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Journal of Virology, September 1999, p. 7582-7589, Vol. 73, No. 9
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

An Adenovirus-Epstein-Barr Virus Hybrid Vector That Stably Transforms Cultured Cells with High Efficiency

Brenton T. Tan,1 Lily Wu,2 and Arnold J. Berk1,*

Department of Microbiology and Molecular Genetics, Molecular Biology Institute,1 and Department of Urology, School of Medicine,2 University of California, Los Angeles, Los Angeles, California 90095

Received 5 April 1999/Accepted 11 June 1999

EBV episomes are nuclear plasmids that are stably maintained through multiple cell divisions in primate and canine cells (J. L. Yates, N. Warren, and B. Sugden, Nature 313:812-815, 1985). In this report, we describe the construction and characterization of an E1-deleted recombinant adenovirus vector system that delivers an EBV episome to infected cells. This adenovirus-EBV hybrid vector system utilizes Cre-mediated, site-specific recombination to excise an EBV episome from a target recombinant adenovirus genome. We demonstrate that this vector system efficiently delivers the EBV episome and stably transforms a large fraction of infected canine D-17 cells. Using a colony-forming assay, we demonstrate stable transformation of 37% of cells that survive the infection. However, maximal transformation efficiency is achieved at doses of the E1-deleted recombinant adenoviruses that are toxic to the infected cells. Consequently, E1-deleted vector toxicity imposes a limitation on our current vector system.

* Corresponding author. Mailing address: Department of Microbiology and Molecular Genetics, Molecular Biology Institute, University of California, Los Angeles, 611 Charles E. Young Dr. East, Box 951570, Los Angeles, CA 90095-1507. Phone: (310) 206-6298. Fax: (310) 206-7286. E-mail: berk{at}ewald.mbi.ucla.edu.

Journal of Virology, September 1999, p. 7582-7589, Vol. 73, No. 9
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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