Dual-origin plasmids containing an amplifiable ColE1 ori; temperature-controlled expression of cloned genes.
Dual-origin plasmids comprising an inducible ColE1-derived origin of replication controlled by the lambda pR promoter, the cI857 temperature-sensitive repressor gene and the pSC101 origin of replication and its associated par sequence, were constructed. Such plasmids carrying cloned genes were stably maintained at four copies per chromosome, and were readily amplifiable by thermal induction. Cloned gene expression increased with copy number, and accumulation values of greater than 20% total cellular protein were detected. These vectors should prove useful for the production of foreign protein on a large scale, since they provide for stable plasmid maintenance during the growth phase, and high-level gene expression without plasmid loss during the production phase.
D000123:Acetyltransferases / Q000235:genetics; D000818:Animals; D003086:Bacteriocin Plasmids; D010582:Bacteriophage lambda / Q000235:genetics; D002417:Cattle; D015500:Chloramphenicol O-Acetyltransferase; D012085:Chymosin / Q000235:genetics; D003001:Cloning, Molecular / Q000379:methods; D004261:DNA Replication; D004792:Enzyme Precursors / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005784:Gene Amplification; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012097:Repressor Proteins / Q000235:genetics; D013696:Temperature; D014158:Transcription, Genetic
D004792:Enzyme Precursors; D012097:Repressor Proteins; D000123:Acetyltransferases; D015500:Chloramphenicol O-Acetyltransferase; C032443:prorennin; D012085:Chymosin