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| Expression and Characterization of a Novel Truncated TGF-&beta|Receptor II Synthesized in Escherichia coli |
| CHU Pan-Hui1,2**, LIU Hai-Feng1, WANG Xiao-Hua3, ZHANG Hu-Fei1, WU Yan1, YUAN Xiao-Huan1 |
1.Heilongjiang Key Laboratory of Antifibrosis Biotherapy, Mu Dan Jiang Medical University, Mudanjiang157011, China
2.College of Life Science and Technology, Jinan University, Guangzhou510632,China
3.The Laboratory of Pathogeny Biology, Mu Dan Jiang Medical University, Mudanjiang157011, China |
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Abstract To clone human truncated TGF-β receptor II（tTGF-βRⅡ）and express it in Escherichia coli. The gene tTGF-βRⅡ,obtained by PCR was cloned into the vector pGEX4T3 to construct a fusion expression plasmid.The recombinant protein GST-tTGF-βRⅡ were expressed in soluble form, purified by Glutathione-Sepharose 4B affinity chromatography and cleaved by thrombin to release tTGF-βRⅡ. Purification of tTGF-βRⅡ was achieved by affinity chromatography. SDS-PAGE indicated that fusion protein GST- tTGF-βRⅡ was secreted as a protein of around 37.0 KDa and aimed protein tTGF-βRⅡwas 11.0 KDa. The GST-tTGF-βRⅡand tTGF-βRⅡ protein were confirmed by Western blot analysis. The purification of aimed protein was biologically active in a validated inhibited human fibroblast proliferation bioassay.
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Received: 26 March 2009
Published: 28 July 2009
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