在重组痘苗病毒中共表达HIV-1env与hIL-6基因

COEXPRESSION OF HIV-1 ENV AND IL-6 GENE IN RECOMBINANT VACCINIA VIRUS

本实验以 HIV- 1env与 h IL - 6基因在重组痘苗病毒中的共表达研究为目的 ,将痘苗病毒复制非必需区血凝素(HA )基因作为侧翼 ,把编码人白细胞介素 6 (h IL - 6 )的基因片段克隆到真核表达质粒 p J38env的下游 ,构建成含有 HIV- 1env与 h IL - 6两种外源基因的重组表达质粒 PJ38E- IL 6 ,经同源重组和血凝素阴性空斑筛选 ,获得了重组痘苗病毒 v J38E- IL 6。经间接免疫荧光试验、Dot- EL ISA和 Western bolt等检测证明 ,重组病毒能同时表达 env蛋白和 IL - 6蛋白 ,表达产物的分子量分别为 98k Da、2 6 k

In order to coexpress the HIV 1 env and IL 6 gene in recombinant vaccinia with flank sequence of HA gene of vaccinia virus, coexpression plasmid pJ38E IL 6 containing cDNA of human immunodeficiency virus type I envelope gene was constructed by inserting the hIL 6 gene under the pJ38 env plasmid vector. After homologous recombination in Cos 7 cell, recombinant vaccinia viruses were purified in BHK21 cell by using plaque assay. Indirect immunofluorescence assay(IFA),Dot ELISA and Western blot showed that the recombinant vaccinia virus had coexpressed the HIV 1 env and IL 6 protein in transfected BHK21 and the molecular weight were about 98kDa and 26kDa respectively.