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 foreign gene for gene transfer. Two kinds of replication defective human recombinant adenovirus vectors were used in this study, the first containing beta-galactosidase reporter gene (AdCMVLac-Z) and the second carrying a gene for porcine leptin gene (AdCMVpLeptin). AdCMVLac-Z was tested for its ability to transfer DNA into pig kidney and pituitary cells. These cells expressed Lac-Z transiently 48 hours after the infection. In addition, when the pig kidney cells expressing the Lac-Z were replated with low density for the formation of colonies from each cell, colonies of blue cells expressing Lac-Z were observed. These results demonstrate that human recombinant adenovirus can be used as a transducing viral vector for inducing long-term expression in pig kidney cells. We also constructed a recombinant adenovirus (AdCMVpLeptin) 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 AdCMVpLeptin 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 expression in pig cells.
Lee
K
K
Department of Animal and Dairy Science, University of Georgia, Athens 30602, USA.
Barb
C R
CR
Kraeling
R R
RR
Riley
R T
RT
Hartzell
D L
DL
McGraw
R A
RA
Azain
M J
MJ
Dean
R G
RG
Baile
C A
CA
D000256:Adenoviridae; D000818:Animals; D002478:Cells, Cultured; D004274:DNA, Recombinant / Q000378:metabolism; D004279:DNA, Viral / Q000378:metabolism; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D006801:Humans; D020738:Leptin / Q000096:biosynthesis / Q000235:genetics*; D013552:Swine; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics*
D004274:DNA, Recombinant; D004279:DNA, Viral; D020738:Leptin; D001616:beta-Galactosidase