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US 6,326,204 B1Granted
Evolution of whole cells and organisms by recursive sequence recombination
AssigneeMaxygen, Inc.
InventorsStephen delCardayre, Matthew Tobin, William P. C. Stemmer, Jon E. Ness, Jeremy Minshull, Phillip Patten, Venkiteswatan Subramanian, Linda Castle, Steve 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.
Claims49
A method of evolving a cell to acquire a desired function, comprising:
- (1) introducing a library of DNA fragments into a plurality of cells, whereby at least one of the fragments undergoes recombination with a segment in the genome or an episome of the cells to produce modified cells;
- (2) screening the modified cells for modified cells that have evolved toward acquisition of the desired function;
- (3) recombining DNA from the modified cells that have evolved toward the desired function with a further library of DNA fragments, at least one of which undergoes recombination with a segment in the genome or the episome of the modified cells to produce further modified cells, or recombining DNA between the modified cells that have evolved toward the desired function to produce further modified cells;
- (4) screening the further modified cells for further modified cells that have further evolved toward acquisition of the desired function;
- (5) repeating (3) and (4) as required until the further modified cells have acquired the desired function.
2 more elements
Dependencies are inferred from the claim text.