COMPOSITIONS AND METHODS FOR GENETIC MODIFICATION OF PLANTS

• US 20110030107A1
• Filed: 10/22/2010
• Published: 02/03/2011
• Est. Priority Date: 11/18/1997
• Status: Active Grant

• Current Assignee: Pioneer Hi Bred International Inc (Corteva, Inc.)
• Original Assignee: Pioneer Hi Bred International Inc (Corteva, Inc.)
• Inventors: Baszczynski, Christopher L., Bowen, Benjamin A., Tagliani, Laura A., Peterson, David J.
• Granted Patent: US 8,143,504 B2
• Time in Patent Office: Days
• US Class Current: 800/312
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C12N 15/8203 Virus mediated transformation
• CPC Class Codes: C12N 15/8205 Agrobacterium mediated transformation
• CPC Class Codes: C12N 15/8213 Targeted insertion of genes into the plant genome by homologous recombination
• CPC Class Codes: C12N 15/90 Stable introduction of foreign DNA into chromosome
• CPC Class Codes: C12N 9/00 Enzymes; Proenzymes; Compositions thereof preparations containing enzymes for cleaning teeth A61K8/66, A61Q11/00; medicinal preparations containing enzymes or proenzymes A61K38/43; enzyme containing detergent compositions C11D; enzymes with nucleic acid structure, e.g. ribozymes, C12N15/113; Processes for preparing, activating, inhibiting, separating or purifying enzymes preparation of malt C12C1/00

Abstract

Methods for the targeted integration of nucleotide sequences into a plant are provided. Transfer cassettes comprising nucleotide sequences of interest flanked by non-identical recombination sites are used to transform a plant comprising a target site. The target site contains at least a set of non-identical recombination sites corresponding to those on the transfer cassette. Exchange of the nucleotide sequences flanked by the recombination sites is effected by a recombinase.

Claims

1. A soybean plant cell having stably incorporated into its genome a transfer cassette comprising a nucleotide sequence of interest flanked by a first and a second non-identical recombination site, wherein said first and said second non-identical recombination site can recombine in said soybean plant cell with their identical recombination site in the presence of an appropriate recombinase.

2. The soybean plant cell of claim 1, wherein said first and said second non-identical recombination sites can recombine with their identical recombination sites in the presence of the appropriate recombinase when their identical recombination sites are sexually crossed into said soybean plant cell.

3. The soybean plant cell of claim 1, wherein said non-identical recombination sites comprise a FRT site, a mutant FRT site, a LOX site, or a mutant LOX site.

4. The soybean plant cell of claim 3, wherein said non-identical recombination sites comprises at least a FRT1 site (SEQ ID NO:

1), a FRT6 site (SEQ ID NO;

4), a FRT7 site (SEQ ID NO;

5) or a FRT5 site (SEQ ID NO;

3).

5. The soybean plant cell of claim 1, wherein the genome further comprises an expression cassette comprising a nucleotide sequence encoding the recombinase.

6. The soybean plant cell of claim 4, wherein said recombinase comprises a FLP recombinase.

7. The soybean plant cell of claim 3, wherein said recombinase comprises a Cre recombinase.

8. The soybean plant cell of claim 1, wherein said soybean plant cell has stably incorporated into its genome a polynucleotide comprising in the following order, at least one expression cassette comprising a second nucleotide sequence of interest and the transfer cassette.

9. The soybean plant cell of claim 1, wherein said plant cell has stably incorporated into its genome a polynucleotide comprising in the following order, at least one expression cassette comprising a second nucleotide sequence of interest, a promoter, the transfer cassette comprising the nucleotide sequence of interest operably linked to said promoter.

10. The soybean plant cell of claim 1, wherein said nucleotide sequences of interest is selected from the group consisting of a sequence that influences plant growth, the susceptibly of the plant to disease, the susceptibly of the plant to insects, and the nutritional value of the plant.

11. A soybean plant having stably incorporated into its genome a transfer cassette comprising a nucleotide sequence of interest flanked by a first and a second non-identical recombination site, wherein said first and said second non-identical recombination site can recombine in said soybean plant with their identical recombination site in the presence of an appropriate recombinase.

12. The soybean plant of claim 11, wherein said first and said second non-identical recombination sites can recombine with their identical recombination sites in the presence of the appropriate recombinase when their identical recombination sites are sexually crossed into said soybean plant.

13. The soybean plant of claim 11, wherein said non-identical recombination sites comprise a FRT site, a mutant FRT site, a LOX site, or a mutant LOX site.

14. The soybean plant of claim 13, wherein said non-identical recombination sites comprises at least a FRT1 site (SEQ ID NO:

1), a FRT6 site (SEQ ID NO;

4), a FRT7 site (SEQ ID NO;

5) or a FRT5 site (SEQ ID NO;

3).

15. The soybean plant of claim 11, wherein the genome further comprises an expression cassette comprising a nucleotide sequence encoding the recombinase.

16. The soybean plant of claim 15, wherein said recombinase comprises a FLP recombinase.

17. The soybean plant of claim 15, wherein said recombinase comprises Cre recombinase.

18. The transformed seed of the plant of claim 11.

19. The soybean plant of claim 11, wherein said soybean plant has stably incorporated into its genome a polynucleotide comprising in the following order, at least one expression cassette comprising a second nucleotide sequence of interest and the transfer cassette.

20. The soybean plant of claim 11, wherein said soybean plant has stably incorporated into its genome a polynucleotide comprising in the following order, at least one expression cassette comprising a second nucleotide sequence of interest, a promoter, the transfer cassette comprising the nucleotide sequence of interest operably linked to said promoter.