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

S. G. Lee, George A. Ricca, Gregg Crumley, R. Stephen Lloyd, William Drohan

Research output: Contribution to journalArticle

3 Scopus citations

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

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ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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