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      Efficient generation of recombinant adenovirus vectors by homologous recombination in Escherichia coli.

      Journal of Biology
      Adenoviruses, Human, genetics, Cloning, Molecular, Escherichia coli, Genetic Vectors, Humans, Recombination, Genetic

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          Abstract

          Despite recent technical improvements, the construction of recombinant adenovirus vectors remains a time-consuming procedure which requires extensive manipulations of the viral genome in both Escherichia coli and eukaryotic cells. This report describes a novel system based on the cloning and manipulation of the full-length adenovirus genome as a stable plasmid in E. coli, by using the bacterial homologous recombination machinery. The efficiency and flexibility of the method are illustrated by the cloning of the wild-type adenovirus type 5 genome, the insertion of a constitutive promoter upstream from the E3 region, the replacement of the E1 region by an exogenous expression cassette, and the deletion of the E1 region. All recombinant viral DNAS were shown to be fully infectious in permissive cells, and the modified E3 region or the inserted foreign gene was correctly expressed in the infected cells.

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          Author and article information

          Journal
          8676512
          190422
          10.1128/jvi.70.7.4805-4810.1996

          Chemistry
          Adenoviruses, Human,genetics,Cloning, Molecular,Escherichia coli,Genetic Vectors,Humans,Recombination, Genetic

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