Wyatt L S, Moss B, Rozenblatt S
Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892-0455, USA.
Virology. 1995 Jun 20;210(1):202-5. doi: 10.1006/viro.1995.1332.
The vaccinia virus/bacteriophage T7 hybrid transient expression system employs a recombinant vaccinia virus that encodes the T7 RNA polymerase gene, a plasmid vector with a gene of interest regulated by a T7 promoter, and any cell line suitable for infection and transfection. Although high expression in a majority of cells is achieved, the severe cytopathic effects of vaccinia virus and the safety precautions required for use of infectious agents are undesirable features of the system. Here, we report the construction of a highly attenuated and avian host-restricted vaccinia virus recombinant that encodes the T7 RNA polymerase gene (MVA/T7 pol) and demonstrate the use of the virus for transient expression in mammalian cells. MVA/T7 pol has reduced cytopathic effects compared to the previously used replication-competent vaccinia virus, while providing a high level of gene expression in multiple mammalian cell lines.
痘苗病毒/噬菌体T7杂交瞬时表达系统采用一种编码T7 RNA聚合酶基因的重组痘苗病毒、一个带有受T7启动子调控的目的基因的质粒载体，以及任何适合感染和转染的细胞系。尽管该系统能在大多数细胞中实现高表达，但痘苗病毒严重的细胞病变效应以及使用感染性因子所需的安全预防措施是该系统的不利特征。在此，我们报告构建了一种编码T7 RNA聚合酶基因的高度减毒且宿主受限的痘苗病毒重组体（MVA/T7 pol），并展示了该病毒在哺乳动物细胞中的瞬时表达应用。与之前使用的具有复制能力的痘苗病毒相比，MVA/T7 pol的细胞病变效应有所降低，同时能在多种哺乳动物细胞系中实现高水平的基因表达。
