Construction and isolation of recombinant vaccinia virus using genetic markers.
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.
Lorenzo
María M
MM
Departamento de Biotecnología, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Madrid, Spain.
Galindo
Inmaculada
I
Blasco
Rafael
R
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
D004274:DNA, Recombinant; D005819:Genetic Markers; D007041:Hypoxanthine Phosphoribosyltransferase