Method and compositions for directed clonning and subclonning using homologous recombination
A technology of homology arm and recombinase, applied in the field of cloning
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[0058] The present invention relates to a method and a composition for DNA cloning and subcloning using bacterial recombinase-mediated homologous recombination technology. The inventors of the present invention have found that bacterial recombinases can be used in a specific manner to obtain highly efficient target cloning or subcloning.
[0059]Bacterial recombinases preferably use RecE / T and / or Redα / β. In E. coli, the RecE / T pathway has been previously described and its components partially recognized (Hall and Kolodner, 1994, Proc. Natl. Acad. Sci. U.S.A. 91: 3205-3209; Gillen et al., 1981, J. Bacteriol. 145:521-532). Recombination through the RecE / T pathway requires the expression of two genes, rec E and rec T, the DNA sequences of which have been published (Hall et al., 1993, J. Bacteriol. 175:277-278). The function of the Rec E protein is similar to that of the λ exon, which is also called Redα, and the function of the Rec T protein is similar to that of the Redβ and e...
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