Evolution of whole cells and organisms by recursive sequence recombination
AssigneeMaxygen, Inc.
InventorsStephen delCardayre, Matthew Tobin, Willem P. C. Stemmer, Jon E. Ness, Jeremy Minshull, Phillip Patten, Venkiteswaran Subramanian, Linda Castle, Claus M. Krebber, Steven H. Bass
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Abstract
The invention provides methods employing iterative cycles of recombination and selection/screening for evolution of whole cells and organisms toward acquisition of desired properties. Examples of such properties include enhanced recombinogenicity, genome copy number, and capacity for expression and/or secretion of proteins and secondary metabolites.
Claims48
A method of evolving a cell toward acquisition of a desired property, comprising:
- (a) introducing a DNA fragment library cloned into an artificial chromosome into a population of cells;
- (b) culturing the cells under conditions whereby sexual recombination occurs between the cells, whereby DNA fragments cloned into the artificial chromosome homologously recombine with corresponding segments of endogenous chromosomes of the population of cells, and endogenous chromosomes recombine with each other;
- (c) screening or selecting for cells that have evolved toward acquisition of the desired property.
A method of evolving a DNA segment for acquisition of a desired property, comprising:
- (a) providing a library of variants of the segment, each variant cloned into separate copies of an artificial chromosome;
- (b) introducing the copies of the artificial chromosome into a population of cells;
- (c) culturing the cells under conditions whereby sexual recombination occurs between cells and homologous recombination occurs between copies of the artificial chromosome bearing the variants;
- (d) screening or selecting for variants that have evolved toward acquisition of the desired property.
Dependencies are inferred from the claim text.