Institute of Molecular Biology, Biology Faculty of Hubei University, Wuhan, Hubei Province, 430062, People's Republic of China.
Biotechnol Lett. 2010 Mar;32(3):439-43. doi: 10.1007/s10529-009-0166-3.
Novel directional cloning and expression vectors were developed for blunt-end ligation of PCR products that are suitable for high-throughput cloning and simplifying the screening procedure. The PCR products, without further processing, are cloned into vectors digested with SchI and, following transformation, the desired recombinants give typical blue colonies on selectable plates. The principle of this selection strategy is that the construction also generates a full-length ideal lacO gene. To the best of our knowledge, this is the first time that this lacO reconstruction strategy has been applied in the selection of recombinants.
新型定向克隆和表达载体已被开发用于 PCR 产物的平端连接，非常适合高通量克隆和简化筛选过程。无需进一步处理，PCR 产物即可被克隆到用 SchI 消化的载体中，转化后，所需的重组体在选择性平板上产生典型的蓝色菌落。这种选择策略的原理是构建物还生成全长理想的 lacO 基因。据我们所知，这是首次将这种 lacO 重建策略应用于重组体的选择。
