Research School of Biology, The Australian National University, Canberra, ACT, Australia.
J Virol Methods. 2011 Jan;171(1):295-8. doi: 10.1016/j.jviromet.2010.11.003. Epub 2010 Nov 10.
Recombinant poxviruses are important tools for research and some are candidate vaccines. To make these viruses a simple, small vector that can be used to engineer multiple strains of vaccinia virus and other model poxviruses, including ectromelia virus is of value. Here a set of plasmids and methods for making these viruses that uses an enhanced green fluorescent protein-blasticidin resistance (GFP-bsd) fusion gene as a transient selectable marker are described. This gene is smaller than any of the bi-functional selection markers used previously. The versatility of the method across different poxviruses is demonstrated by engineering changes into multiple loci of the WR and Modified Vaccinia Ankara (MVA) strains of vaccinia virus and also ectromelia virus. Finally, a set of vaccinia virus sequences for directing homologous recombination that are very highly conserved was designed and tested. These sequences allow a single plasmid to be used to insert a transgene into multiple strains of the virus.
重组痘病毒是研究的重要工具，有些是候选疫苗。为了使这些病毒成为一种简单的小载体，能够用于工程改造多种痘苗病毒和其他模式痘病毒，包括疱疹病毒，这是有价值的。这里描述了一组质粒和方法，用于制造这些病毒，这些病毒使用增强型绿色荧光蛋白-博来霉素抗性（GFP-bsd）融合基因作为瞬时选择标记。这个基因比以前使用的任何双功能选择标记都要小。该方法在不同痘病毒中的多功能性通过对痘苗病毒的 WR 和改良安卡拉痘苗（MVA）株以及疱疹病毒的多个基因座进行工程改造得到证明。最后，设计并测试了一组用于同源重组的高度保守的痘苗病毒序列。这些序列允许使用单个质粒将转基因插入病毒的多个株系中。
