HIV-1外膜蛋白原核表达载体的构建与表达

为原核表达免疫缺陷病毒(human immunodeficiency virus,HIV)外膜蛋白,对HIV-1外膜蛋白的基因进行了修饰,并利用PCR技术克隆env基因,将env基因克隆到原核表达载体中,利用大肠杆菌表达系统表达外膜蛋白,应用Western blotting检测其表达情况.结果表明:酶切鉴定证实正确地构建了pRSETB表达质粒,Western blotting和SDS-PAGE试验检测结果表明,构建后的env基因能在低温诱导的条件下表达.产物的相对分子质量为50000和33000,表达的env蛋白具有较好的免疫原性.

Construction and expression of prokaryotic expression vector of HIV-1 membrane protein

To construct prokaryotic expressing vector of HIV-1 membrane protein,the HIV-1 membrane protein genes was decorated,the env genes were obtained by PCR technology,they were cloned into the prokaryotic expression vector,and the coli expression system was applied to express the membrane protein,products were examined by Western blotting.Results shows that:the expression plasmid of pRSET B was contructed successfully confirmed by enzyme digestion identification,the Western blot and SDS-PAGE analysis show that env protein can be expressed in E.coli BL21 at low temperature.Relative molecular mass of the products were 50 000 and 33 000,proteins shows a good reactionogenicity as a result.