HIV-1 gag/IFNα-2b表达产物的免疫原性研究  

Study on Immunogenicity of HIV-1 gag/ IFNα-2b Expression Product

作　　者：王洪军[1] 孙艳[2] 周青阳[1] 刘宏波[1] 许志伟[1] 吴扬[3] 孙淑艳[3] 

机构地区：[1]沈阳军区联勤部疾病预防控制中心,辽宁省沈阳市110034 [2]沈阳药科大学 [3]吉林大学第一医院

出　　处：《职业与健康》2011年第21期2417-2419,共3页Occupation and Health

基　　金：长春市科学技术基金项目资助(项目编号:No.04-07SF057)

摘　　要：目的构建艾滋病病毒核心蛋白(gag)与干扰素(IFNα-2b)融合基因表达质粒,观察其与痘苗病毒共表达的结果,研究其意义。方法利用基因重组技术,将IFNα-2b基因片段插入到gag基因的nt 531位点,经脂质体转染与血凝素阴性蚀斑筛选,挑出重组病毒。经免疫荧光、蛋白印迹分析(Western blot)和斑点酶联免疫(Dot ELISA)鉴定表达产物。结果间接免疫荧光实验显示,转染重组质粒的细胞表面有绿色荧光。免疫印迹实验与Dot ELISA结果均显示,转染重组质粒的细胞裂解物中存在表达的gag/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 the expression plasmid of HIV-l gag and IFNα-2b fusion gene,observe the effect of co-expression of it and vaccinia virus,and study its significance. [Methods]By using gene recombination technique,IFNα-2b gene fragment was inserted into nt 531 site of gag gene,and the recombinant virus was selected by liposome transfection and screening of hemagglutinin negative plaque.The expression product was examined by immunofluorescence,Western blot and Dot ELISA. [Results]The indirect immunofluorescence assay showed that there was green fluorescence in the surface of cell with recombinant plasmid.The results of Western blot and Dot ELISA showed that there was gag/IFNα-2b protein in lysates of cell with recombinant plasmid. [Conclusion]The recombinant eukaryotic cells expression plasmid is successfully constructed,and expression protein has good immunogenicity and immune reactivity.

关 键 词：痘苗病毒 免疫原 重组质粒 艾滋病 

分 类 号：R392[医药卫生—免疫学]