HIV-2env(gp120)基因在大肠杆菌中的表达  

Expression of HIV-2 env (gp120) gene in E.coli

作　　者：王秀清[1] 金宁一[1] 田梅[1] 毛春生[1] 郭军庆[1] 顾万钧[1] 佟明华[1] 殷震[1] 

机构地区：[1]解放军农牧大学军事兽医研究所 [2]白求恩医科大学应用基础医学研究所 [3]白求恩医科大学预防医学院

出　　处：《白求恩医科大学学报》1997年第6期591-593,共3页Journal of Norman Bethune University of Medical Science

摘　　要：ＨＩＶ－２Ｅｎｖ蛋白（ｇｐ１２０）是病毒粒子吸附靶细胞膜上ＣＤ４受体的关键蛋白，ｇｐ１２０与ＣＤ４受体结合过程是ＨＩＶ感染机体的第一步。本文应用ＤＮＡ重组技术将ＨＩＶ－２ｅｎｖ（ｇｐ１２０）基因克隆到ｐＥＴ１７ｂ的Ｔ７启动子下游ＥｃｏＲＶ位点，构建成ｐＥＴ１７ｂ－ｇｐ１２０重组质粒，在大肠杆菌ＢＬ２１（ＤＥ３）中表达。ＳＤＳ－ＰＡＧＥ结果显示在３５０００（Ｍｒ）处有一特异蛋白带，表达量占菌体总蛋白的５．８％。蛋白印迹试验分析此蛋白能与ＨＩＶ－２阳性血清发生特异性反应。ＨＩＶ－２ｅｎｖ基因在大肠杆菌中的表达成功，为ＨＩＶ诊断试剂和疫苗的研究提供有益资料。The cDNA fragment of HIV-2 env gp120 (Human Immunodeficiency Virus Type 2 envelope gp120) gene was obtained after the plasmid pCRTMⅡ containing HIV-2 gp120 gene was degraded with BgIⅡ.The fragment was subcloned into pET17b-EcoR V restriction site under the control of bacterophage T7 promoter.The recombiniant plasmid pET17b-gp120 was constructed and expressed in E.coli BL21(DE3).SDS-PAGE (Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis) results proved that HIV-2 gp120 showed a single band with a molecular weight of 35 000 Dalton,and the expressive level was 5.8% of the total bacterial protein.Western blot demonstrated that the HIV-2 gp120 showed a specific reaction with the seria of HIV-2 patients.

关 键 词：艾滋病素-2 大肠杆菌 基因表达 

分 类 号：R373.9[医药卫生—病原生物学] R378.2[医药卫生—基础医学]