title: Generation of Recombinant Vaccinia Viruses.
abstract: This unit describes how to infect cells with vaccinia virus and then transfect them with a plasmid-transfer vector or PCR fragment to generate a recombinant virus. Selection and screening methods used to isolate recombinant viruses and a method for the amplification of recombinant viruses are described. Finally, a method for live immunostaining that has been used primarily for detection of recombinant modified vaccinia virus Ankara (MVA) is presented.
authors.affiliation: National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland.
authors.forename: Linda S
authors.initials: LS
authors.lastname: Wyatt
authors.affiliation: National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland.
authors.forename: Patricia L
authors.initials: PL
authors.lastname: Earl
authors.affiliation: National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland.
authors.forename: Bernard
authors.initials: B
authors.lastname: Moss
mesh_terms: D005820:Genetic Testing; D007158:Immunologic Techniques / Q000379:methods; D008967:Molecular Biology / Q000379:methods*; D011995:Recombination, Genetic; D012641:Selection, Genetic; D013194:Staining and Labeling / Q000379:methods; D014162:Transfection / Q000379:methods; D014616:Vaccinia virus / Q000235:genetics*; D014773:Virology / Q000379:methods*; D014776:Virus Cultivation / Q000379:methods