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A simplified system for generating recombinant adenoviruses.

T C He·S Zhou·L T da Costa·J Yu·K W Kinzler·B Vogelstein
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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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AdenoviridaeAnimalsCell LineEscherichia coliGenes, ReporterGenetic VectorsGreen Fluorescent ProteinsHumansKidneyLuminescent ProteinsMammalsPlasmidsPolymerase Chain ReactionRecombinant Fusion ProteinsRecombination, GeneticRepetitive Sequences, Nucleic AcidTransfectionJournal ArticleResearch Support, Non-U.S. Gov'tResearch Support, U.S. Gov't, P.H.S.

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T C He, S Zhou, L T da Costa, J Yu, K W Kinzler, B Vogelstein. 1998-03-03. A simplified system for generating recombinant adenoviruses.. https://doi.org/10.1073/pnas.95.5.2509

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