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

• Published in: Gene Vol. 28; no. 1; pp. 55 - 64
• Main Authors: Simons, Guus, Remaut, Erik, Allet, Bernard, Devos, Rene, Fiers, Walter
• Format: Journal Article
• Language: English
• Published: Lausanne Elsevier B.V 01.04.1984; Amsterdam Elsevier; New York, NY
• Subjects: Antiviral drugs; Bacteriophage lambda - genetics; Biological and medical sciences; Biotechnology; Cloning, Molecular; DNA, Recombinant - metabolism; Escherichia coli - metabolism; Fundamental and applied biological sciences. Psychology; Gene Expression Regulation; Health. Pharmaceutical industry; Humans; Industrial applications and implications. Economical aspects; Interferon-gamma - biosynthesis; Interferon-gamma - genetics; lysis genes; Microbiology; Operon; pellet fraction; Peptide Chain Initiation, Translational; plasmid expression vectors; Production of active biomolecules; Recombinant DNA; Replicative cycle, interference, host-virus relations, pathogenicity, miscellaneous strains; transcription terminator; Virology; transcription terminator; IFN-γ; lysis genes; SDS; TCA; DTT; plasmid expression vectors; S.D; pellet fraction; Recombinant DNA; P L; bp; Human; Bactériophage lambda; Escherichia coli; Gene expression; Virus; Lambda phage group; Transcription promotor; Styloviridae; Bacteria; Bactériophage; Escherichieae; Vector; Enterobacteriaceae; Gamma interferon
• ISSN: 0378-1119; 1879-0038
• DOI: 10.1016/0378-1119(84)90087-8

Several recombinant plasmids have been constructed which direct high-level synthesis of mature human interferon gamma (IFN-γ) in Escherichia coli using the inducible leftward promoter P L of phage λ 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-γ sequence. IFN-γ 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 λ, inserted downstream from the IFN-γ sequence, did not enhance the biological activity present in the supernatant fraction.