热休克蛋白DnaK在大肠杆菌中的克隆与表达

Cloning and Expression of Heat Shock Protein DnaK in Escherichia coli

PCR扩增了热休克蛋白DnaK(HSP70)基因,将其连接到pBAD Directional TOPO载体上,构建了重组表达质粒,转化至One ShotTOP 10中的大肠杆菌Top 10中,经阿拉伯糖诱导获得重组蛋白。通过Western blot验证,表明该目标蛋白具有与鼠抗DnaK单抗特异结合的抗原活性,说明表达的重组蛋白为Hsp70,该研究为进一步探讨Hsp70的生物学功能和作用机制奠定了基础。

DnaK（Hsp70）was amplified by PCR and cloned into TOPO cloning vector using a pBAD Directional TOPO Expression Kit.With L-arobinose induction,DnaK（Hsp70）has been successfully expressed in Top 10 E.coli cells.Western-blot showed the specific reaction of the expressed protein with monoclonal mouse anti-DnaK,thus confirmed the recombinant protein was Hsp70,which laid a foundation for further study on its biological functions and action mechanisms.