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.

journal_name

Biotechniques

journal_title

BioTechniques

authors

Liu L,Cooper T,Eldi P,Garcia-Valtanen P,Diener KR,Howley PM,Hayball JD

doi

10.2144/000114537

subject

Has Abstract

pub_date

2017-04-01 00:00:00

pages

183-187

issue

4

eissn

0736-6205

issn

1940-9818

pii

000114537

journal_volume

62

pub_type

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