Expression, purification and activity assay of human IL-37b in E.coli
10.3969.j.issn.1671-7856.2017.03.004
- VernacularTitle:人IL-37b在大肠埃希菌中的表达、纯化及活性鉴定
- Author:
Mengyuan LI
;
Rongfei WEI
;
Damo XU
;
Xingjiu YANG
;
Ran GAO
- Keywords:
IL-37b;
Expression;
Purification;
Biological activity
- From:
Chinese Journal of Comparative Medicine
2017;27(3):20-24
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the expression of recombinant IL-37b protein and removal of the endotoxin, and identify its biological activity.Methods The prokaryotic expression vector pET28/IL-37b was constructed and to transform Escherichia coli (E.coli) Rosetta.After induction with IPTG, the recombinant protein was purified through Ni2+-NTA gel column and identified by SDS-PAGE and Coomassie brilliant blue staining.Then, the endotoxin protein was removed and was treated with LPS-stimulated RAW 264.7 cells.The culture supernatant was collected.The expression of IL-6 was detected by ELISA and the biological activity of the protein was identified.Results The recombinant IL-37b with high purity was expressed and the endotoxin produced by prokaryotic expression was reduced, and it was identified to have good biological activity.Conclusions In this study a recombinant IL-37b protein with high biological activity is successfully obtained.
