Establishment of gene therapy vector by using DNA replication origin and virus vector

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Expression vector composed of adenovirus, beta-galactosidase, and DNA replication origin (ori) was constricted.Oris used here were from genes of c-myc, human hsp70 and immunoglobulin heavy chain in addition to SV40 origin as a positive control. These origins were inserted to adenovirus vector containing Cre-lox system, and the recombinant adenovirus were recovered from human 2983 cells transfected with these vectors. Adenovirus containing SV40 origin were first infected to monkey CosI cells, and the circular adenovirus expressing beta-galactosidase was recovered. Adenovirus containing hsp70 gene origin were then infected to human HeLa cells, and the circular form of adenovirus DNA was also recovered. Copy number of hsp70 ori derived plasmid DNA in the cells, however, was lower than that of SV40 origin in 10^<-3> order. Results suggest that this system should be useful for an expression vector for gene therapy.