Modulation of expression of the human gamma interferon gene in E. coli by site-directed mutagenesis

S. G. Lee, George A. Ricca, Gregg Crumley, Robert (Stephen) Lloyd, William Drohan

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Plasmids expressing 2 forms of human immune interferon (IFN-γ) in E. coli have been constructed: 1) pIFNTacI which expresses IFN-γ with an N-terminal amino acid sequence of met-cys-tyr-cys-gln-, and 2) pIFNTacII which is a derivative of pIFNTacI from which the 9 base pairs (bp) coding for the cys-tyr-cys have been deleted. Quantitation of Western blots showed that approximately 10-fold more IFN-γ was produced in cells harboring pIFNTacII (7.5% of total cellular protein) as compared to pIFNTacI. The IFN-γ expressed in E. coli pIFNTacII is biologically active and routinely recoverable at 109 units per liter. When examined microscopically, IPTG induced E. coli harboring either plasmid construction contains prominent cytoplasmic inclusion bodies.

Original languageEnglish (US)
Pages (from-to)598-607
Number of pages10
JournalBiochemical and Biophysical Research Communications
Volume151
Issue number1
DOIs
StatePublished - Feb 29 1988
Externally publishedYes

Fingerprint

Mutagenesis
Site-Directed Mutagenesis
Escherichia coli
Interferons
Genes
Inclusion Bodies
Modulation
Plasmids
Isopropyl Thiogalactoside
Base Pairing
Interferon-gamma
Amino Acid Sequence
Western Blotting
Derivatives
Amino Acids
human IFNG protein
Proteins

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Modulation of expression of the human gamma interferon gene in E. coli by site-directed mutagenesis. / Lee, S. G.; Ricca, George A.; Crumley, Gregg; Lloyd, Robert (Stephen); Drohan, William.

In: Biochemical and Biophysical Research Communications, Vol. 151, No. 1, 29.02.1988, p. 598-607.

Research output: Contribution to journalArticle

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