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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.

Original publication

DOI

10.1002/0471142727.mb1617s43

Type

Journal article

Journal

Current protocols in molecular biology

Publication Date

05/2001

Volume

Chapter 16

Addresses

National Institute of Allergy and Infectious Diseases, Bethesda, Maryland, USA.

Keywords

Cells, Cultured, Cell Line, Chick Embryo, Animals, Humans, Vaccinia virus, Concanavalin A, DNA, Viral, Transfection, Recombination, Genetic, Genetic Vectors