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Copyright © 2008 Mailly et al; licensee BioMed Central Ltd. A novel adenovirus vector for easy cloning in the E3 region downstream of the CMV promoter Laurent Mailly,1,2 Charlotte Boulade-Ladame,1 Georges Orfanoudakis,1 and François Deryckere  11Unité Mixte de Recherche 7175, Ecole Supérieure de Biotechnologie de Strasbourg, Illkirch, France 2Unité Inserm 748, Institut de Virologie, Strasbourg, France Corresponding author.Laurent Mailly: Laurent.Mailly/at/viro-ulp.u-strasbg.fr; Charlotte Boulade-Ladame: boulade/at/esbs.u-strasbg.fr; Georges Orfanoudakis: orfanoudakis/at/esbs.u-strasbg.fr; François Deryckere: derycker/at/esbs.u-strasbg.fr Received April 4, 2008; Accepted June 6, 2008. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. | ||||
Abstract The construction of expression vectors derived from the human adenovirus type 5 (Ad5), usually based on homologous recombination, is time consuming as a shuttle plasmid has to be selected before recombination with the viral genome. Here, we describe a method allowing direct cloning of a transgene in the E3 region of the Ad5 genome already containing the immediate early CMV promoter upstream of three unique restriction sites. This allowed the construction of recombinant adenoviral genomes in just one step, reducing considerably the time of selection and, of course, production of the corresponding vectors. Using this vector, we produced recombinant adenoviruses, each giving high-level expression of the transgene in the transduced cells. | ||||
