Maize event DP-004114-3 and methods for detection thereof
The invention provides DNA compositions that relate to transgenic insect resistant maize plants. Also provided are assays for detecting the presence of the maize DP-004114-3 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 method of determining zygosity of DNA of a corn plant comprising corn event DP-004114-3 in a biological sample comprising:
(a) contacting said sample with a first primer selected from the group consisting of SEQ ID NOs: 11, 13, 23 and 24, and a second primer selected from the group consisting of SEQ ID NOs: 20, 22, 25 and 26, such that
(1) when used in a nucleic acid amplification reaction comprising corn event DP-004114-3 DNA, produces a first amplicon that is diagnostic for corn event DP-004114-3, and
(2) when used in a nucleic acid amplification reaction comprising corn genomic DNA other than DP-004114-3 DNA, produces a second amplicon that is diagnostic for corn genomic DNA other than DP-004114-3 DNA;
(b) performing a nucleic acid amplification reaction; and
(c) detecting the amplicons so produced, wherein detection of presence of both amplicons indicates that said sample is heterozygous for corn event DP-004114-3 DNA, wherein detection of only the first amplicon indicates that said sample is homozygous for corn event DP-004114-3 DNA.
2. A method of detecting the presence of a nucleic acid molecule that is unique to event DP-004114-3 in a sample comprising corn nucleic acids, the method comprising:
(a) contacting the sample with a first primer and a second primer comprising a nucleic acid sequence, wherein the first and second primer are selected from the group consisting of;
i) SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26; or
ii) the complements of SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO:
14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26,
that when used in a nucleic-acid amplification reaction with genomic DNA from event DP-004114-3 produces an amplicon that is diagnostic for event DP-004114-3;
(b) performing a nucleic acid amplification reaction, thereby producing the amplicon; and
(c) detecting the amplicon.
3. A pair of polynucleotide primers, wherein the sequence of the first polynucleotide primer is or is complementary to a corn plant genome sequence flanking the point of insertion of a heterologous DNA sequence inserted into the corn plant genome of event DP-004114-3, and the sequence of the second polynucleotide primer is or is complementary to the heterologous DNA sequence inserted into the genome of event DP-004114-3, wherein
(a) the first polynucleotide primer comprises a nucleotide sequence selected from the group consisting of SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 20, and SEQ ID NOs: 22-26, and the second polynucleotide primer comprises a nucleotide sequence selected from the group consisting of SEQ ID NO: 12, SEQ ID NOs: 14-19, and SEQ ID NO: 21; or
(b) the first polynucleotide primer comprises a nucleotide sequence selected from the group consisting of the complement of SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 20, and SEQ ID NOs: 22-26, and the second polynucleotide primer comprises a nucleotide sequence selected from the group consisting of the complement of SEQ ID NO: 12, SEQ ID NOs: 14-19, and SEQ ID NO: 21, wherein a detectable label or reporter molecule is attached to the first and/or second polynucleotide primer, wherein the detectable label or reporter molecule is not an additional segment of the natural polynucleotide from which the recited SEQ ID NO was derived.
4. A kit for detecting nucleic acids that are unique to event DP-004114-3 comprising:
(a) a first polynucleotide primer comprises a nucleotide sequence selected from the group consisting of SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 20, and SEQ ID NOs: 22-26 and a second polynucleotide primer comprises a nucleotide sequence selected from the group consisting of SEQ ID NO: 12, SEQ ID NOs: 14-19, and SEQ ID NO: 21, or
(b) a first polynucleotide primer comprises a nucleotide sequence selected from the group consisting of the complement of SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 20, SEQ ID NOs: 22-26 and the second polynucleotide primer comprises a nucleotide sequence selected from the group consisting of the complement of SEQ ID NO: 12, SEQ ID NOs: 14-19, and SEQ ID NO: 21, wherein a detectable label or reporter molecule is attached to the first or second primer, wherein the detectable label or reporter molecule is not an additional segment of the natural polynucleotide from which the recited SEQ ID NO was derived, and which upon amplification of or hybridization to a target nucleic acid sequence in a sample followed by detection of the amplicon or hybridization to the target sequence, are diagnostic for the presence of nucleic acid sequences unique to event DP-004114-3 in the sample.