IN-VIVO CLONING OF PCR PRODUCTS IN ESCHERICHIA-COLI

This report describes an efficient method to clone PCR products exploi ting endogenous Escherichia coli enzymatic activities. PCR products ar e engineered to contain terminal sequences identical to sequences at t he two ends of a linearized vector. PCR products and vector DNA are th en simply co-transfected into E. coli strain JC8679, obviating the req uirement for enzymatic treatment of the PCR product or in vitro ligati on. The high rate of homologous recombination in this strain results i n efficient incorporation of the insert into the vector, a process we refer to as in vivo cloning (IVC).