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Vol. 95, Issue 5, 2509-2514, March 3, 1998

Medical Sciences
A simplified system for generating recombinant adenoviruses

Tong-Chuan He^*, Shibin Zhou^*, Luis T. da Costa, Jian Yu, Kenneth W. Kinzler, and Bert Vogelstein^*,,§

^* The Howard Hughes Medical Institute,  The Program in Human Genetics and Molecular Biology,  The Johns Hopkins Oncology Center, 424 North Bond Street, Baltimore, MD 21231

Contributed by Bert Vogelstein, December 30, 1997

Recombinant adenoviruses provide a versatile system for gene expression studies and therapeutic applications. We report herein a strategy that simplifies the generation and production of such viruses. A recombinant adenoviral plasmid is generated with a minimum of enzymatic manipulations, using homologous recombination in bacteria rather than in eukaryotic cells. After transfections of such plasmids into a mammalian packaging cell line, viral production is conveniently followed with the aid of green fluorescent protein, encoded by a gene incorporated into the viral backbone. Homogeneous viruses can be obtained from this procedure without plaque purification. This system should expedite the process of generating and testing recombinant adenoviruses for a variety of purposes.

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