Homologous recombination in plants
AssigneeMaxygen, Inc.
InventorsMichael Lassner, Steven delCardayre
Life of this patent
Estimating the term…
Abstract
Methods for evolving recombinase protein homologues and RecA/VirE2 fusion proteins which complement VirE2 deficient Agrobacterium are provided. The use of recombinase protein homologues and RecA/VirE2 fusion proteins in the context of Agrobacterium mediated transformation are provided. Methods for producing transgenic organisms by homologous recombination using evolved recombinase proteins and Agrobacterium strains which express recombinase protein homologues or RecA/VirE2 fusion proteins are provided. Transgenic cells and organisms which have integrated an exogenous DNA sequence into a predetermined site in their genome are provided.
Claims12
A method of evolving a recombinase to complement an Agrobacterium virE2 gene, the method comprising:
- (a) providing a population of nucleic acids, wherein the population encodes a plurality of recombinase genes;
- (b) recombining or mutating the population of nucleic acids to generate a library of recombinant nucleic acid segments;
- (c) optionally repeating the recombining or mutating process of steps (a) and (b) one or more times;
- and
- (d) screening the library of recombinant nucleic acid segments, which recombinant nucleic acid segments comprise evolved recombinase genes, to identify at least one recombinant nucleic acid segment encoding an evolved recombinase, which evolved recombinase can substitute functionally for an Agrobacterium virE2 gene in plant transformation.
Dependencies are inferred from the claim text.