Engineering recombinant poxviruses using a compact GFP-blasticidin resistance fusion gene for selection

Yik Chun Wong; Leon C.W. Lin; Carolina R. Melo-Silva; Stewart A. Smith; David C. Tscharke

doi:10.1016/j.jviromet.2010.11.003

Engineering recombinant poxviruses using a compact GFP-blasticidin resistance fusion gene for selection

Yik Chun Wong, Leon C.W. Lin, Carolina R. Melo-Silva, Stewart A. Smith, David C. Tscharke^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

29 Citations (Scopus)

Abstract

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.

Original language	English
Pages (from-to)	295-298
Number of pages	4
Journal	Journal of Virological Methods
Volume	171
Issue number	1
DOIs	https://doi.org/10.1016/j.jviromet.2010.11.003
Publication status	Published - Jan 2011

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10.1016/j.jviromet.2010.11.003

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title = "Engineering recombinant poxviruses using a compact GFP-blasticidin resistance fusion gene for selection",

abstract = "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.",

keywords = "Ectromelia virus, Modified Vaccinia Ankara, Poxvirus, Recombinant vaccine, Transient dominant selection, Vaccinia virus",

author = "Wong, {Yik Chun} and Lin, {Leon C.W.} and Melo-Silva, {Carolina R.} and Smith, {Stewart A.} and Tscharke, {David C.}",

year = "2011",

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doi = "10.1016/j.jviromet.2010.11.003",

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AU - Wong, Yik Chun

AU - Lin, Leon C.W.

AU - Melo-Silva, Carolina R.

AU - Smith, Stewart A.

AU - Tscharke, David C.

PY - 2011/1

Y1 - 2011/1

N2 - 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.

AB - 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.

KW - Ectromelia virus

KW - Modified Vaccinia Ankara

KW - Poxvirus

KW - Recombinant vaccine

KW - Transient dominant selection

KW - Vaccinia virus

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JF - Journal of Virological Methods

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Engineering recombinant poxviruses using a compact GFP-blasticidin resistance fusion gene for selection

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