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Methods for making character strings, polynucleotides and polypeptides having desired characteristics

  • US 7,058,515 B1
  • Filed: 03/30/2000
  • Issued: 06/06/2006
  • Est. Priority Date: 01/19/1999
  • Status: Expired due to Term
First Claim
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1. A method of making a recombinant nucleic acid, the method comprising:

  • providing a plurality of parental character strings corresponding to a plurality of nucleic acids, which character strings, when aligned for maximum identity, comprise at least one region of heterology, whereby each parental character string when so aligned contains areas of identity and areas of heterology with any other parental character string;

    aligning the character strings;

    defining a set of character string subsequences, which set of subsequences comprises subsequences of at least two of the plurality of parental character strings;

    providing a set of oligonucleotides corresponding to the set of character string subsequences;

    annealing members of the set of oligonucleotides with one another; and

    ,elongating one or more members of the set of oligonucleotides with a polymerase, or ligating at least two members of the set of oligonucleotides with a ligase, thereby producing one or more recombinant nucleic acids,wherein the plurality of parental character strings comprises at least two parental character strings,wherein the oligonucleotide set comprises at least one oligonucleotide member comprising a chimeric nucleic acid sequence that comprises a subsequence from each of at least two parental character strings,wherein the subsequences from each of the at least two parental character strings in the chimeric nucleic acid are separated by a crossover point, andwherein at least one crossover point for at least one oligonucleotide member is selected from a region outside of an identified pairwise homology region.

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