HIV-1CRF07B/C亚型gp41的表达、纯化和初步结构分析  

Prokaryotic Expression,Purification and Crystallization of HIV-1 CRF07 B/C Subtype gp41 Helix-bundle

作　　者：王建新[1] 王敏[1] 张同存[1] 刘新奇[2] 

机构地区：[1]天津科技大学生物工程学院,天津300457 [2]南开大学生命科学学院,天津300071

出　　处：《生物技术进展》2011年第1期56-60,F0003,共6页Current Biotechnology

基　　金："艾滋病和病毒性肝炎等重大传染病防治"专项(2008ZX10001-010);国家973计划项目(2010CB911800)资助

摘　　要：HIV-1跨膜蛋白gp41是HIV-1包膜与靶细胞膜融合过程中的关键蛋白,而且序列保守,是理想的HIV-1作用靶点。为获得HIV-1中国流行株CRF07B/Cgp41蛋白的晶体结构来指导疫苗设计及药物开发,采用CRF07B/Cgp160基因序列为模板,经PCR、酶切、连接,将gp41helix-bundle区域克隆到pET30-his表达载体中,经表达、纯化和结晶筛选,获得了gp41 helix-bundle的晶体并解析了结构,为针对中国艾滋病病毒流行株疫苗的设计及药物开发提供了结构参考。HIV-1 transmembrane protein gp41, which plays an important role in the fusion of viral and cellular membranes, is the ideal target for intervention of HIV-1 infection. In order to obtain gp41 crystal structure from prevalent strain in China, gp41 helix-bundle region was cloned into pET30-his vector by overlap PCR, enzyme digestion and ligation using CRF07 B/C gp160 gene as template. The plasmid was transferred into E. coli Rosetta （DE3）, and expression, purification and crystal screening of gp41 were conducted. Crystals of gp41 helix-bundle were obtained and its structure was solved by molecular replacement method. The overall structure was similar to B subtype, but the unique characteristics in local conformation would facilitate the vaccine design and drug development against CRF07 B/C prevalent in China.

关 键 词：HIV-1 GP41 表达 纯化 晶体结构 

分 类 号：R512.91[医药卫生—内科学]