DNA克隆载体的"三片段克隆法"改造策略

A General Method for Direct DNA Vector Modification

Cloning one or more DNA fragments into a vector is the one of most commonly used techniques in molecular biology. Classic cloning strategy, when dealt with complicated cloning steps, like multiple fragments insertion, selective marker exchange, are often troubled by the choice of restriction enzymes, the base mutations caused by long fragment PCR, UV exposure and gel purification. We made use of the Red/ET technology to clone two fragments into the vector while remove the unnecessary vector part in a single step. This three pieces cloning manipulation is straightforward and convenient, with the potential to be a general cloning strategy for vector engineering.