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PMID: 6234201 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

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

Gene ·Vol. 28 ·No. 1 ·1984-04-00 ·Pages 55-64

Simons G, Remaut E, Allet B, Devos R, Fiers W

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.

MeSH Terms
Bacteriophage lambda/genetics Cloning, Molecular DNA, Recombinant/metabolism Escherichia coli/metabolism Gene Expression Regulation Humans Interferon-gamma/biosynthesis,genetics Operon Peptide Chain Initiation, Translational
Chemicals
DNA, Recombinant Interferon-gamma
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Simons G
Remaut E
Allet B
Devos R
Fiers W
Article Info
Journal
Gene
Abbr.
Gene
ISSN
0378-1119
Published
1984-04-00
Pages
55-64
Language
English
Region
Netherlands
NLM ID
7706761
Subset
IM
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