METHOD FOR RAPIDLY PRODUCING RECOMBINANT VIRUSES FROM HOST CELLS USING SITE-SPECIFIC RECOMBINATION TECHNOLOGY WITHOUT FURTHER PROCEDURES

摘要

PURPOSE: A method for producing recombinant viruses using site-specific recombination technology is provided. Desired recombinant viruses are rapidly produced from host cells directly without further procedures such as selection of the desired recombinant viruses from intermediate host cells because un-bound viral genomic DNA segments cannot be expressed by themselves, but only viruses having desired DNA fragments can be expressed. CONSTITUTION: The method for preparing recombinant viruses having linear genomic DNAs comprises the steps of: (i) providing viral genomic DNA segments containing site-specific recombination sites and nucleotide sequences required for viral replication and packaging; (ii) constructing carrier vectors containing at least one desired DNAs and site-specific recombination sequences; (iii) cloning the genomic DNA segments of step (i) into the carrier vectors of step (ii) in vitro to produce a mixture comprising expressive linear recombinant viral vectors; and (iv) transfecting the mixture of step (iii) to host cells, wherein the virus is adenovirus, retrovirus, adeno-associated virus, bacteriophage, coronavirus and herpes simplex virus, CMV(cytomegalovirus), EBV(Epstein-Barr virus) or VZV(Varicella-zoster virus).