HIVⅠ型外膜蛋白/干扰素A-2b的构建与表达  

Construction and expression of recombinant plasmid and expression of fusion of HIV-1 env/IFNα-2b

作　　者：关国坤[1] 冯立[2] 王洪军[2] 张志强[2] 王立强[2] 吴益民[2] 

机构地区：[1]中国人民解放军第211医院,哈尔滨150080 [2]沈阳军区疾病预防控制中心

出　　处：《中国公共卫生》2006年第7期835-836,共2页Chinese Journal of Public Health

基　　金：国家自然科学基金项目(3977066)

摘　　要：目的构建艾滋病病毒外膜蛋白(env)与干扰素(IFNα-2b)融合基因表达质粒,观察其在痘苗病毒中共表达结果。方法利用基因重组技术,将IFNα-2b基因片段插入到env基因的nt8383位点,经脂质体转染与血凝素阴性蚀斑筛选,挑出重组痘苗病毒。经免疫荧光、免疫印迹(Western blot)和斑点酶联免疫吸附试验(Dot-ELISA)鉴定表达产物。结果间接免疫荧光实验结果显示,转染重组质粒的细胞表面有绿色荧光。Western-blot与Dot-ELISA结果均显示,重组质粒转染细胞的裂解物中存在表达的env/IFNα-2b蛋白。结论成功地构建了重组真核细胞表达质粒,表达的蛋白具有良好的免疫原性与免疫反应性。Objective To construct an eukaryotic expression plasmid and express fusion of HIV - 1 env/IFNα - 2b in BHK21 cell. Methods An eukaryotic expression plasmid pSFJ16env/IFNα - 2b gene into vaccine vector plasmid pSFJ16 and transfected to BHK21 cell in vitro by liposome - mediated method. The expressed product was detected by indirect immunfluorescence assay（IFA）, western blot and Dot - ELISA. Results Indirect IFA showed green fluorescence on the surface of transfected cell. Both western blot and Dot- ELISA proved that the lysate of transfected cells contained env/IFNα- 2b protein.Conelusion An eukaryotic expression plasmid for HIV - 1 env/IFNα- 2b was successfully construced, and the expressed product showed good reactinogenicity and immunogenieity.

关 键 词：痘苗病毒 免疫原 人类免疫缺陷病毒 

分 类 号：R512.91[医药卫生—内科学]