A simplified system for generating recombinant adenoviruses |
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Authors: | TC He S Zhou LT da Costa J Yu KW Kinzler B Vogelstein |
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Affiliation: | The Howard Hughes Medical Institute, 424 North Bond Street, Baltimore, MD 21231, USA. tche@welchlink.welch.jhu.edu |
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Abstract: | 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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