High throughput system for producing recombinant viruses using site-specific recombination

  • US 7,319,001 B2
  • Filed: 03/07/2003
  • Issued: 01/15/2008
  • Est. Priority Date: 03/09/2002
  • Status: Active Grant
First Claim
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1. A method for preparing expressive circular recombinant viral vectors, comprising:

  • i) providing a plurality of linearized viral DNA fragments wherein each fragment contains at least two site-specific recombination sites and at least one restriction enzyme recognition site;

    ii) providing a plurality of viral vector carriers wherein each viral vector carrier contains a gene cassette having a desired genomic material with a desired nucleic acid sequence flanked with at least two site-specific sites for homologous recombination with the linearized DNA fragments at the two site-specific recombination sites; and

    iii) reacting the linearized viral DNA fragments and the viral vector carriers to create homologous recombinations in vitro wherein a plurality of recombinant viral vectors carrying the desired genomic material is formed in vitro therefrom;

    iv) wherein non-recombined linearized viral DNA fragments, non-recombined viral vector carriers, and non-linearized viral DNA do not express a gene for a protein responsible for viral replication and viral particle formation.

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