Expression,purification and cleavage activity analysis of self-processed recombinant MBP-HRV 3C fusion protease in E.coli expression system
10.3969/j.issn.1673-4130.2014.20.001
- VernacularTitle:自剪切重组麦芽糖结合蛋白-人鼻病毒3C 蛋白酶融合蛋白在大肠杆菌表达系统中表达、纯化及活性检测
- Author:
Zhejun DONG
;
Haijian ZHAO
;
Xiaomao XU
;
Baomin FANG
;
Jian GUO
;
Fei XIAO
- Publication Type:Journal Article
- Keywords:
HRV 3C protease;
expression;
tool-enzyme
- From:
International Journal of Laboratory Medicine
2014;(20):2721-2722,2725
- CountryChina
- Language:Chinese
-
Abstract:
Objective To obtain a novel tool-enzyme for genetic engineering with good solubility,strong specificity of enzyme digestion and maintaining the enzyme activity at low temperature by using E.coli expression system to express self-processed re-combinant MBP-HRV 3C fusion protease.Methods The cDNA encoding HRV 3C protease was cloned into pRSF-Duet vector.The recombinant plasmid was transferred into E.coli BL21 (DE3)for expression.HRV 3C protease was obtained through Nichol col-umn affinity purification.The cleavage activity of HRV 3C protease was determined by in vivo experiment.Results HRV 3C prote-ase was highly expressed in E.coli expression system,and the obtained HRV 3C protease could recognize and digest HRV 3C site. Conclusion A novel tool-enzyme for genetic engineering is obtained.
