表达HPV16E6和E7蛋白的非复制型重组痘苗病毒的构建  

Construction of Non-replicating Recombinant Vaccinia Virus Expressing HPV16 E6 and E7 Proteins

作　　者：骆卫锋[1] 韩立群[1] 任皎[1] 田厚文[1] 陆振华 谷淑燕[1] 阮力[1] 

机构地区：[1]中国疾病预防控制中心病毒病预防控制所,北京100052

出　　处：《病毒学报》2002年第3期217-221,共5页Chinese Journal of Virology

基　　金：欧共体合作项目基金 (No ERBICI18-CT97-0 2 3 4) ;国家"九五"科技攻关重点项目基金 ( 96-90 6-0 1-11)

摘　　要：为研制HPV16的治疗性疫苗 ,首先将表达质粒 pJSA1175与非复制型痘苗病毒NTVJTK+ 进行同源重组 ,构建了痘苗重组病毒NTVJLac。再将表达质粒 pJSDME6E7R与NTVJLac进行同源重组 ,构建了表达HPV16E6和E7蛋白的非复制型重组痘苗病毒NTVJmE6E7,并对获得的重组病毒进行了鉴定。Southern杂交显示 ,重组痘苗病毒NTVJmE6E7基因组中有E6和E7基因插入。该重组病毒在人源细胞中不复制。Westernblot显示 ,重组病毒在人源TK-14 3细胞中能表达E6和E7蛋白。To develop antigen-specific theraperutic vaccine for HPV16-associated tumors and their precancerous transformations,a non-replicating recombinant vaccinia virus NTVJLac was constructed through the homologous recombination between vector pJSA1175 and NTVJTK + Then non-replicating recombinant vaccinia virus NTVJmE6E7 was constructed by replacing the LacZ gene from NTVJLac with mE6 and mE7-1 genes of pJSDME6E7R through homologous recombination Southern blot showed mE6 and mE7-1 genes had been inserted in recombinant NTVJmE6E7 NTVJmE6E7 could not replicate in human cells Western blot showed NTVJmE6E7 could express mE6 and mE7 proteins in TK -143 cells NTVJmE6E7 could be taken as a candidate of therapeutic vaccine for HPV-associated tumors and their precancerous transformations

关 键 词：表达 HPV16E6 E7蛋白 非复制型重组痘苗病毒 乳头瘤病毒 宫颈肿瘤 疫苗 免疫治疗 

分 类 号：R373.9[医药卫生—病原生物学] R730.51[医药卫生—基础医学]