Corn plant and seed corresponding to transgenic event MON89034 and methods for detection and use thereof

• US 8,062,840 B2
• Filed: 05/25/2007
• Issued: 11/22/2011
• Est. Priority Date: 05/26/2006
• Status: Active Grant

• Current Assignee: Monsanto Technology LLC (Bayer AG)
• Original Assignee: Monsanto Technology LLC (Bayer AG)
• Inventors: Allen, Jennifer R., Kelly, Rebecca A., Rice, James F., Anderson, Heather M., Groat, Jeanna R., Korte, John, Johnson, Scott C.
• Primary Examiner(s): Horlick; Kenneth R.
• Assistant Examiner(s): Tung; Joyce
• Application Number:
  US11/753,574
• Publication Number:
  US 20080260932A1
• Time in Patent Office: 1,642 Days
• Field of Search: 536/22.1, 435/6, 435/91.2, 435/6.12
• US Class Current: 435/6
• CPC Class Codes: C07K 14/325 Bacillus thuringiensis crystal peptides, i.e. delta-endotoxins
• CPC Class Codes: C12N 15/8286 for insect resistance
• CPC Class Codes: C12Q 1/6895 for plants, fungi or algae
• CPC Class Codes: C12Q 2600/158 Expression markers
• CPC Class Codes: Y02A 40/146 Genetically Modified [GMO] plants, e.g. transgenic plants

Abstract

The present invention provides a transgenic corn event MON89034, and cells, seeds, and plants comprising DNA diagnostic for the corn event. The invention also provides compositions comprising nucleotide sequences that are diagnostic for said corn event in a sample, methods for detecting the presence of said corn event nucleotide sequences in a sample, probes and primers for use in detecting nucleotide sequences that are diagnostic for the presence of said corn event in a sample, growing the seeds of such corn event into corn plants, and breeding to produce corn plants comprising DNA diagnostic for the corn event.

Claims

1. A pair of DNA molecules comprising:

a first DNA molecule and a second DNA molecule, wherein the DNA molecules are of sufficient length of contiguous nucleotides of SEQ ID NO;

3 or SEQ ID NO;

5 or the complement thereof to function as DNA primers or probes specific for DNA extracted from corn plant MON89034 or progeny thereof, wherein said first and second DNA molecules when used together in a DNA amplification method produce an amplicon comprising SEQ ID NO;

1 or the complement thereof.

2. The pair of DNA molecules of claim 1, wherein said first DNA molecule comprises 11 or more contiguous nucleotides of any portion of the transgene region of SEQ ID NO:

3 or SEQ ID NO;

5, or the complement thereof, and said second DNA molecule comprises 11 or more contiguous nucleotides of any portion of a 5′

flanking corn genomic DNA region of SEQ ID NO;

3, or the complement thereof.

3. The pair of DNA molecules of claim 2, wherein said first DNA molecule comprises at least 15 contiguous nucleotides fully complementary to the heterologous insert DNA portion of SEQ ID NO:

5, and wherein said second DNA molecule comprises at least 15 contiguous nucleotides from the corn genome portion of SEQ ID NO;

3.

4. The pair of DNA molecule of claim 2, wherein said first DNA molecule hybridizes specifically to SEQ ID NO:

5 from about nucleotide position 2060 through about nucleotide position 12,208, and wherein said second DNA molecule hybridizes specifically to the reverse complement sequence of SEQ ID NO;

3 from about nucleotide position 1 through about position 2050.

9. The pair of DNA molecules of any one of claims 1 to 8, wherein said first DNA molecule comprises SEQ ID NO:

6 and said second DNA molecule comprises SEQ ID NO;

7.

20. A DNA detection kit comprising a pair of DNA molecules according to any one of claims 1 to 9.

5. A pair of DNA molecules comprising:

a first DNA molecule and a second DNA molecule, wherein the DNA molecules are of sufficient length of contiguous nucleotides of SEQ ID NO;

4 or SEQ ID NO;

5 or the complement thereof to function as DNA primers or probes specific for DNA extracted from corn plant MON89034 or progeny thereof, wherein said first and second DNA molecules when used together in a DNA amplification method produce an amplicon comprising SEQ ID NO;

2 or the complement thereof.

6. The pair of DNA molecules of claim 5, wherein said first DNA molecule comprises 11 or more contiguous nucleotides of any portion of the transgene region of SEQ ID NO:

4 or SEQ ID NO;

5, or the complement thereof, and said second DNA molecule comprises 11 or more contiguous nucleotides of any portion of a 3′

flanking corn genomic DNA region of SEQ ID NO;

4, or the complement thereof.

7. The pair of DNA molecules of claim 6, wherein said first DNA molecule comprises at least 15 contiguous nucleotides from the heterologous insert DNA portion of SEQ ID NO:

5, and wherein said second DNA molecule comprises at least 15 contiguous nucleotides fully complementary to the corn genome portion of SEQ ID NO;

4.

8. The pair of DNA molecules of claim 6, wherein said first DNA molecule hybridizes specifically to the reverse complement sequence of SEQ ID NO:

5 from about nucleotide position 1 through about position 11305, and wherein said second DNA molecule hybridizes specifically to SEQ ID NO;

4 from about nucleotide position 21 through about nucleotide position 914.

10. A method of detecting the presence of a DNA molecule selected from the group consisting of SEQ ID NO:

3, SEQ ID NO;

4, and SEQ ID NO;

5 in a biological sample, the method comprising;

(a) contacting said biological sample with a DNA primer pair comprising DNA molecules of sufficient length of contiguous nucleotides of SEQ ID NO;

3 or the complement thereof, SEQ ID NO;

4 or the complement thereof, SEQ ID NO;

5 or the complement thereof, to function as DNA primers or probes specific for DNA extracted from corn plant MON89034 or progeny thereof, wherein said DNA primer pair when used together in a DNA amplification method produces an amplicon comprising SEQ ID NO;

1, SEQ ID NO;

2, or the complement thereof;

(b) providing a nucleic acid amplification reaction condition;

(c) performing said nucleic acid amplification reaction, thereby producing a DNA amplicon molecule; and

(d) detecting said DNA amplicon molecule, wherein detection of an amplicon comprising at least one of SEQ ID NO;

1, SEQ ID NO;

2 and the complement thereof is indicative of the presence of said DNA molecule in said biological sample.

11. The method of claim 10, wherein said biological sample is a DNA sample extracted from a corn plant.

16. The method of any one of claims 10 to 15, wherein said biological sample is selected from the group consisting of corn meal, corn oil, corn cake, corn seed, corn germ, corn starch, corn flour, corn pollen, corn silk, corn steep liquor, corn malt, corn sugar, corn syrup, margarine produced from corn oil, distillers dry goods solids (DDGS), cosmetic and bulking agent.

12. A method of detecting the presence of a DNA molecule selected from the group consisting of SEQ ID NO:

3, SEQ ID NO;

4 and SEQ ID NO;

5 in a biological sample, the method comprising;

(a) contacting said biological sample with a DNA probe that hybridizes under stringent conditions with said DNA molecule, and does not hybridize under the stringent conditions with a biological sample not containing said DNA molecule;

(b) subjecting said biological sample and DNA probe to stringent hybridization conditions; and

(c) detecting hybridization of said DNA probe to said biological sample, wherein detection of hybridization is indicative of the presence of said DNA molecule in said biological sample.

13. The method of claim 12, wherein said biological sample is a DNA sample extracted from a corn plant.

14. The method of claim 12, wherein said DNA probe comprises SEQ ID NO:

1 or SEQ ID NO;

2 or the complement thereof.

15. The method of claim 12, wherein said DNA probe is labeled with at least one fluorophore.

17. A DNA detection kit comprising:

at least one DNA molecule of sufficient length of contiguous nucleotides of SEQ ID NO;

3, SEQ ID NO;

4, SEQ ID NO;

5, or the complement thereof, to function as a DNA primer or probe specific for corn event MON89034 and/or its progeny.

18. The DNA detection kit of claim 17, wherein said at least one DNA molecule comprises SEQ ID NO:

1, SEQ ID NO;

2, or the complement thereof.

19. The DNA detection kit of claim 18, wherein said at least one DNA molecule is SEQ ID NO:

1, SEQ ID NO;

2, or the complement thereof.

21. A method of determining zygosity of DNA of a corn plant comprising corn event MON89034 in a biological sample comprising:

(a) contacting said sample with a primer set comprising SEQ ID NO;

6, SEQ ID NO;

7, and SEQ ID NO;

10, that (1) when used in a nucleic acid amplification reaction comprising corn event MON89034 DNA, produces a first amplicon that is diagnostic for corn event MON89034, and (2) when used in a nucleic acid amplification reaction comprising corn genomic DNA other than MON89034 DNA, produces a second amplicon that is diagnostic for corn genomic DNA other than MON89034 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 MON89034 DNA, wherein detection of only the first amplicon indicates that said sample is homozygous for corn event MON89034 DNA.

22. The method of claim 21, wherein said the primer set is further used together with SEQ ID NO:

14 and SEQ ID NO;

15.