Production of recombinant human pancreatic secretory trypsin inhibitor by Escherichia coli.
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.
Kikuchi
N
N
Shionogi Research Laboratories, Shionogi & Co., Ltd., Osaka.
Nagata
K
K
Horii
T
T
Miyazaki
S
S
Shin
M
M
Takimoto
N
N
Tsuruta
Y
Y
Tamaki
M
M
Teraoka
H
H
Yoshida
N
N
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*
D000596:Amino Acids; D009841:Oligonucleotides; D011994:Recombinant Proteins; D014361:Trypsin Inhibitors; D014359:Trypsin Inhibitor, Kazal Pancreatic