Abstract
This chapter presents methods for the generation and analysis of recombinant vaccinia viruses. Vaccinia virus is an extremely versatile virus expression system that can be easily manipulated in the laboratory. The widespread use of vaccinia as a vector system is because of the ease with which recombinant vaccinia viruses containing large insertions of foreign DNA can be generated and because of the authenticity of expression of foreign polypeptides during viral replication. Recombinant vaccinia viruses expressing a wide variety of heterologous genes are used for a number of applications, including (1) analyzing gene regulation, (2) producing biologically active molecules in tissue culture, (3) immunizing animals for antibody production, (4) creating live recombinant vaccines, and (5) elucidating the role of specific antigens in inducing defined immune responses. As additional viral vector improvements are made, the potential of vaccinia as a tool for biological research, as a protein production system and as an immunoprophylactic continues to increase.
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