Cloning , Expression of Human sTNFR1 Gene and the Biological Activity of Its Recombinant Protein
- VernacularTitle:人sTNFR1基因的克隆、融合表达与生物学活性
- Author:
Lei FU
;
Shi-Fang PENG
;
De-Ming TAN
;
Hong-Bo LIU
;
- Publication Type:Journal Article
- Keywords:
Human sTNFR1 Clone Prokaryotic expression Immunine activity Biological activity
- From:
China Biotechnology
2006;0(07):-
- CountryChina
- Language:Chinese
-
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 TNF?on 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.
