CE43-67B insecticidal cotton
View Patent ↗The present application relates an insect resistant transgenic cotton plant. In particular, it relates to a specific event, designated CE43-67B. The application also relates to polynucleotides which are characteristic of the CE43-67B event, plants comprising said polynucleotides, and methods of detecting the CE43-67B event.
1. A method for detecting a plant which contains the polynucleotide depicted as SEQ ID NO: 1, said method comprising:
a) preparing a sample containing the genomic DNA of the plant to be tested;
b) obtaining a pair of primers that, when used in a nucleic acid amplification reaction with genomic DNA from cotton event CE43-67B produce an amplicon comprising at least 18 contiguous nucleotides of the sequence depicted as SEQ ID NO: 3 and the complement thereof;
c) adding said pair of primers to said sample and the means for performing an amplification reaction;
d) performing an amplification reaction; and
e) detecting the thus amplified sequence.
2. The method according to claim 1 , wherein said amplified sequence comprises at least 20 contiguous nucleotides.
3. The method of claim 1 , wherein the pair of primers comprise a first primer sequence which is or is complementary to a cotton plant genome flanking the point of insertion of a heterologous DNA sequence inserted into the cotton plant genome of cotton event CE43-678, and a second primer sequence which is or is complementary to the heterologous DNA sequence inserted into the cotton plant genome of the cotton event CE43-67B.
4. The method of claim 3 , wherein the first primer sequence comprises a sequence selected from the group consisting of SEQ ID NO: 5, SEQ ID NO: 11, SEQ ID NO: 17, and SEQ ID NO: 19; and wherein the second primer sequence comprises a sequence selected from the group consisting of SEQ ID NO: 6 and SEQ ID NO: 12.
5. The method of claim 4 , wherein the first primer sequence comprises SEQ ID NO: 5 and the second primer sequence comprises SEQ ID NO: 6.
6. The method of claim 4 , wherein the first primer sequence comprises SEQ ID NO: 11 and the second primer sequence comprises SEQ ID NO: 12.
7. A method for detecting a plant which contains the polynucleotide depicted as SEQ ID NO: 2, said method comprising:
a) preparing a sample containing the genomic DNA of the plant to be tested;
b) obtaining a pair of primers that, when used in a nucleic acid amplification reaction with genomic DNA from cotton event CE43-67B produce an amplicon comprising at least 18 contiguous nucleotides of the sequence depicted as SEQ ID NO: 4 and the complement thereof;
c) adding said pair of primers to said sample and the means for performing an amplification reaction;
d) performing an amplification reaction; and
e) detecting the thus amplified sequence.
8. The method according to claim 7 , wherein said sequence comprises at least 20 contiguous nucleotides.
9. The method of claim 7 , wherein the pair of primers comprise a first primer sequence which is or is complementary to a cotton plant genome flanking the point of insertion of a heterologous DNA sequence inserted into the cotton plant genome of cotton event CE43-67 B, and a second primer sequence which is or is complementary to, the heterologous DNA sequence inserted into the cotton plant genome of the cotton event CE43-67B.
10. The method of claim 9 , wherein the first primer sequence comprises a sequence selected from the group consisting of SEQ ID NO: 9 and SEQ ID NO: 15; and wherein the second primer sequence comprises a sequence selected from the group consisting of SEQ ID NO: 10, SEQ ID NO: 14, and SEQ ID NO: 16.
11. The method of claim 10 , wherein the first primer sequence comprises SEQ ID NO: 9 and the second primer sequence comprises SEQ ID NO: 10.
12. A method for detecting a plant which contains the polynucleotide depicted as SEQ ID NO: 1 and/or the polynucleotide depicted as SEQ ID NO: 2 said method comprising:
a) preparing a sample containing the genomic DNA of the plant to be tested;
b) obtaining at least one probe which is capable of hybridizing to a sequence selected from the group consisting of a sequence comprising at least 1 8 contiguous nucleotides of the sequence depicted as SEQ ID NO: 3 and a sequence comprising at least 18 contiguous nucleotides of the sequence depicted as SEQ ID NO: 4;
c) adding at least one of the probes of step (b) to said sample under conditions which allow said probe to hybridize with a complementary nucleic acid within said sample;
d) removing substantially non-hybridized probe; and
e) detecting the thus hybridized probe to identify if the sample contains said polynucleotide.
13. The method according to claim 12 wherein said probe comprises a sequence selected from the group consisting of SEQ ID NO: 3, SEQ ID NO: 4, and SEQ ID•NO: 15.
14. The method according to claim 12 wherein said substantially non-hybridized probe is removed by rinsing said probe under high stringency conditions.