Patent ID: 6110735
Filing Date: 2000-08-29
Classification: A61K,A61P,C12N

Abstract:
A method for preparing, in a prokaryotic cell, a recombinant adenoviral vector into the genome of which an exogenous DNA sequence is inserted, by intermolecular homologous recombination comprising the steps of(a) introducing into said prokaryotic cell:(i) a first DNA fragment comprising all of said genome of the adenovirus or said adenoviral genome lacking all or part of the E1 region, E2 region, E3 region, E4 region or a combination thereof, and(ii) a second DNA fragment comprising said exogenous DNA sequence surrounded by flanking sequences A and B which are homologous to sequences of (i) not including sequences of the 5'ITR, 3'ITR and encapsidation regions of said adenoviral genome, such as to allow intermolecular homologous recombination; wherein the first and second DNA fragments are not carried by a single vector, and(b) culturing the prokaryotic cell obtained in (a) under suitable culture conditions to allow homologous recombination and insertion of the exogenous DNA into the first DNA fragment to occur, and recovering the resulting recombinant adenoviral vector.