Expression, purification and activity verification of human norovirus NS6 protein
10.3760/cma.j.issn.1003-9279.2019.06.018
- VernacularTitle: 人诺如病毒NS6蛋白的表达、纯化及活性验证
- Author:
Yue YUAN
1
;
Yan XIN
1
;
Lili PANG
2
;
Zhaojun DUAN
2
Author Information
1. Department of Immunology, Basic School of Medical Science, Inner Mongolia Medical University, Hohhot 010110, China
2. NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China
- Publication Type:Journal Article
- Keywords:
Human norovirus;
Nonstructural protein NS6;
Affinity purification;
Gel filtration chromatography;
Enzymatic digestion
- From:
Chinese Journal of Experimental and Clinical Virology
2019;33(6):646-649
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To express the NS6 nonstructural protein of human norovirus (NoV) in Escherichia coli, and to detect its enzymatic activity after purification.
Methods:Human NoV NS6 gene was cloned into the prokaryotic expression vector pDE1 and then was transformed into E. coli BL21 for expression. NS6 protein was purified by Ni-NTA affinity chromatography and Superdex 200 pg column. The activity of NS6 protein was determined by digestion of fusion protein 15VP1-6P at 37 ℃.
Results:Human NoV NS6 protein was stably and highly expressed in E. coli. After purification, the expressed product reached a purity of more than 95%, and the relative molecular weight of NS6 protein was about 23×103 Da. NS6 protein could cleave the fusion protein containing rhinovirus 3C cleavage site.
Conclusions:The nonstructural protein NS6 of human NoV was successfully expressed in Escherichia coli, which laid a foundation for further study on the pathogenesis of human NoV.
