HIV跨膜蛋白gp36和gp41的截短及融合表达  

Truncation and Fusion Expression of Tans-membrane Proteins gp36 and gp41 of HIV

作　　者：肖健[1] 朱华松[1] 尹娟[1] 孙可芳[1] 汪超英[1] 端义坤[1] 余模松[1] 

机构地区：[1]武汉生物制品研究所,武汉430060

出　　处：《中国生物制品学杂志》2006年第2期124-126,138,共4页Chinese Journal of Biologicals

摘　　要：目的将HIV-1的跨膜蛋白gp41和HIV-2跨膜蛋白gp36进行截短,并在大肠杆菌中进行融合表达。方法用PCR将gp41和gp36的编码基因进行截短,回收的PCR产物纯化后克隆到连接载体pGEM-T上,然后用BamHⅠ、EcoRⅠ和SalⅠ切下目的基因,并构建到表达载体pGEX-4T-3,导入宿主细胞BL21,用IPTG诱导表达。结果酶切鉴定显示,截短的HIV-1gp41和HIV-2gp36跨膜蛋白基因大小与预期的一致,表达产物经SDS-PAGE分析显示在相对分子质量66000处出现融合表达条带,Westernblot分析显示,与相应抗体出现特异性反应。结论已成功对gp41和gp36跨膜蛋白进行截短,并构建表达载体进行表达,为跨膜蛋白的进一步应用研究奠定基础。Objective To truncate the trans-membrane proteins gp41 of HIV-1 and gp36 of HIV-2 and express in E. coli. Methods Truncate the genes encoding gp41 and gp36 by PCR,purify the PCR product and clone into vector pGEM-T. Digest the recombinant plasmid with BamH Ⅰ ,EcoR Ⅰ and Sal Ⅰ and insert the obtained target gene into expression vector pGEX-4T3. Transform the constructed recombinant plasmid to E. coli BL21 for expression under induction of IPTG. Identify the expressed product by SDSPAGE and Western blot. Results Restriction map proved that the lengths of truncated go41 and gp36 genes were identical to those expected. SDS-PAGE profile revealed a fusion expression band with a relative molecular weight of 66 000. Western blot showed specific reactions of expressed product with the corresponding antibodies. Conclusion The HIV-1 gp41 and HIV-2 gp36 genes were successfully truncated and expressed in E. coli. It laid a foundation of fur pplication of trans-membrane protein.

关 键 词：人类免疫缺陷病毒 跨膜蛋白 截短 融合表达 

分 类 号：R512.91[医药卫生—内科学] R392.11[医药卫生—临床医学]