大肠杆菌素通道结构域与变异链球菌感受刺激多肽原核载体的构建和表达

Construction,expression,and purification of colicin Ia channel domain-CSP fusion protein

目的:表达并纯化大肠杆菌素Ia通道结构域(Colicin Ia channel domain)与变异链球菌感受刺激多肽(CSP)的融合蛋白。方法:将编码大肠杆菌素Ia通道结构域的基因与编码变异链球菌感受刺激多肽CSP的基因导入带有麦芽糖结合蛋白(MBP)基因的pMAL-c2x质粒中,构建pMAL-c2x-Colicin Ia channeldomain-CSP融合表达载体,将重组质粒转化大肠埃希菌BL21后,IPTG诱导融合蛋白表达,裂解后得到目的蛋白。结果:通过双酶切、电泳分析、测序,证明成功构建了pMAL-c2x-Colicin Ia channel domain-CSP融合表达载体,目的蛋白产物经SDS-PAGE分析,与预期一致。结论:成功构建了pMAL-c2x-Colicin Ia channeldomain-CSP,表达并纯化目的蛋白,为进一步研究Colicin Ia channel domain-CSP的功能奠定了基础。

AIM： To express and purify the colicin Ia channel domain -CSP fusion protein. METHODS： The fragments of Coliein Ia channel domain and CSP were inserted into pMAL - c2x containing maltose - binding protein （MBP） to construct pMAL - c2x - Colicin la channel domain - CSP fusion expression vector. The plasmid was transformed into E. coli BL21 and induced by IPTG for fusion protein which was digested to obtain the aimed protein. RESULTS： Through double - digestion, electrophoresis and sequencing, the successful construction of the pMAL - c2x - Coliein Ia channel domain - CSP fusion expression vector was proved. After SDS - PAGE analysis the aimed protein was consistent with expected. CONCLUSION ： The pMAL - e2x - Coliein la channel domain - CSP was construeted successfully. The aimed protein was expressed and purified, which lay a basis for future research.