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Journal of Virology, March 2005, p. 2998-3008, Vol. 79, No. 5
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.5.2998-3008.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Use of a Murine Cytomegalovirus K181-Derived Bacterial Artificial Chromosome as a Vaccine Vector for Immunocontraception

Alec J. Redwood,1,2* Martin Messerle,3,4 Nicole L. Harvey,1,2 Christopher M. Hardy,2 Ulrich H. Koszinowski,3 Malcolm A. Lawson,1 and Geoffrey R. Shellam1

Microbiology and Immunology, School of Biomedical and Chemical Sciences, The University of Western Australia, Crawley, Western Australia,1 Pest Animal Control Cooperative Research Centre, CSIRO Sustainable Ecosystems, Canberra, Australian Capital Territory, Australia,2 Department of Virology, Max von Pettenkofer-Institute, Genzentrum, Ludwig-Maximilians-University of Munich, Munich,3 Virus-Cell-Interaction Unit, Medical Faculty, University of Halle-Wittenberg, Halle (Saale), Germany4

Received 15 March 2004/ Accepted 21 September 2004

Cytomegaloviruses (CMVs) are members of the Betaherpesvirinae subfamily of the Herpesviridae, and their properties of latency, large DNA size, gene redundancy, and ability to be cloned as bacterial artificial chromosomes (BACs) suggest their utility as vaccine vectors. While the K181 strain of murine CMV (MCMV) is widely used to study MCMV biology, a BAC clone of this virus had not previously been produced. We report here the construction of a BAC clone of the K181Perth strain of MCMV. The in vivo and in vitro growth characteristics of virus derived from the K181 BAC were similar to those of wild-type K181. The utility of the K181 BAC as a method for the rapid production of vaccine vectors was assessed. A vaccine strain of BAC virus, expressing the self-fertility antigen, murine zona pellucida 3, was produced rapidly using standard bacterial genetics techniques and rendered female BALB/c mice infertile with a single intraperitoneal inoculation. In addition, attenuated vaccine strains lacking the open reading frames m07 to m12 exhibited no reduction in efficacy compared to the full-length vaccine strain. In conclusion, we describe the production of a K181-based BAC virus which behaved essentially as wild-type K181 and allowed the rapid production of effective viral vaccine vectors.

* Corresponding author. Mailing address: Microbiology and Immunology, School of Biomedical and Chemical Sciences, M502, The University of Western Australia, 35 Stirling Highway, Crawley, Western Australia 6009, Australia. Phone: 61 08 9346 2512. Fax: 61 08 9346 2912. E-mail: aredwood{at}cyllene.uwa.edu.au.

Journal of Virology, March 2005, p. 2998-3008, Vol. 79, No. 5
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.5.2998-3008.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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