[Construction and properties of the expression vector based on the temperature-regulated P'R promoter in phage lambda DNA]

Bioorg Khim. 1987 Nov;13(11):1561-9.
[Article in Russian]

Abstract

Plasmid expression vector using the temperature-regulated promoter P'R of bacteriophage lambda is described. The vector carries a combination of two regions of lambda cI857indgenome, that contain: 1) gene cI and promoter PR, and 2) gene Q and promoter P'R. Transcription or gene Q is initiated at promoter PR, which is controlled by cI857 repressor. The Q gene product acts as a positive regulator of RNA synthesis from P'R. At 37 degrees C, sufficient amounts of protein Q are synthesised to initiate the expression of the cloned gene from P'R. Inactivation of Q gene (by elimination of a single NcoI site) results in the loss of P'R expression activity in the vector. E. coli beta-galactosidase gene (lacZ) and human leukocyte interferon alpha 2 gene (ifn alpha 2) were cloned into a single EcoRI cleavage site under the control of P'R. These constructs express high levels of beta-galactosidase and interferon alpha 2 in E. coli at 37 degrees C.

Publication types

  • English Abstract

MeSH terms

  • Bacteriophage lambda / genetics*
  • Cloning, Molecular
  • DNA, Viral / genetics*
  • Gene Expression Regulation
  • Genes, Synthetic
  • Genetic Vectors*
  • Interferon Type I / genetics
  • Plasmids*
  • Promoter Regions, Genetic*

Substances

  • DNA, Viral
  • Interferon Type I