Patent ID: 6358742
Filing Date: 2002-03-19
Classification: C12N

Abstract:
A method of evolving a gene to confer enhanced conjugative transfer, comprising:(1) recombining at least first and second DNA segments from at least one conjugative transfer gene, the segments differing from each other in at least two nucleotides, to produce a library of recombinant genes; (2) screening the library for at least one recombinant gene from the library that confers enhanced conjugal transfer between cells relative to a wild-type form of the gene; (3) recombining at least a segment from the at least one recombinant gene with a further DNA segment from the at least one conjugative transfer gene, the same or different from the first and second segments, to produce a further library of recombinant genes; (4) screening the further library for at least one further recombinant gene from the further library of recombinant genes that confers enhanced conjugal transfer between cells relative to the at least one recombinant gene; and (5) repeating (3) and (4), as necessary, until the further recombinant gene confers a desired level of enhanced conjugal transfer between cells.