High-level expression of human TFF3 in Escherichia coli.
A strategy for expression and purification of recombinant N-terminal human trefoil factor family-domain peptide 3 (hTFF3) in Escherichia coli was established. The gene of hTFF3 was synthesized to substitute the low-usage condons with corresponding high-usage synonymous condons. At the same time, the signal peptide of DsbC was added to the N-terminus of the hTFF3 gene. The mature recombinant hTFF3 was located in the periplasm of E. coli, which can be released by sonication. The protein was further purified by a two-step cation exchange chromatography mentod. The yield is about 14-15 mg/l of culture. The biological activity of purified hTFF3 was analyzed by cell-based apoptosis assay, which shows that the recombinant hTFF3 is biologically active.
Wang
Haibo
H
Department of Biochemistry and Molecular Biology, National Laboratory and Protein Engineering, College of Life Science, Peking University, Beijing 100871, PR China.
Tong
Yuanpeng
Y
Fang
Ming
M
Ru
Binggen
B
D001483:Base Sequence; D003062:Codon / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D006801:Humans; D007218:Industrial Microbiology; D008969:Molecular Sequence Data; D009077:Mucins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D009124:Muscle Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D010455:Peptides; D019897:Periplasm / Q000737:chemistry / Q000378:metabolism; D015202:Protein Engineering; D021382:Protein Sorting Signals / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D000071165:Trefoil Factor-3
D003062:Codon; D009077:Mucins; D009124:Muscle Proteins; D010455:Peptides; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; C082495:TFF3 protein, human; D000071165:Trefoil Factor-3