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Journal of Virology, January 2005, p. 486-494, Vol. 79, No. 1
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.1.486-494.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Conditional Cytomegalovirus Replication In Vitro and In Vivo

Max von Pettenkofer Institut für Virologie, Ludwig-Maximilians-Universität München, Munich, Germany

Received 28 May 2004/ Accepted 7 September 2004

We have established a conditional gene expression system for cytomegalovirus which allows regulation of genes independently from the viral replication program. Due to the combination of all elements required for regulated expression in the same viral genome, conditional viruses can be studied in different cell lines in vitro and in the natural host in vivo. The combination of a self-sufficient tetracycline-regulated expression cassette and Flp recombinase-mediated insertion into the viral genome allowed fast construction of recombinant murine cytomegaloviruses carrying different conditional genes. The regulation of two reporter genes, the essential viral M50 gene and a dominant-negative mutant gene (m48.2) encoding the small capsid protein, was analyzed in more detail. In vitro, viral growth was regulated by the conditional expression of M50 by 3 orders of magnitude and up to a millionfold when the dominant-negative small capsid protein mutant was used. In vivo, viral growth of the dominant-negative mutant was reduced to detection limits in response to the presence of doxycycline in the organs of mice. We believe that this conditional expression system is applicable to genetic studies of large DNA viruses in general.

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