High-level expression of human interferon gamma in Escherichia coli under control of the pL promoter of bacteriophage lambda

Gene. 1984 Apr;28(1):55-64. doi: 10.1016/0378-1119(84)90087-8.

Abstract

Several recombinant plasmids have been constructed which direct high-level synthesis of mature human interferon gamma (IFN-gamma) in Escherichia coli using the inducible leftward promoter pL of phage lambda followed by a translational initiator region derived either from the phage MS2 replicase gene or the E. coli tryptophan attenuator region. Under these conditions, IFN levels of up to 25% of the total cellular protein can be achieved. The highest levels were obtained when a terminator of transcription was cloned downstream from the IFN-gamma sequence. IFN-gamma was almost entirely found in the initial pellet fraction and not in soluble extracts. Co-induction of the lysis genes derived from phage MS2 or from phage lambda, inserted downstream from the IFN-gamma sequence, did not enhance the biological activity present in the supernatant fraction.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacteriophage lambda / genetics*
  • Cloning, Molecular
  • DNA, Recombinant / metabolism
  • Escherichia coli / metabolism*
  • Gene Expression Regulation
  • Humans
  • Interferon-gamma / biosynthesis*
  • Interferon-gamma / genetics
  • Operon
  • Peptide Chain Initiation, Translational

Substances

  • DNA, Recombinant
  • Interferon-gamma