Peeters Ben, de Leeuw Olav
Wageningen Bioveterinary Research, Department of Virology, Houtribweg 39, 8221 RA Lelystad, The Netherlands.
J Virol Methods. 2017 Oct;248:187-190. doi: 10.1016/j.jviromet.2017.07.008. Epub 2017 Jul 22.
Reverse genetics systems for non-segmented negative-strand RNA viruses rely on co-transfection of a plasmid containing the full-length viral cDNA and helper plasmids encoding essential viral replication proteins. Here, a system is presented in which virus can be rescued from a single plasmid without the need for helper plasmids in cells infected with a host-restricted recombinant poxvirus that expresses T7 RNA polymerase. This approach relies on the insertion of T7 promoter sequences in the viral cDNA at positions that allow transcription of sub-genomic RNAs encoding essential viral replication proteins.
非节段性负链RNA病毒的反向遗传学系统依赖于共转染一个含有全长病毒cDNA的质粒和编码必需病毒复制蛋白的辅助质粒。在此，我们提出了一种系统，在感染了表达T7 RNA聚合酶的宿主限制性重组痘病毒的细胞中，无需辅助质粒，病毒就可以从单个质粒中拯救出来。这种方法依赖于在病毒cDNA中允许转录编码必需病毒复制蛋白的亚基因组RNA的位置插入T7启动子序列。
