Generation of recombinant vaccinia viruses.
This unit first describes how to infect cells with vaccinia virus and then transfect them with a plasmid-transfer vector to generate a recombinant virus. Methods are also presented for purifying vaccinia virus and for isolating viral DNA, which can be used during transfection. Also presented are selection and screening methods used to isolate recombinant viruses and a method for the amplification of recombinant viruses. Finally, a method for live immunostaining that has been used primarily for detection of recombinant modified vaccinia virus Ankara (MVA) is presented.
Earl
P L
PL
National Institute of Allergy and Infectious Diseases, Bethesda, Maryland, USA.
Moss
B
B
Wyatt
L S
LS
Carroll
M W
MW
D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D002642:Chick Embryo; D003208:Concanavalin A; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification; D005822:Genetic Vectors; D006801:Humans; D011995:Recombination, Genetic; D014162:Transfection; D014616:Vaccinia virus / Q000235:genetics* / Q000302:isolation & purification
D004279:DNA, Viral; D003208:Concanavalin A