Cloning , Expression of Human sTNFR1 Gene and the Biological Activity of Its Recombinant Protein

China Biotechnology

2007, Vol. 27

Issue (7): 88-93 DOI:

Cloning , Expression of Human sTNFR1 Gene and the Biological Activity of Its Recombinant Protein

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Abstract

Human sTNFR1 (soluble tumor necrosis factor receptor 1) gene was amplified by RT-PCR from Hela cells. A recombinant expression vector of sTNFR1-MBP was constructed in pMAL-c2x, and transformed into E. Coli JM109.It was sequenced and confirmed to be identifical to the sTNFR1 gene in data bank. Recombinant protein sTNFR1-MBP was induced by IPTG and purified by Amylose resin Affinity Chromatography. sTNFR1-MBP was binded to sTNFR1's antibody in Western-blotting. From MTT assays, the results showed that sTNFR1-MBP could effectively block the cytotoxicity mediated by TNFαon QSG7701 cells. Annexin V-FITC staining and flowcytometry were used to observe the recombinant protein's anti-apoptosis capacity and the recombinant protein has marked anti-apoptosis effect in vitro.sTNFR1-MBP had good biological activity and it will be employed in further study.

Key words： Human sTNFR1 Clone Prokaryotic expression Immunine activity Biological activity

Received: 30 January 2007 Published: 25 July 2007

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Cite this article:

. Cloning , Expression of Human sTNFR1 Gene and the Biological Activity of Its Recombinant Protein. China Biotechnology, 2007, 27(7): 88-93.

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https://manu60.magtech.com.cn/biotech/ OR https://manu60.magtech.com.cn/biotech/Y2007/V27/I7/88

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