims (47)
- 21 and SEQ ID NO;
-
48. A kit for identifying event DAS-59122-7 in a biological sample which detects a DAS-59122-7 specific region within SEQ ID NO:
- 23.
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49. A method for identifying DAS-59122-7 in a biological sample which detects a DAS-59122-7 specific region within SEQ ID NO:
- 23.
Specification