title: Construction and isolation of recombinant vaccinia virus using genetic markers.
abstract: 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.
authors.affiliation: Departamento de Biotecnología, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Madrid, Spain.
authors.forename: María M
authors.initials: MM
authors.lastname: Lorenzo
authors.forename: Inmaculada
authors.initials: I
authors.lastname: Galindo
authors.forename: Rafael
authors.initials: R
authors.lastname: Blasco
mesh_terms: D000818:Animals; D004274:DNA, Recombinant; D005819:Genetic Markers / Q000235:genetics*; D005822:Genetic Vectors; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics; D014616:Vaccinia virus / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification
chemical_list: D004274:DNA, Recombinant; D005819:Genetic Markers; D007041:Hypoxanthine Phosphoribosyltransferase