HIV-1 env基因在重组鸡痘病毒中的表达

在以鸡痘病毒复制非必需区TK基因为侧翼构建的载体 (插入有 16个串联的突变型痘苗病毒P7 5启动子和牛痘病毒A型包涵体晚期启动子及LacZ报告基因 )中的复合启动子下游 ,分别插入编码人免疫缺陷病毒(HIV)Ⅰ型外膜蛋白的env基因和中国株HIV 1gp12 0基因 ,通过与野生型鸡痘病毒的同源重组 ,X gal显色挑斑 ,获得 2株重组病毒vUTA - 2f- 16LE和vUTA - 2f- 16LG。经Dot-ELISA、免疫荧光试验、SDS -PAGE和Westernblot检测表明 ,2株重组病毒均能表达gp16 0和gp12 0 ,相对分子质量分别为 16 0 0 0 0和 12 0 0 0 0。经小鼠免疫试验其血清中抗gp12 0抗体A值和脾脏CD4+ /CD8+ T淋巴细胞比值均高于对照组 (P <0 .0 1) ,说明由重组病毒表达的Env蛋白和Gp12 0蛋白具有良好的反应原性和免疫原性。

Expression of HIV-1 Gene in Recombinant Fowlpox Virus

The env gene encoding the envelope protein type 1 of AIDS virus(HIV) and gp120 gene of Chinese strain of HIV 1 were inserted to the downstream of hybrid promotor of carrier(containing 16 P7.5 promotors of fowlpox virus mutant,late promotor of inclusion body type A of cowpox virus and LacZ labeled gene) constructed using the TK gene at unnecessary regions of fowlpox virus as flank, then 2 strains of recombinant chicken virus vUTA 2f 16LE and vUTA 2f 16LG were obtained by homologous recombination with wild fowlpox virus and selection of blue blot using X gal as substrate.Dot ELISA,IFA,SDS PAGE and Western blot demonstrated that both the viruses could express gp160 and gp120 with the molecular weights of 160 000 and 120 000, respectively. The immunization experiment in mice proved that the A vlue of antibody to gp120 in sera and the ratio of CD4 to CD8 in spleen were significantly higher than those in control group. It showed good reactogenicity and immunogenicity of expressed env and gp120 proteins.