Improved plasmid vectors with a thermoinducible expression and temperature-regulated runaway replication

Gene. 1983 Apr;22(1):103-13. doi: 10.1016/0378-1119(83)90069-0.

Abstract

Improved expression vectors have been constructed which are derived from runaway-replication mutants of plasmid R1 and carry the strong leftward promoter (pL) of bacteriophage lambda. The activity of this promoter is controlled by a temperature-sensitive repressor, product of the phage gene cI cloned on a compatible plasmid. Heat induction leads to amplification of the plasmid copy number and at the same time turns on the promoter. At a short distance downstream from the promoter, unique EcoRI, BamHI, XbaI and HindIII sites are present. This system was used for high level expression of the T4 DNA-ligase gene; 3 h after induction the ligase amounted to about 20% of total cellular protein.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacteriophage lambda / genetics*
  • Escherichia coli / genetics*
  • Gene Amplification
  • Genetic Vectors*
  • Mutation
  • Operon
  • Plasmids*
  • Replicon
  • Species Specificity
  • Temperature