New vectors for the construction of double recombinant adenoviruses

A set of plasmids designed for the construction of recombinant adenoviral v ectors is described, which contain two expression cassettes, one in the ear ly region 1 (E1) and the other in the early region 3 (E3). Two cloning step s in E. coli and a transfection of the resulting cosmid into 293 cells are sufficient to recover the recombinant virus. The method has been optimised to facilitate the introduction of the genes of interest in their respective regions and the reconstitution of the entire sequence of the recombinant a denoviral DNA in E. coli. The vectors are easy to handle and generate homog enous virus preparations. To illustrate the efficiency of the method, an ad enovirus was constructed expressing the E, coli beta-galactosidase Delta M1 5 mutant in the E1 region and the complementing lacZ ct-peptide in the E3 r egion. (C) 1999 Elsevier Science B.V. All rights reserved.