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US 6,251,674 B1Granted
Evolution of whole cells and organisms by recursive sequence recombination
AssigneeMaxygen, Inc.
InventorsMatthew Tobin, William P. C. Stemmer, Jon E. Ness, Jeremy Minshull
Life of this patent
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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.
Claims5
A method of predicting efficacy of a drug in treating a viral infection, comprising
- (1) recombining a nucleic acid segment from a virus, whose infection is inhibited by a drug, with at least a second nucleic acid segment from the virus, the second nucleic acid segment differing from the nucleic acid segment in at least two nucleotides, to produce a library of recombinant nucleic acid segments;
- (2) contacting host cells with a collection of viruses having genomes including the recombinant nucleic acid segments in a media containing the drug, and collecting progeny viruses resulting from infection of the host cells,
- (3) recombining a recombinant DNA segment from a first progeny virus with at least a recombinant DNA segment from a second progeny virus to produce a further library of recombinant nucleic acid segments;
- (4) contacting host cells with a collection of viruses having genomes including the further library or recombinant nucleic acid segments, in media containing the drug, and collecting further progeny viruses produced by the host cells,
- (5) repeating (3) and (4), as necessary, until a further surviving organism has acquired a degree of resistance to the drug, whereby the degree of resistance acquired, the number of repetitions of (3) and (4), or the number of mutations needed to acquire the degree of resistance are used to provide a measure of the ability of the drug to avoid field resistance.
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A method of predicting efficacy of a drug in treating an infection by a pathogenic microorganism, comprising
- (1) transforming a plurality of cells of the microorganism with a library of DNA fragments at least some of which undergo recombination with segments in the genome of the cells to produce modified microorganism cells;
- (2) propagating modified microorganisms in a media containing the drug, and recovering surviving microorganisms;
- (3) recombining DNA from surviving microorganisms with a further library of DNA fragments at least some of which undergo recombination with cognate segments in the DNA from the surviving microorganisms to produce further modified microorganisms cells;
- (4) propagating further modified microorganisms, in media containing the drug, and collecting further surviving microorganisms;
- (5) repeating (3) and (4), as necessary, until a further surviving microorganism has acquired a degree of resistance to the drug, whereby the degree of resistance acquired, the number of repetitions of (3) and (4), or the number of mutations needed to acquire the degree of resistance are used to provide a measure of the ability of the drug to avoid field resistance.
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Dependencies are inferred from the claim text.