Methods and compositions for polypeptide engineering
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Abstract
Methods are provided for the evolution of proteins of industrial and pharmaceutical interest, including methods for effecting recombination and selection. Compositions produced by these methods are also disclosed.
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Citations
284 Claims
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1-273. -273. (canceled)
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274. A method for producing and isolating a polypeptide having at least one desirable property comprising:
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subjecting a starting polynucleotide set to a mutagenesis process so as to produce a set of progeny polynucleotides;
subjecting said progeny polynucleotide set to an end selection-based screening and enrichment process so as to select for a desirable subset of the progeny polynucleotide set, said end-selection process resulting in ligation compatible ends that can be used to facilitate one or more intermolecular ligations by members of the progeny polynucleotide set;
producing a polypeptide set by expression cloning, comprising ligating members of said progeny polynucleotide set into an expression vector using said ligation compatible ends; and
screening said polypeptide set for said at least one desirable property to identify polypeptides comprising said desirable property. - View Dependent Claims (275, 276, 277, 278, 279, 280, 281, 282, 283, 284)
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Specification