Overproduction of the toxic protein, bovine pancreatic DNaseI, in Escherichia coli using a tightly controlled T7-promoter-based vector

Gene. 1993 Dec 22;136(1-2):337-40. doi: 10.1016/0378-1119(93)90491-k.

Abstract

A synthetic gene coding for bovine pancreatic DNaseI has been cloned under the control of a T7 promoter present on the plasmid pET11. This construct yields a stable Escherichia coli transformant only when transcription from this promoter is tightly controlled. Production of recombinant DNaseI (reDNaseI) is achieved by infection of the cells with a mutant lambda phage, CE6, which carries the gene encoding T7 RNA polymerase. Induced bacterial cultures yield in excess of 2 mg per litre of reDNaseI after purification.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacteriophage T7 / genetics
  • Cattle
  • Cloning, Molecular
  • DNA-Directed RNA Polymerases / genetics
  • Deoxyribonuclease I / genetics*
  • Escherichia coli / genetics*
  • Genes, Synthetic
  • Pancreas / enzymology
  • Plasmids*
  • Promoter Regions, Genetic*
  • Viral Proteins

Substances

  • Viral Proteins
  • bacteriophage T7 RNA polymerase
  • DNA-Directed RNA Polymerases
  • Deoxyribonuclease I