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利用重组质粒 pNeo-CK与 pNeo-CKLacZ和表达乙型肝炎 (乙肝 )病毒S抗原的重组痘苗病毒RVJ12 3[1] ,构建了非复制型重组痘苗病毒RVJ12 3ΔCK。Southernblot证实 ,非复制型重组病毒RVJ12 3ΔCK基因组C和K片段间与宿主范围和毒力相关的基因稳定缺失 ,同时 ,J片段中插入的乙肝S抗原基因稳定存在。重组病毒RVJ12 3ΔCK在鸡胚成纤维母细胞中可良好繁殖 ,而在人源细胞系中不繁殖或仅低度繁殖 ,但都能表达HBsAg ,并且在病毒一个复制周期内 ,复制型和非复制型病毒HBsAg表达水平无明显差别。
The non-replicating recombinant vaccinia virus RVJ12 3ΔCK was constructed by using the recombinant plasmids pNeo-CK and pNeo-CKLacZ and the recombinant vaccinia virus RVJ12 3 expressing the hepatitis B virus (S) antigen [1]. Southernblot showed that the host-specific and virulence-related genes of non-replicating RVJ12 3ΔCK genomic C and K fragments were stably deleted and the hepatitis B S antigen gene inserted into J fragment was stable. The recombinant virus RVJ12 3ΔCK reproduces well in chicken embryo fibroblasts but does not or only rarely propagate in human cell lines but both express HBsAg and replicate and non-replicate within one replication cycle of the virus The expression level of HBsAg did not differ significantly.