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 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 pair of primers 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.
2. A pair of polynucleotide primers comprising a first polynucleotide primer and a second polynucleotide primer which function together in the presence of event DP-004114-3 DNA template in a sample to produce an amplicon diagnostic for event DP-004114-3, wherein a detectable label or reporter molecule is attached to the first and/or second polynucleotide primer.
3. The pair of polynucleotide primers according to claim 2 , 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.
4. The pair of polynucleotide primers according to claim 3 , wherein
(a) the first polynucleotide primer comprises at least 10 contiguous nucleotides of a nucleotide sequence selected from the group consisting of nucleotides 1-2422 of SEQ ID NO: 6, nucleotides 14348-16752 of SEQ ID NO: 6, and the full length complements thereof; and
(b) the second polynucleotide primer comprises at least 10 contiguous nucleotides from nucleotides 2423-14347 of SEQ ID NO: 6, or the full length complements thereof.
5. The pair of polynucleotide primers according to claim 4 , 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, SEQ ID NOs: 22-26 and the full length complements thereof; and
(b) the second polynucleotide primer comprises a nucleotide sequence selected from the group consisting of SEQ ID NO: 12, SEQ ID NOs: 14-19, SEQ ID NO: 21, and the full length complements thereof.
6. The primer pair of claim 4 , wherein said first primer and said second primer are at least 18 nucleotides.
7. The primer pair of claim 4 , wherein said first primer and said second primer are at least 24 nucleotides.
8. A kit for detecting nucleic acids that are unique to event DP-004114-3 comprising at least one nucleic acid molecule of sufficient length of contiguous polynucleotides to function as a primer or probe in a nucleic acid detection method, 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.