A BK virus episomal vector for constitutive high expression of exogenous cDNAs in human cells.
A BK virus (BKV) episomal vector (pRPneoCMV) was constructed for expression of cDNAs under control of the cytomegalovirus (CMV) immediate-early promoter. Transfection of pRPneoCMV for expression of the chloramphenicol acetyltransferase (CAT) gene in several human cell lines showed that the CMV promoter is more efficient than the HIV-1 and RSV LTRs in directing gene expression from episomal vectors. In 293 human cells pRPneoCMV/CAT is twenty times more active in CAT expression than the well known pSV2CAT vector in COS7 cells. Stable expression of the gene of the herpes simplex virus type 1 and type 2 glycoprotein G, cloned into pRPneoCMV, was obtained in 293 cells. This vector will allow direct cloning of newly synthesized cDNAs whose expression can be monitored in human cells.
Sabbioni
S
S
Institute of Microbiology, School of Medicine, University of Ferrara, Italy.
Negrini
M
M
Rimessi
P
P
Manservigi
R
R
Barbanti-Brodano
G
G
D001739:BK Virus / Q000235:genetics*; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D003001:Cloning, Molecular / Q000379:methods*; D003587:Cytomegalovirus / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics*; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D010957:Plasmids; D011401:Promoter Regions, Genetic; D014162:Transfection; D014759:Viral Envelope Proteins / Q000235:genetics
D018076:DNA, Complementary; D014759:Viral Envelope Proteins; C045464:glycoprotein G, herpes simplex virus type 2; C050591:glycoprotein gG-1, herpes simplex virus type 1; D015500:Chloramphenicol O-Acetyltransferase