Article

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: 2.43). 08/2005; 26(7):1213-8. DOI:10.1016/j.peptides.2005.04.012 pp.1213-8
Source: PubMed

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.

0 0
 · 
0 Bookmarks
 · 
19 Views
Data provided are for informational purposes only. Although carefully collected, accuracy cannot be guaranteed. The impact factor represents a rough estimation of the journal's impact factor and does not reflect the actual current impact factor. Publisher conditions are provided by RoMEO. Differing provisions from the publisher's actual policy or licence agreement may be applicable.

Keywords

biological activity
 
cell-based apoptosis assay
 
corresponding high-usage synonymous condons
 
Escherichia coli
 
hTFF3
 
hTFF3 gene
 
mature recombinant hTFF3
 
purification
 
purified hTFF3
 
recombinant hTFF3
 
recombinant N-terminal human trefoil factor family-domain peptide 3
 
sonication
 
two-step cation exchange chromatography mentod
 

Haibo Wang