葡萄球菌蛋白A抗体结合区基因的克隆、表达和纯化

目的构建表达金黄色葡萄球菌A蛋白(SPA)抗体结合区基因SPA-ZZ的pET-32a-ZZ表达载体,在大肠杆菌BL21中进行高效表达并初步纯化。方法用PCR从pEZZ18中克隆SPA抗体结合区基因ZZ,构建重组表达质粒pET32-ZZ,经酶切和测序确认,将阳性重组质粒转化BL21感受态细菌,IPTG诱导表达,表达产物进行聚丙烯酰胺凝胶电泳检测(SDS-PAGE),并用Q-Sephrose Fast Flow柱进行初步纯化。结果得到了高效表达SPA-ZZ的pET32-ZZ表达载体,表达产物经SDS-PAGE检测相对分子质量为41 000,初步纯化后可得到较纯的表达蛋白。结论SPA抗体结合区蛋白ZZ能在大肠杆菌中高效表达,为SPA-ZZ的应用奠定了一定的基础。

Cloning,expression and purification of Staphylococcal Protein A antibody-binding fraction gene

Purpose To construct an E.coli expression system pET32a-ZZ of Staphylococcal Protein A(SPA) antibody-binding fraction ZZ gene,to over-express and to purify the protein ZZ.Methods SPA-ZZ gene was amplified with polymerase chain reaction(PCR) from pEZZ18,and inserted into the expression vector pET32a,and the recombinant plasmid pET32a-ZZ was transformed into E.coli BL21,and induced by IPTG.The expressed protein was examinated by SDS-PAGE,and purified in Q-Sephrose Fast Flow.Results Both cloning and expression...