论文部分内容阅读
以痘苗病毒复制非必需区血凝素（ＨＡ）基因为侧翼，将编码人白细胞介素２（ｈＩＬ－２）的基因片段克隆到真核表达质粒ｐＪ３８ｅｎｖ 的下游，构建成含有ＨＩＶ－１ ｅｎｖ 和ｈＩＬ－２两种外源基因的重组表达质粒ｐＪ３８Ｅ－ＩＬ－２，经同源重组和血凝素阴性空斑筛选，获得了重组痘苗病毒ｖＪ３８Ｅ－ＩＬ２。经间接免疫荧光试验、Ｄｏｔ－ＥＬＩＳＡ和Ｗｅｓｔｅｒｎ ｂｏｌｔ等检测证明：重组病毒能同时表达Ｅｎｖ 蛋白和ＩＬ－２蛋白，表达产物分子量分别为９８×１０３ 、１６×１０３。
The gene fragment encoding human interleukin 2 (hIL-2) was cloned downstream of the vaccinia virus non-essential hemagglutinin (HA) gene into the downstream of the eukaryotic expression plasmid pJ38env and constructed into a vector containing HIV-1 env and Recombinant vaccinia virus vJ38E-IL2 was obtained by homologous recombination and hemagglutinin-negative plaque screening of the recombinant expression plasmid pJ38E-IL-2 of two foreign genes of hIL-2. The results of indirect immunofluorescence assay, Dot-ELISA and Western bolt showed that the recombinant virus could express Env protein and IL-2 protein at the same time. The molecular weight of the recombinant protein was 98 × 103 and 16 × 103, respectively.