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Title: Method of generating ploynucleotides encoding enhanced folding variants

The invention provides directed evolution methods for improving the folding, solubility and stability (including thermostability) characteristics of polypeptides. In one aspect, the invention provides a method for generating folding and stability-enhanced variants of proteins, including but not limited to fluorescent proteins, chromophoric proteins and enzymes. In another aspect, the invention provides methods for generating thermostable variants of a target protein or polypeptide via an internal destabilization baiting strategy. Internally destabilization a protein of interest is achieved by inserting a heterologous, folding-destabilizing sequence (folding interference domain) within DNA encoding the protein of interest, evolving the protein sequences adjacent to the heterologous insertion to overcome the destabilization (using any number of mutagenesis methods), thereby creating a library of variants. The variants in the library are expressed, and those with enhanced folding characteristics selected.
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OSTI Identifier:
Los Alamos National Security, LLC LANL
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Resource Relation:
Patent File Date: 2008 Oct 02
Research Org:
Los Alamos National Lab. (LANL), Los Alamos, NM (United States)
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Country of Publication:
United States

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