不同区段HIV-1 env基因在Bac-to-Bac昆虫细胞杆状病毒表达系统中的表达及检测  

Expression and Detection of Different Fragments of HIV-1 Envelope Genes in a Bac-to-Bac Baculovirus Insect cell Expression System

作　　者：谢小燕[1] 郭庆[1] 程通[1] 李少伟[1] 张军[1] 夏宁邵[1] 

机构地区：[1]厦门大学细胞生物学与肿瘤细胞工程教育部重点实验室,厦门361005

出　　处：《病毒学报》2002年第1期15-20,共6页Chinese Journal of Virology

基　　金：国家重点科技 (攻关 )计划 ( 96 -92 0 -37-0 9) ;福建省计委重点攻关项目 ( 96 0 10 )

摘　　要：利用昆虫细胞杆状病毒表达系统 ,将从一株HIV - 1阳性克隆质粒中获得的几个HIV包膜蛋白基因片段 ,克隆入转移载体中得到重组病毒。用此重组病毒感染昆虫细胞后表达出 3种HIV包膜蛋白 ,即GP12 0 - 41P、GP41T、GP41P ,分别含有HIV - 1包膜糖蛋白GP12 0及部分GP41,删除了N端 12个疏水氨基酸的GP41和仅有主要表位约 2 40个氨基酸的GP41。收获后分别以Western -blotting和EIA检测 ,有较好的免疫学活性 ,其中GP41T的活性最强。该实验为HIV包膜蛋白的结构研究提供了依据 ,加以改进后可能有免疫检测的价值。Bac-to-Bac baculovirus expression vector system(BEVS)was used to express foreign proteins.Several HIV env genes were subcloned from a HIV-1 positive clone pNL4-3 to the transfer vector pFastBacl.The genes carried by pFastBacl were transported to baculovirus DNA-bacmid and formed recombinant bacmid within the DH10Bac E.coli strain.Then transfected and infected Sf9 insect cells with recombinant bacmid and baculovirus and obtained three HIV-1 env recombinant proteins:GP120-41P,GP41T and GP41P,representing HIV-1 envelope glycoprotein GP120 and part of GP41,trunscated GP41 with 12 hydrophobic amino acid deleted from GP41 5’-end and main epitope of GP41,respectively.The expressed products were harvested and analysed with Western-blot and EIA.They were found all had immune activity,and GP41T was the best one of all.Our work helps to study the structure of HIV envelope proteins.Once improved,it may be used in immunoassay.

关 键 词：Bac-to-Bac昆虫杆状病毒表达系统 人免疫缺陷病毒 包膜蛋白 抗原 表达 HIV-1 ENV基因 检测 

分 类 号：R373.9[医药卫生—病原生物学] R512.91[医药卫生—基础医学]