T227-1 flanking sequence
A herbicide resistant transformed sugar beet that is detectable by the specific primers developed to match the DNA sequences that flank the left and/or right border region of the inserted transgenic DNA and the method of identifying primer pairs containing plant genomic DNA/plasmid DNA. More specifically the present invention covers a specific glyphosate resistant sugar beet plant having an insertion of the transgenic material identified as the T227-1 event. The present invention additionally covers primer pairs: plant genomic DNA/Plasmid DNA that are herein identified. Additionally, these primer pairs for either the left or the right flanking regions make an event specific test for the T227-1 insert of transgenic material.
1. A pair of primers selected from the group consisting of:
the pair of primers identified as 98G94, which consists of nucleotides 1–20 of SEQ ID NO: 15 and 98K86, which consists of nucleotides 730–753 of SEQ ID NO: 10 and;
the pair of primers identified as 98I50, which consists of nucleotides 455–476 of SEQ ID NO: 14 and 98K89, which consists of nucleotides 155–180 of SEQ ID NO: 4.
2. A pair of primers of claim 1 that hybridizes to a transgenic insert of DNA of a transgenic glyphosate-resistant sugar beet, wherein the transgenic insert of DNA comprises at least one right border region, and at least one left border region, and comprises a figwort mosaic virus 35S promoter, DNA encoding a chloroplast transit peptide from the EPSPS gene of Arabidopsis , and an optimized CP4 EPSPS gene from Agrobacteriurn sp. CP4, and a terminator.
3. A pair of primers according to claim 2 comprising DNA which hybridizes to DNA that lies in the right border region of the insert DNA.
4. A pair of primers according to claim 2 comprising DNA which hybridizes to DNA that lies in the left border region of the insert DNA.
5. A pair of primers according to claim 3 or claim 4 , wherein one primer of the pair comprises DNA which hybridizes to genomic DNA of the transgenic beet and the other primer hybridizes to the transgenic insert.
6. A method of detecting a transgenic sugar beet having an insert DNA comprising a figwort mosaic virus 35s promoter, a chioroplast transit peptide from the EPSPS gene of Arabidopsis , an optimized CP4 EPSPS gene from Agrobacterium sp. strain CP4 and a terminator; the method comprising the steps of:
1) amplifying DNA from the transgenic beet in the presence of a pair of primers selected from the group consisting of:
the pair of primers identified as 98G94, which consists of nucleotides 1–20 of SEQ ID NO: 15 and 98K86, which consists of nucleotides 730–753 of SEQ ID NO: 10 and;
the pair of primers identified as 98I50, which consists of nucleotides 455–476 of SEQ ID NO: 14 and 98K89, which consists of nucleotides 155–180 of SEQ ID NO: 4
using the polyrnerase chain reaction and
2) detecting the presence or the absence of the DNA fragment.