New BK virus episomal vector for complementary DNA expression in human cells.
The properties of pRP-c, a new vector for complementary DNA (cDNA) expression, are described. The vector contains the early region and replication origin of BK virus (BKV), a human papovavirus. Due to the presence of these BKV sequences, pRP-c replicates in human cells allowing amplification of inserted cDNAs. The promoter, intron and polyadenylation region for cDNA expression are separated by unique restriction sites and can therefore be individually excised and substituted with different transcription signals. Coding sequences of the bacterial genes for chloramphenicol-acetyl transferase (CAT) or neomycin phosphotransferase (neo) were inserted into the cDNA cloning site of pRP-c and expressed in human cells in transient assays or stable clones. In both cases expression of the inserted sequences was significantly more efficient than by using the integration vectors pSV2CAT and pSV2neo, demonstrating the advantages of episomal expression vectors in human cells. Possible uses of pRP-c to express viral and cellular cDNAs in human cells are discussed.
Grossi
M P
MP
Institute of Microbiology, School of Medicine, University of Ferrara, Italy.
Caputo
A
A
Rimessi
P
P
Chiccoli
L
L
Balboni
P G
PG
Barbanti-Brodano
G
G
D001739:BK Virus / Q000235:genetics*; D015139:Blotting, Southern; D002478:Cells, Cultured; D015500:Chloramphenicol O-Acetyltransferase / Q000096:biosynthesis; D003001:Cloning, Molecular; D004247:DNA / Q000096:biosynthesis; D004262:DNA Restriction Enzymes / Q000494:pharmacology; D004279:DNA, Viral / Q000096:biosynthesis*; D005822:Genetic Vectors; D006801:Humans; D010957:Plasmids; D011120:Polyomavirus / Q000235:genetics*; D014162:Transfection
D004279:DNA, Viral; D004247:DNA; D015500:Chloramphenicol O-Acetyltransferase; D004262:DNA Restriction Enzymes