title: Transient dominant host-range selection using Chinese hamster ovary cells to generate marker-free recombinant viral vectors from vaccinia virus.
abstract: Recombinant vaccinia viruses (rVACVs) are promising antigen-delivery systems for vaccine development that are also useful as research tools. Two common methods for selection during construction of rVACV clones are (i) co-insertion of drug resistance or reporter protein genes, which requires the use of additional selection drugs or detection methods, and (ii) dominant host-range selection. The latter uses VACV variants rendered replication-incompetent in host cell lines by the deletion of host-range genes. Replicative ability is restored by co-insertion of the host-range genes, providing for dominant selection of the recombinant viruses. Here, we describe a new method for the construction of rVACVs using the cowpox CP77 protein and unmodified VACV as the starting material. Our selection system will expand the range of tools available for positive selection of rVACV during vector construction, and it is substantially more high-fidelity than approaches based on selection for drug resistance.
authors.affiliation: Experimental Therapeutics Laboratory, Hanson Institute and Sansom Institute, Adelaide, Australia.|School of Pharmacy and Medical Sciences, University of South Australia, Adelaide, Australia.
authors.forename: Liang
authors.initials: L
authors.lastname: Liu
authors.affiliation: Experimental Therapeutics Laboratory, Hanson Institute and Sansom Institute, Adelaide, Australia.|School of Pharmacy and Medical Sciences, University of South Australia, Adelaide, Australia.
authors.forename: Tamara
authors.initials: T
authors.lastname: Cooper
authors.affiliation: Experimental Therapeutics Laboratory, Hanson Institute and Sansom Institute, Adelaide, Australia.|School of Pharmacy and Medical Sciences, University of South Australia, Adelaide, Australia.
authors.forename: Preethi
authors.initials: P
authors.lastname: Eldi
authors.affiliation: Experimental Therapeutics Laboratory, Hanson Institute and Sansom Institute, Adelaide, Australia.|School of Pharmacy and Medical Sciences, University of South Australia, Adelaide, Australia.
authors.forename: Pablo
authors.initials: P
authors.lastname: Garcia-Valtanen
authors.affiliation: Experimental Therapeutics Laboratory, Hanson Institute and Sansom Institute, Adelaide, Australia.|School of Pharmacy and Medical Sciences, University of South Australia, Adelaide, Australia.|Robinson Research Institute and Adelaide Medical School, University of Adelaide, Adelaide, Australia.
authors.forename: Kerrilyn R
authors.initials: KR
authors.lastname: Diener
authors.affiliation: Experimental Therapeutics Laboratory, Hanson Institute and Sansom Institute, Adelaide, Australia.|School of Pharmacy and Medical Sciences, University of South Australia, Adelaide, Australia.|Sementis Ltd., Melbourne, Australia.
authors.forename: Paul M
authors.initials: PM
authors.lastname: Howley
authors.affiliation: Experimental Therapeutics Laboratory, Hanson Institute and Sansom Institute, Adelaide, Australia.|School of Pharmacy and Medical Sciences, University of South Australia, Adelaide, Australia.|Robinson Research Institute and Adelaide Medical School, University of Adelaide, Adelaide, Australia.
authors.forename: John D
authors.initials: JD
authors.lastname: Hayball
mesh_terms: D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D004261:DNA Replication / Q000235:genetics; D004351:Drug Resistance / Q000235:genetics*; D005822:Genetic Vectors; D006801:Humans; D011995:Recombination, Genetic; D017384:Sequence Deletion; D014616:Vaccinia virus / Q000235:genetics*; D014779:Virus Replication / Q000235:genetics
keywords: CHO; CP77; Vaccinia; host-range factor; recombinant virus construction