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  • Viral Transfer Technology
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New tools for the generation of E1- and/or E3-substituted adenoviral vectors

Gene Therapy volume 7, pages 80–87 (2000)Cite this article

Abstract

We have designed new vectors for the construction of recombinant adenoviruses containing expression cassettes in the E1 and/or E3 regions. Using a versatile set of restriction enzymes, the cassettes are cloned into small bacterial vectors and subsequently introduced into large plasmids containing the adenoviral sequences. Two positive selection markers facilitate the recovery of a cosmid containing a copy of the sequence of the recombinant adenovirus. The resulting cosmid is transfected into 293 or 911 cells in order to rescue the virus. Importantly, the method does not require any recombination event, either in E. coli or in mammalian cells. The entire procedure can generate viral plaques in 12 days.

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Acknowledgements

We gratefully acknowledge Drs E Chang, Y Laroche and D Salmi for critical review of the manuscript, and Dr RE Vestal for generous financial help.

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Authors and Affiliations

  1. Mountain States Medical Research Institute and Department of Veteran Affairs Medical Center, Boise, ID, USA

    X Danthinne & E Werth

  2. Center for Transgene Technology and Gene Therapy, Vlaams Interuniversitair Instituut voor Biotechnologie, Leuven, Belgium

    X Danthinne

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  1. X Danthinne
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  2. E Werth
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Danthinne, X., Werth, E. New tools for the generation of E1- and/or E3-substituted adenoviral vectors. Gene Ther 7, 80–87 (2000). https://doi.org/10.1038/sj.gt.3301047

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