非复制重组痘苗病毒天坛株C-K缺失区表达载体的构建及重组病毒生物学性状的研究  

Construction of Expression Vectors of Non-replicating Recombinant Vaccinia Virus Tiantan Strain With CK Region Deletion and Studying of Biological Properties of the Recombinant Viruses

作　　者：郭斐[1] 陆柔剑[1] 娄元梅[1] 孙朝晖[1] 阮力[1] 

机构地区：[1]中国预防医学科学院病毒学研究所,北京100052

出　　处：《病毒学报》2001年第1期24-28,共5页Chinese Journal of Virology

基　　金：国家863高技术研究发展计划! (863 -10 2 -0 7-0 2 -0 1) ;国家自然科学基金! (3 95 2 5 0 0 1) ;国家自然科学基金! (3 0 0 70 71

摘　　要：将反向串联的痘苗病毒启动子 p11k和 p7.5k表达结构引入非复制痘苗病毒质粒载体 ,得到非复制痘苗病毒表达载体 pNEOCK11β75、pNEOCK75 11β、pNEOCK1175和 pNEOCK75 11。利用载体 pNEOCK11β75、pNEOCK75 11β及 pNEOCK11β75IL6和RVJ12 3重组病毒进行同源重组 ,分别得到非复制重组痘苗病毒RVJ12 3Δ11β75、RVJ12 3Δ75 11β及RVJ12 3Δ11β75IL6。Southern blot证实 :重组病毒RVJ12 3Δ11β75C K片段间大片段基因的稳定缺失 ,同时 β 半乳糖苷酶基因插入到缺失区内并稳定表达 ,不影响另外非必需区外源基因的表达 ,缺失区内两外源基因的表达无相互干扰 。We inserted expression cassette downstream of vaccinia virus promoters p11k and p7.5k into the constructed non-replicating vaccinia virus vector plasmid, giving rise a set of non-replicating vaccinia virus expression vector plasmids pNEOCK11β75, pNEOCK7511β, pNEOCK1175, and pNEOCK7511. Then we constructed the non-replicating recombinant vaccinia viruses RVJ123ΔCK11β75, RVJ123ΔCK7511β and RVJ123Δ11β75IL6 through RVJ123 recombination with expression vector pNEOCK11β75, pNEOCK7511β and pNEOCK11β75IL6. By Southern-blot, it is determined that the genes between C and K fragments of the non-replicating recombinant viruses are deleted stably and β-gal gene is integrated stably in the deletion region. The inserted gene can express stably and its expression has no effect on the exogenous gene in other non-essential region. The transcription direction of the exogenous gene has no effect on its DNA replication, protein translation and gene expression in other non-essential region.

关 键 词：痘苗病毒 天坛株 非复制表达载体 遗传稳定性 构建 

分 类 号：R373.12[医药卫生—病原生物学] Q78[医药卫生—基础医学]