Construction and Isolation of Recombinant Vaccinia Virus Using Genetic Markers
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Last Update: 2021-02-22
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Source: Internet
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Author: User
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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.
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