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METHODS AND COMPOSITIONS FOR CELLULAR AND METABOLIC ENGINEERING

  • US 20100144547A1
  • Filed: 09/11/2009
  • Published: 06/10/2010
  • Est. Priority Date: 02/17/1994
  • Status: Active Grant
First Claim
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1. A method of evolving a biocatalytic activity of a cell, comprising:

  • (a) recombining at least a first and second DNA segment from at least one gene conferring ability to catalyze a reaction of interest, the segments differing from each other in at least two nucleotides, to produce a library of recombinant genes;

    (b) screening at least one recombinant gene from the library that confers enhanced ability to catalyze the reaction of interest by the cell relative to a wildtype form of the gene;

    (c) recombining at least a segment from the at least one recombinant gene with a further DNA segment from the at least one gene, the same or different from the first and second segments, to produce a further library of recombinant genes;

    (d) screening at least one further recombinant gene from the further library of recombinant genes that confers enhanced ability to catalyze the reaction of interest by the cell relative to a previous recombinant gene;

    (e) repeating (c) and (d), as necessary, until the further recombinant gene confers a desired level of enhanced ability to catalyze the reaction of interest by the cell.

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