NOVEL DNA CLONING METHOD RELYING ON THE E. COLI RECE/RECT RECOMBINATION SYSTEM

摘要

The invention refers to a novel method for cloning DNA molecules using a homologous recombination mechanism between at least two DNA molecules comprising: a) providing a host cell capable of performing homologous recombination, b) contacting in said host cell a first DNA molecule which is capable of being replicated in said host cell with a second DNA molecule comprising at least two regions of sequence homology to regions on the first DNA molecule, under conditions which favour homologous recombination between said first and second DNA molecules and c) selecting a host cell in which homologous recombination between said first and second DNA molecules has occurred. In particular, it relies on the use of the E. coli RecE and RecT proteins, the bacteriophage Red-alpha and Red-beta proteins, or the phage P22 recombination system. The beneficial effects of concomitant expression of the RecBC inhibitor genes (e.g. Red-Gamma) is also examplified.