Engineering recombinant poxviruses using a compact GFP-blasticidin resistance fusion gene for selection
Engineering recombinant poxviruses using a compact GFP-blasticidin resistance fusion gene for selection
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DOI：
10.1016/j.jviromet.2010.11.003
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发表时间：
2011-01-01
影响因子：
3.1
通讯作者：
Tscharke, David C.
中科院分区：
文献类型：
--
作者：
Wong, Yik Chun;Lin, Leon C. W.;Tscharke, David C.
重组痘病毒是研究的重要工具，有些是候选疫苗。使这些病毒成为一种简单、小的载体，可用于工程化多株牛痘病毒和其他模式痘病毒，包括鼠痘病毒，是有价值的。本文描述了一组质粒和制备这些病毒的方法，所述病毒使用增强的绿色荧光蛋白-杀稻瘟菌素抗性（GFP-bsd）融合基因作为瞬时选择标记。该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通用性通过将改变工程化到牛痘病毒的WR和改良的安卡拉牛痘（MVA）株以及脱肢病病毒的多个基因座中来证明。最后，设计并测试了一组非常高度保守的用于指导同源重组的牛痘病毒序列。这些序列允许使用单个质粒将转基因插入病毒的多个毒株中。(C)2010 Elsevier B. V.保留所有权利。
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. (C) 2010 Elsevier B.V. All rights reserved.