Construction and isolation of recombinant vaccinia virus using genetic markers

Methods Mol Biol. 2004:269:15-30. doi: 10.1385/1-59259-789-0:015.

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.

Publication types

  • Review

MeSH terms

  • Animals
  • DNA, Recombinant*
  • Genetic Markers / genetics*
  • Genetic Vectors
  • Hypoxanthine Phosphoribosyltransferase / genetics
  • Vaccinia virus / genetics*
  • Vaccinia virus / immunology
  • Vaccinia virus / isolation & purification

Substances

  • DNA, Recombinant
  • Genetic Markers
  • Hypoxanthine Phosphoribosyltransferase