title: Production of recombinant human pancreatic secretory trypsin inhibitor by Escherichia coli.
abstract: A synthetic gene for human pancreatic secretory trypsin inhibitor (PSTI) was fused to the coding sequence for the amino-terminal 135 amino acid residues of human interferon-gamma (IFN-gamma) by interposing a methionine codon sequence, and the resulting hybrid gene was efficiently expressed in Escherichia coli cells. Recombinant human PSTI (rHu-PSTI) was separated from the IFN-gamma/PSTI fused protein by cleavage at the methionine residue with cyanogen bromide. Finally, rHu-PSTI was purified by affinity chromatography on a bovine trypsin-CH-Sepharose 4B column. The amino acid composition, partial amino-terminal sequence, disulfide formation, human trypsin inhibitory activity, and immunoreactivity against rabbit anti-human PSTI serum of rHu-PSTI corresponded to those of the natural form.
authors.affiliation: Shionogi Research Laboratories, Shionogi & Co., Ltd., Osaka.
authors.forename: N
authors.initials: N
authors.lastname: Kikuchi
authors.forename: K
authors.initials: K
authors.lastname: Nagata
authors.forename: T
authors.initials: T
authors.lastname: Horii
authors.forename: S
authors.initials: S
authors.lastname: Miyazaki
authors.forename: M
authors.initials: M
authors.lastname: Shin
authors.forename: N
authors.initials: N
authors.lastname: Takimoto
authors.forename: Y
authors.initials: Y
authors.lastname: Tsuruta
authors.forename: M
authors.initials: M
authors.lastname: Tamaki
authors.forename: H
authors.initials: H
authors.lastname: Teraoka
authors.forename: N
authors.initials: N
authors.lastname: Yoshida
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D001483:Base Sequence; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005813:Genes, Synthetic; D006801:Humans; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000138:chemical synthesis; D011994:Recombinant Proteins / Q000096:biosynthesis*; D014359:Trypsin Inhibitor, Kazal Pancreatic / Q000096:biosynthesis*; D014361:Trypsin Inhibitors / Q000096:biosynthesis*
chemical_list: D000596:Amino Acids; D009841:Oligonucleotides; D011994:Recombinant Proteins; D014361:Trypsin Inhibitors; D014359:Trypsin Inhibitor, Kazal Pancreatic