Characterization of recombinant vaccinia viruses and their products.
After a recombinant vaccinia virus is made, its DNA and protein products can be analyzed in several ways. Protocols are provided in this unit for identification of the recombinant virus by PCR (with verification of correct insertion of the DNA by Southern blotting) and by dot-blot hybridization. Also, when antibodies are available, protein expression can be analyzed by immunological methods detailed here such as dot blotting with an antibody, immunoblotting and/or immunoprecipitation. In addition, immunostaining can be used for identification of recombinant plaques as well as for determination of the purity of a recombinant virus stock. All of the protocols in this unit can be used for characterization of modified vaccinia virus Ankara (MVA) recombinant viruses.
Earl
P L
PL
National Institute of Allergy and Infectious Diseases, Bethesda, Maryland, USA.
Moss
B
B
D004279:DNA, Viral / Q000235:genetics; D047468:Immunoprecipitation; D009693:Nucleic Acid Hybridization / Q000379:methods; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics; D011995:Recombination, Genetic; D014616:Vaccinia virus / Q000235:genetics* / Q000378:metabolism*
D004279:DNA, Viral; D011994:Recombinant Proteins