期刊
JOURNAL OF VIROLOGICAL METHODS
卷 171, 期 1, 页码 295-298出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jviromet.2010.11.003
关键词
Poxvirus; Vaccinia virus; Modified Vaccinia Ankara; Ectromelia virus; Recombinant vaccine; Transient dominant selection
资金
- Australian NHMRC (Biomedical CDA) [418108, 389819, 509199]
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.
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