Patent ID: 6706477
Filing Date: 2004-03-16
Classification: C12N

Abstract:
A method of constructing a library comprising a plurality of heterologous nucleic acids, comprising:(a) cleaving an isolated vaccinia virus genome within the tk gene to produce a first viral fragment and a second viral fragment, wherein said first fragment is nonhomologous with said second fragment; (b) providing a population of transfer plasmids comprising said heterologous nucleic acids flanked by a 5â€² flanking region and a 3â€² flanking region, wherein said 5â€² flanking region is homologous to said first viral fragment and said 3â€² flanking region is homologous to said second viral fragment; and wherein said transfer plasmids are capable of homologous recombination with said first and second viral fragments such that a viable vaccinia virus genome is formed; and (c) introducing said transfer plasmids and said first and second viral fragments into a mammalian host cell under conditions wherein a transfer plasmid and said viral fragments undergo in vivo homologous recombination, thereby producing a library of viable modified vaccinia virus genomes comprising heterologous nucleic acids.