播娘蒿DsCOR基因的原核表达、蛋白纯化、及性质研究

A study on the protein expressed,purification,and character of DsCOR Gene from Descurania sophia(L.) Webb

利用PCR方法扩增DsCOR基因的蛋白编码序列,经BamHⅠ、SacⅠ酶切后连接入pET32a原核表达载体,构建的pET32a-DsCOR融合重组表达质粒转化大肠杆菌BL21进行原核表达.建立了"煮沸-镍离子亲和层析"的蛋白纯化方法.对纯化的DsCOR蛋白进行高温耐受性、pH耐受范围、紫外耐受性方面的研究.

The ORF of DsCOR was amplified by PCR, and cloned into expression vector pET32a to construct recombinant expression plasmid pET32a-DsCOR. The recombinant plasmid was transformed into E. coli BL21 and induced to express recombinant protein with IPTG. According to hydrophilic character, The fusion protein was further purified through boiling and affinity chromatography; concentrated by PEG; analyzed by SDS-PAGE. Research about protein resistance to heat, acid, alkali and UV was done.