A plasmid vehicle suitable for the molecular cloning and characterization of mammalian promoters.
We have constructed a plasmid, pPSG4, that carries only the coding sequences for polyomavirus (Py) small, middle and truncated large T-antigens. A unique HindIII site allows the introduction of foreign promoters directly in front of viral coding sequences. The simian virus 40 (SV40) early and late, adenovirus-2 (Ad-2) major late and herpes simplex virus-1 (HSV-1) thymidine kinase (TK) promoters all confer on pPSG4 the ability to transform rat embryonic fibroblasts with high efficiency. Sequential deletion of the 72 bp repeats, the 21 bp repeats and the TATA box from the SV40 early region in pPSG4 produced a 50, then 30 and then a further 5 to 10-fold decrease in transformation efficiency, respectively. Thus pPSG4 is a convenient vector for the cloning and characterization of mammalian promoters.
D000818:Animals; D000956:Antigens, Viral / Q000235:genetics; D001483:Base Sequence; D002460:Cell Line; D002471:Cell Transformation, Neoplastic; D002872:Chromosome Deletion; D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes; D004274:DNA, Recombinant / Q000378:metabolism; D005822:Genetic Vectors; D009154:Mutation; D009876:Operon; D010957:Plasmids; D011120:Polyomavirus / Q000235:genetics; D051381:Rats
D000956:Antigens, Viral; D004274:DNA, Recombinant; D004262:DNA Restriction Enzymes