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Methods and compositions for polypeptide engineering

  • US 6,653,072 B1
  • Filed: 06/24/1999
  • Issued: 11/25/2003
  • Est. Priority Date: 12/18/1996
  • Status: Expired due to Fees
First Claim
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1. A method for improving expression of at least one protein, wherein the at least one protein is encoded by at least one DNA substrate molecule, comprising:

  • (a) providing a set of oligonucleotides, wherein each oligonucleotide comprises at least two regions complementary to the at least one DNA substrate molecule and at least one degenerate region, each degenerate region comprising a nucleic acid sequence that differs in at least one nucleotide from the complement of the at least one DNA substrate molecule;

    (b) assembling the set of oligonucleotides into a library of recombinant nucleic acids;

    (c) expressing the library of recombinant nucleic acids of (b) in a population of host cells; and

    (d) screening or selecting the population of host cells of (c) to identify at least one host cell expressing at least one protein encoded by at least one recombinant nucleic acid, wherein expression of the at least one protein encoded by the at least one recombinant nucleic acid in the at least one host cell is improved compared to expression of the at least one protein encoded by the at least one DNA substrate molecule in a host cell.

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