Compositions and methods for genetic modification of plants

• US 8,536,420 B2
• Filed: 08/01/2008
• Issued: 09/17/2013
• Est. Priority Date: 11/18/1997
• Status: Expired due to Fees

• Current Assignee: Pioneer Hi-Bred International (Corteva, Inc.)
• Original Assignee: Pioneer Hi-Bred International (Corteva, Inc.)
• Inventors: Baszczynski, Christopher L., Bowen, Benjamin A., Peterson, David J., Tagliani, Laura A.
• Primary Examiner(s): Zheng, Li
• Application Number:
  US12/184,564
• Publication Number:
  US 20080320617A1
• Time in Patent Office: 1,873 Days
• Field of Search: None
• US Class Current: 800/320.1
• 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 maize 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 FRT recombination site, wherein said first and said second non-identical recombination site can recombine in said maize plant cell with their identical recombination site in the presence of the same recombinase.

2. The maize plant cell of claim 1, wherein said non-identical FRT recombination sites comprise an FRT site or a mutant FRT site.

3. The maize plant of claim 1, wherein the genome further comprises an expression cassette comprising a nucleotide sequence encoding a FLP recombinase.

4. The maize plant cell of claim 3, wherein said nucleotide sequence encoding the FLP recombinase comprises maize preferred codons.

10. The maize plant cell of claim 1, wherein said maize 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.

11. The maize 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.

14. The 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.

5. A maize 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 FRT recombination site, wherein said first and said second non-identical FRT recombination site can recombine in said maize plant with their identical recombination site in the presence of the same recombinase.

6. The maize plant of claim 5, wherein said non-identical recombination sites comprise a FRT site or a mutant FRT site.

7. The maize plant of claim 5, wherein the genome further comprises an expression cassette comprising a nucleotide sequence encoding a FLP recombinase.

8. The maize plant cell of claim 7, wherein said nucleotide sequence encoding the FLP recombinase comprises maize preferred codons.

12. The maize plant of claim 5, wherein said maize 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.

13. The maize plant cell of claim 5, wherein said maize 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.

9. A transformed maize seed having stably incorporated into its genome a transfer cassette comprising a nucleotide sequence of interest flanked by a first and a second non-identical FRT recombination site, wherein said first and said second non-identical recombination site can recombine in a maize plant with their identical recombination site in the presence of the same recombinase.