High-level expression of human TFF3 in Escherichia coli |
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Authors: | Wang Haibo Tong Yuanpeng Fang Ming Ru Binggen |
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Institution: | Department of Biochemistry and Molecular Biology, National Laboratory and Protein Engineering, College of Life Science, Peking University, Beijing 100871, PR China. |
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Abstract: | 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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