表达中国流行株HIV-1 gp120外膜蛋白靶细胞模型的建立  

Establishment of Targets Cells Expressing Surface Glycoprotein gp120 of Chinese HIV-1 Strain

作　　者：李昌[1] 金宁一[2] 胡宁宁[2] 刘玉生[2] 佟敬山[2] 李霄[2] 金洪涛[2] 于芳[2] 

机构地区：[1]吉林大学畜牧兽医学院,长春130062 [2]军事医学科学院军事兽医研究所,长春130062

出　　处：《中国生物工程杂志》2008年第4期12-16,共5页China Biotechnology

基　　金：吉林省科技厅项目(20060570);国家“863”计划(2006AA02Z447);长春市科技局项目(2007KZ19)资助项目

摘　　要：利用pDispaly真核表达载体构建了用于表达中国流行株HIV-1gp120的真核表达质粒pD-120,通过脂质体转染法将其导入人宫颈癌细胞(HeLa),经G418反复加压筛选,直到细胞不再死亡为止,8代后获得稳定表达中国流行株HIV-1gp120蛋白的靶细胞复制模型HeLa-gp,SDS-PAGE,蛋白印迹与间接免疫荧光分析表明,重组蛋白得到很好表达,并具有良好生物活性,为抗AIDS/HIV治疗用基因工程制剂或靶向药物的活性检测奠定了坚实基础。Recombinant eukaryotic expression plasmid pD-120 containing gpl20 gene of Chinese HIV-1 strain was constructed successfully by using pDisplay vector. After transfected the plasmid into HeLa cells with Lipofectamine transfection method, the HeLa-gpl20 transfectants were obtained after 8 generations by pressure screening with G418 until no death of cells occur. The expressed products were analyzed by SDS-PAGE, Western blot and indirect immunofluorescent assay. The result shows that target protein was successfully expressed and the biology-activities were very well. This will settle solid foundation for biology-activity detection of genetic agents or target medicine of AIDS/HIV treatment.

关 键 词：HIV.1 GP120 HeLa细胞模型建立 

分 类 号：Q813[生物学—生物工程]