ABSTRACT We describe here a clonal approach for efficient and robust construction of recombinant adenoviral genomes that holds certain advantages over existing approaches. Transgenes of

interest are cloned into a small, conditionally replicating plasmid containing the left end of a recombinant adenoviral genome, encompassing pIX coding regions. Transformation of this

plasmid into recombination-competent Escherichia coli bearing a plasmid containing the right end of a recombinant adenoviral genome, commencing from pIX coding regions, yields a stable

co-integrated plasmid encoding a full adenoviral genome, by virtue of shared homology in pIX coding regions contained in both plasmids. The recombination process yielding the full adenoviral

We have successfully employed this approach to generate over 200 recombinant adenoviruses, obtaining only the desired recombinant adenoviral species each time. The process is amenable to

medium-to-high-throughput parallel construction of adenoviral genomes, and as such should aid efforts aimed towards high-throughput functional annotation of therapeutic gene targets, which

aim to leverage the benefits of adenoviruses as gene delivery and expression vectors. Access through your institution Buy or subscribe This is a preview of subscription content, access via

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FREQUENT CONCATEMERIC INSERTIONS OF VIRAL VECTORS Article 08 April 2024 HIGH-THROUGHPUT 5′ UTR ENGINEERING FOR ENHANCED PROTEIN PRODUCTION IN NON-VIRAL GENE THERAPIES Article Open access 06

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acknowledge the support of the Centre National de la Recherche Scientifique and the European Union Marie Curie Fellowship Programme in carrying out this study. We are also grateful to

Patrice Yeh for reviewing the manuscript prior to submission. AUTHOR INFORMATION AUTHORS AND AFFILIATIONS * Aventis Pharma, Functional Genomics, Vitry-sur-Seine Cedex, France B Mullan, C

Dugué, V Moutard, D Raoux, G Tremp, P Denèfle & J-J Robert * CNRS UMR 8121, Institut Gustave Roussy, Villejuif Cedex, France B Mullan, C Dugué, V Moutard, D Raoux, M Perricaudet & 

J-J Robert Authors * B Mullan View author publications You can also search for this author inPubMed Google Scholar * C Dugué View author publications You can also search for this author

inPubMed Google Scholar * V Moutard View author publications You can also search for this author inPubMed Google Scholar * D Raoux View author publications You can also search for this

author inPubMed Google Scholar * G Tremp View author publications You can also search for this author inPubMed Google Scholar * P Denèfle View author publications You can also search for

this author inPubMed Google Scholar * M Perricaudet View author publications You can also search for this author inPubMed Google Scholar * J-J Robert View author publications You can also

search for this author inPubMed Google Scholar RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Mullan, B., Dugué, C., Moutard, V. _et al._ Robust

functional gene validation by adenoviral vectors: one-step Escherichia coli-Derived Recombinant Adenoviral Genome construction. _Gene Ther_ 11, 1599–1605 (2004).

https://doi.org/10.1038/sj.gt.3302333 Download citation * Received: 21 March 2004 * Accepted: 11 June 2004 * Published: 26 August 2004 * Issue Date: 01 November 2004 * DOI:

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currently available for this article. Copy to clipboard Provided by the Springer Nature SharedIt content-sharing initiative KEYWORDS * recombinant adenovirus * homologous recombination * E.

coli * gene validation