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

doi:10.1016/0006-291X(88)90636-5

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 journal › Article › peer-review

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 language	English (US)
Pages (from-to)	598-607
Number of pages	10
Journal	Biochemical and Biophysical Research Communications
Volume	151
Issue number	1
DOIs	https://doi.org/10.1016/0006-291X(88)90636-5
State	Published - Feb 29 1988
Externally published	Yes

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

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10.1016/0006-291X(88)90636-5

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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.",

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AU - Lee, S. G.

AU - Ricca, George A.

AU - Crumley, Gregg

AU - Lloyd, R. Stephen

AU - Drohan, William

PY - 1988/2/29

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N2 - 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.

AB - 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.

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

Abstract

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