中国HIV-1B'亚型流行株gp120蛋白的表达与纯化  

Expression and characterization of HIV-1 envelope glycoprotein gp120 derived from a Chinese B' isolate

作　　者：楚鹰[1] 宋红勇[1] 吴稚伟[1] 

机构地区：[1]南京大学医学院公共健康医学中心,江苏南京210093

出　　处：《东南大学学报（医学版）》2013年第6期742-745,共4页Journal of Southeast University(Medical Science Edition)

基　　金：国家自然科学基金面上项目(30870124);科技部国际合作项目(2009DFA31260)

摘　　要：目的:表达纯化中国HIV-1 B'亚型流行株gp120蛋白。方法:将gp120基因克隆至真核表达载体pTriEx-3 hygro,构建真核表达质粒pTriEx-3-gp120,转染CHO-K1细胞,96 h后收取上清,利用金属螯合层析的方法纯化该重组蛋白,并通过SDS-PAGE以及Western blotting验证重组蛋白的表达。结果:酶切及测序结果表明真核质粒表达构建正确,表达的重组蛋白可以被His标签抗体以及HIV-1阳性血清所识别,并从细胞转染上清中成功纯化了该蛋白。结论:我们成功构建了一个可用于免疫检测、疫苗设计以及其它相关研究的HIV-1免疫原。Objective：To express and character a recombinant HIV-1 gp120 （rgp120） derived from the env gene of a Chinese B＇subtype isolate .Methods：The gp120 gene was subcloned into pTriEx-3 hygro vector to construct eukaryotic expression plasmid pTriEx-3-gp120, which was transfected into CHO-K1 cell line.The expressed rgp 120 was purified by metal chelate affinity chromatography and characterized by SDS-PAGE and Western blotting . Results：The eukaryotic expression plasmid pTriEx-3-gp120 was successfully constructed and confirmed by enzyme digestion and sequencing .The rgp120 was identified by anti-his tag monoclonal antibody and HIV-1 positive sera , and can be isolated from cell culture supernatant by a simple and fast purification procedure based on metal chelate affinity chromatography .Conclusion：The protein may serve as a useful tool for studies relating to HIV-1 vaccine development , HIV-specific antibody responses and other relative research purposes .

关 键 词：HIV-1 GP120 蛋白表达纯化 

分 类 号：Q51[生物学—生物化学] Q786