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Journal of Virology, August 1999, p. 7056-7060, Vol. 73, No. 8
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Systematic Excision of Vector Sequences from the BAC-Cloned Herpesvirus Genome during Virus Reconstitution

Markus Wagner,1 Stipan Jonjic,2 Ulrich H. Koszinowski,1,* and Martin Messerle1

Max von Pettenkofer-Institut für Hygiene und Medizinische Mikrobiologie, Ludwig-Maximilians-Universität München, D-81377 Munich, Germany,1 and Department of Embryology and Histology, Faculty of Medicine, University of Rijeka, 51000 Rijeka, Croatia2

Received 8 February 1999/Accepted 14 May 1999

Recently the mouse cytomegalovirus (MCMV) genome was cloned as an infectious bacterial artificial chromosome (BAC) (M. Messerle, I. Crnkovic', W. Hammerschmidt, H. Ziegler, and U. H. Koszinowski, Proc. Natl. Acad. Sci. USA 94:14759-14763, 1997). The virus obtained from this construct is attenuated in vivo due to deletion of viral sequences and insertion of the BAC vector. We reconstituted the full-length MCMV genome and flanked the BAC vector with identical viral sequences. This new construct represents a versatile basis for construction of MCMV mutants since virus generated from the construct loses the bacterial sequences and acquires wild-type properties.

* Corresponding author. Mailing address: Max von Pettenkofer-Institut, Pettenkoferstr. 9a, D-80336 Munich, Germany. Phone: 49 89 5160 5290. Fax: 49 89 5160 5292. E-mail: koszinowski{at}m3401.mpk.med.uni-muenchen.de.

Journal of Virology, August 1999, p. 7056-7060, Vol. 73, No. 8
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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