Production of recombinant adenovirus, useful e.g. in gene therapy and genomics, by homologous recombination between parent and shuttle plasmids

摘要

Production of adenovirus (AdV) by homologous recombination between shuttle plasmid (P1) and parent plasmid (P2) to produce a final plasmid (P3) containing a complete, recombinant AdV genome. This genome is excised as a linear sequence and inserted into a prokaryotic packaging cells to produce AdV containing a complete genome. Production of adenovirus (AdV) by homologous recombination between shuttle plasmid (P1) and parent plasmid (P2) to produce a final plasmid (P3) containing a complete, recombinant AdV genome. This genome is excised as a linear sequence and inserted into a prokaryotic packaging cells to produce AdV containing a complete genome. P1, preferably prokaryotic, contains a truncated AdV genome and at least one heterologous nucleic acid (I) and P2 contains a truncated AdV genome, complementary to that in P1. Independent claims are also included for the following: (a) P1 that contains an AdV ITR (inverted terminal repeat), (I) (or a site for its introduction) and an AdV homology sequence, with ITR flanked by a restriction site not present in the AdV genome; (b) nucleic acid library cloned in P1; (c) P2; (d) cells containing one or more of P1 and P2; (e) a method for preparing AdV libraries; (f) AdV libraries produced by method (e) comprising AdV containing nucleic acid inserts from: (i) library of cDNA, genomic DNA or synthetic DNA; (ii) a library of (semi-)random oligonucleotides; or (iii) a library of sequences able to bind to DNA; and (g) kits containing P1 and P2.