High-level expression of human TFF3 in Escherichia coli

Department of Biochemistry and Molecular Biology, National Laboratory and Protein Engineering, College of Life Science, Peking University, Beijing 100871, PR China.
Peptides (Impact Factor: 2.62). 08/2005; 26(7):1213-8. DOI: 10.1016/j.peptides.2005.04.012
Source: PubMed


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.

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