禽网状内皮组织增生病毒gp90蛋白的原核表达与纯化

Prokaryotic expression and purification of Reticuloendotheliosis virus gp90 protein

为了研究禽网状内皮组织增生病毒(REV)gp90蛋白的生物学功能和免疫学功能,合成了gp90蛋白基因,将该基因克隆至原核表达载体p ET-22b(+)上,转化至大肠杆菌BL21(DE3)感受态细胞,IPTG诱导剂诱导目的蛋白表达,并通过SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)和Western blotting鉴定了蛋白表达情况和蛋白表达形式。使用小规模蛋白纯化系统将表达的蛋白进行了纯化。结果表明:将gp90蛋白基因连接表达载体转化宿主菌后,成功在大肠杆菌上以包涵体形式表达,纯化的蛋白纯度超过90%。

In order to research the biological function and immunological function of gp90 protein of reticuloendotheliosis virus,the gp90 gene was synthesized and cloned into prokaryotic expression vector pET-22b( +). The recombinant vector was then transformed into E. coli BL21( DE3) competent cells and the gp90 protein was expressed by the inducing of IPTG. Furthermore,the expression and the expression form were identified by the method of SDS-polyacrylamide gel electrophoresis( SDS-PAGE) and Western blotting. Finally,the target protein was purified by protein purification mini Kit. The results showed that the gp90 gene was expressed in Escherichia coli successfully as inclusion body after being ligated with expression vector and transformed into host cells. The purity of the purified gp90 protein was more than 90%.