Expression of beta-galactosidase and pig leptin gene in vitro by recombinant adenovirus

Adenovirus has been used in vivo and in vitro as a vector to carry a foreig n gene for gene transfer. Two kinds of replication defective human recombin ant adenovirus vectors were used in this study, the first containing beta-g alactosidase reporter gene (AdCMVLac-Z) and the second carrying a gene for porcine leptin gene (AdCMVpLeptin). AdCMVLac-Z was tested for its ability t o transfer DNA into pig kidney and pituitary cells. These cells expressed L ac-Z transiently 48 hours after the infection. In addition, when the pig ki dney cells expressing the Lac-Z were replated with low density for the form ation of colonies from each cell, colonies of blue cells expressing Lac-Z w ere observed. These results demonstrate that human recombinant adenovirus c an be used as a transducing viral vector for inducing long-term expression in pig kidney cells. We also constructed a recombinant adenovirus (AdCMVpLe ptin) which contained a pig leptin gene for the expression of pig leptin in vitro in the 293 human kidney cell line. 293 cells transfected with AdCMVp Leptin produced both a 15 KDa of a secretory form of porcine leptin and an 18 KDa long form containing signal peptide. Our study demonstrated that the recombinant adenovirus system offers a method for gene transfer and expres sion in pig cells.