SpringerProtocols: Abstract: Construction and Isolation of Recombinant Vaccinia Virus Using Genetic Markers

Construction and Isolation of Recombinant Vaccinia Virus Using Genetic Markers

By: Maríia M. Lorenzo², Inmaculada Galindo², Rafael Blasco²

Abstract

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The standard approach for the isolation of vaccinia virus recombinants involves homologous recombination between a transfected plasmid and the replicating viral DNA. In a typical infection/transfection experiment, recombinant viruses only account for a tiny proportion (10⁻⁴ to 10⁻³) of the progeny virus; thus, genetic markers are often included in the transfected plasmid to facilitate the selection of recombinant viruses. This chapter describes in detail two different selection procedures: one relies on plaque formation phenotype using the vaccinia virus gene F13L; the other relies on antibiotic resistance using the Escherichia coli xanthine-guanine phosphoribosyl transferase gene.

Affiliation(s): (2) Departamento de Biotecnologíia, Instituto Nacional de Investigacióon y Tecnologíia Agraria y Alimentaria (INIA), Madrid, Spain

Book Title: Vaccinia Virus and Poxvirology: Methods and Protocols

Series: Methods in Molecular Biology | Volume: 269 | Pub. Date: Apr-19-2004 | Page Range: 15-30 | DOI: 10.1385/1-59259-789-0:015

Subject: Microbiology

Key Words: Recombinant vaccinia virus - plaque size - F13L - xanthine-guanine phosphoribosyl - transferase - gpt

References

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