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A novel adenovirus vector for easy cloning in the E3 region downstream of the CMV promoter
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...
Autores principales: | , , , |
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Formato: | Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2008
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2427025/ https://www.ncbi.nlm.nih.gov/pubmed/18538014 http://dx.doi.org/10.1186/1743-422X-5-73 |
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author | Mailly, Laurent Boulade-Ladame, Charlotte Orfanoudakis, Georges Deryckere, François |
author_facet | Mailly, Laurent Boulade-Ladame, Charlotte Orfanoudakis, Georges Deryckere, François |
author_sort | Mailly, Laurent |
collection | PubMed |
description | 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. |
format | Text |
id | pubmed-2427025 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2008 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-24270252008-06-13 A novel adenovirus vector for easy cloning in the E3 region downstream of the CMV promoter Mailly, Laurent Boulade-Ladame, Charlotte Orfanoudakis, Georges Deryckere, François Virol J Short Report 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. BioMed Central 2008-06-06 /pmc/articles/PMC2427025/ /pubmed/18538014 http://dx.doi.org/10.1186/1743-422X-5-73 Text en Copyright © 2008 Mailly et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 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. |
spellingShingle | Short Report Mailly, Laurent Boulade-Ladame, Charlotte Orfanoudakis, Georges Deryckere, François A novel adenovirus vector for easy cloning in the E3 region downstream of the CMV promoter |
title | A novel adenovirus vector for easy cloning in the E3 region downstream of the CMV promoter |
title_full | A novel adenovirus vector for easy cloning in the E3 region downstream of the CMV promoter |
title_fullStr | A novel adenovirus vector for easy cloning in the E3 region downstream of the CMV promoter |
title_full_unstemmed | A novel adenovirus vector for easy cloning in the E3 region downstream of the CMV promoter |
title_short | A novel adenovirus vector for easy cloning in the E3 region downstream of the CMV promoter |
title_sort | novel adenovirus vector for easy cloning in the e3 region downstream of the cmv promoter |
topic | Short Report |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2427025/ https://www.ncbi.nlm.nih.gov/pubmed/18538014 http://dx.doi.org/10.1186/1743-422X-5-73 |
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