Abstract
The construction of deletion-knockout poxviruses is a useful approach to determining the function of specific virus genes. This protocol is an adaptation of the transient dominant knockout selection protocol published by Falkner and Moss (1990) for use with vaccinia virus. The protocol makes use of the dominant selectable marker Escherichia coli guanine phosphoribosyltransferase (gpt) gene (Mulligan and Berg, 1981), under the control of an early/late poxvirus promoter. The deletion viruses that are produced no longer contain a selectable marker, which may be preferable for the production of vaccines.
Keywords: Knockout, Fowlpox, Deletion, Poxvirus, Transient dominant
Materials and Reagents
Equipment
Procedure
Recipes
Acknowledgments
This protocol was adapted from Boulanger et al. (1998); Falkner and Moss (1990); and Laidlaw et al. (2013). This work was funded by the Biotechnology and Biological Sciences Research Council (BBSRC), grants BBS/B/00115/2, BB/E009956/1, and BB/G018545/1.
References
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