Modulation of expression of the human gamma interferon gene in [formula omitted] by site-directed mutagenesis
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-ty...
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Published in | Biochemical and biophysical research communications Vol. 151; no. 1; pp. 598 - 607 |
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Main Authors | , , , , |
Format | Journal Article |
Language | English |
Published |
Elsevier Inc
1988
|
Online Access | Get full text |
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Summary: | 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 10
9 units per liter. When examined microscopically, IPTG induced
E.
coli
harboring either plasmid construction contains prominent cytoplasmic inclusion bodies. |
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ISSN: | 0006-291X 1090-2104 |
DOI: | 10.1016/0006-291X(88)90636-5 |