Expression, purification and activity verification of human norovirus NS6 protein / 中华实验和临床病毒学杂志
Chinese Journal of Experimental and Clinical Virology
; (6): 646-649, 2019.
Article
in Zh
| WPRIM
| ID: wpr-805393
Responsible library:
WPRO
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.
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Index:
WPRIM
Language:
Zh
Journal:
Chinese Journal of Experimental and Clinical Virology
Year:
2019
Type:
Article