Modulation of expression of the human gamma interferon gene in E. coli by site-directed mutagenesis.
Plasmids expressing 2 forms of human immune interferon (IFN-gamma) in E. coli have been constructed: 1) pIFNTacI which expresses IFN-gamma 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-gamma was produced in cells harboring pIFNTacII (7.5% of total cellular protein) as compared to pIFNTacI. The IFN-gamma 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.
Lee
S G
SG
Meloy Laboratories, Inc., Rorer Biotechnology, Inc., Springfield, Virginia 22151.
Ricca
G A
GA
Crumley
G
G
Lloyd
R S
RS
Drohan
W
W
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism / Q000648:ultrastructure; D005786:Gene Expression Regulation; D005798:Genes, Bacterial; D006801:Humans; D007118:Immunoassay; D002479:Inclusion Bodies / Q000032:analysis; D007371:Interferon-gamma / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics*; D009154:Mutation; D010957:Plasmids
D007371:Interferon-gamma