Patent ID: 6303362
Filing Date: 2001-10-16
Classification: A61K,C07K,C12N

Abstract:
A method of preparing a recombinant adenoviral genome having an insertion nucleic acid located in an E gene region of said genome, said method comprising:(a) providing a first vector comprising an adenoviral genome having an E gene region with a deletion of 1.0 to 12.0 kb, where said first vector is further characterized by having first, second and third restriction endonuclease sites, wherein said second restriction endonuclease site is flanked by said first and third restriction endonuclease sites, wherein each of said first, second and third restriction endoncuclease sites are: (i) different, (ii) do not occur in the corresponding wild type adenoviral genome and (iii) are located in said E gene region;(b) providing a second vector comprising an insertion nucleic acid flanked by said first and third restriction endonuclease sites;(c) contacting each of said first and second vectors with said first and third restriction endonucleases to produce first and second restriction endonuclease cleavage products, wherein said first cleavage product comprises said adenoviral genome and said second cleavage product comprises said insertion nucleic acid;(d) combining said first and second cleavage products under ligation conditions sufficient to produce a ligation product composition comprising a ligation product that includes said recombinant adenoviral genome having an insertion nucleic acid located in an E gene region of said genome;whereby said recombinant adenoviral genome having an insertion nucleic acid located in an E gene region of said genome is produced.