Methods for generating polynucleotides having desired characteristics by iterative selection and recombination
AssigneeCodexis, Inc.
InventorsWillem P. C. Stemmer, Andreas Crameri
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Abstract
A method for DNA reassembly after random fragmentation, and its application to mutagenesis of nucleic acid sequences by in vitro or in vivo recombination is described. In particular, a method for the production of nucleic acid fragments or polynucleotides encoding mutant proteins is described. The present invention also relates to a method of repeated cycles of mutagenesis, shuffling and selection which allow for the directed molecular evolution in vitro or in vivo of proteins.
Claims6
A mutant green fluorescent protein (GFP) with enhanced fluorescent intensity as compared to wild type Aequorea victoria GFP, said mutant protein comprising point mutations at two or more residues corresponding to residues selected from the group consisting of F99, M153, and E172 in wild type A. victoria GFP, wherein said mutations comprise replacement of hydrophobic residues with more hydrophilic residues.
Dependencies are inferred from the claim text.