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 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.
Mailly
Laurent
L
Unité Mixte de Recherche 7175, Ecole Supérieure de Biotechnologie de Strasbourg, Illkirch, France. Laurent.Mailly@viro-ulp.u-strasbg.fr
Boulade-Ladame
Charlotte
C
Orfanoudakis
Georges
G
Deryckere
François
F
D017503:Adenovirus E3 Proteins / Q000235:genetics*; D000260:Adenoviruses, Human / Q000235:genetics; D003001:Cloning, Molecular; D015870:Gene Expression; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors / Q000191:economics / Q000235:genetics*; D017874:Immediate-Early Proteins / Q000235:genetics*; D011401:Promoter Regions, Genetic; D019076:Transgenes; D014764:Viral Proteins / Q000235:genetics*
D017503:Adenovirus E3 Proteins; C088645:IE1 protein, cytomegalovirus; D017874:Immediate-Early Proteins; D014764:Viral Proteins