中国动物传染病学报Issue(6)：6-11,6.
猪δ冠状病毒Nsp5的体外表达及其蛋白酶活性分析
PROKARYOTIC EXPRESSION AND PROTEASE ACTIVITY OF PORCINE DELTACORONAVIRUS NSP5
摘要
Abstract
Porcine deltacoronavirus (PDCoV) is a newly identiifed enteropathogenic coronavirus, causing diarrhea in suckling piglets. The nonstructural protein 5 (Nsp5) encoded by coronavirus is a 3C-like protease, and plays a key role in the process of viral polyproteins and immune regulation. In this study, the Nsp5 gene of PDCoV CHN-HN-2014 strain was amplified and subcloned into prokaryotic expression vector pET-28a(+), generating the recombinant expressing plasmid. After induction with IPTG, the fusion protein Nsp5-His was efifciently expressed in E.coli BL21 (DE3) in soluble form. The recombinant protein was further puriifed with Ni-NTA afifnity column and its protease activity was detected by fluorescence resonance energy transfer (FRET) method. The results showed that the puriifed recombinant protein efifciently cut the chemosynthetic 12 peptide that contained the N-terminal cleavage site of PDCoV Nsp5, indicating that Nsp5 possessed protease activity. The progress in cloning, expression and analysis of protease activity of Nsp5 will lay the foundation for the future functional research of PDCoV.关键词
猪δ冠状病毒/Nsp5/蛋白酶活性Key words
Porcine deltacoronavirus/Nsp5/protease activity分类
农业科技引用本文复制引用
张欢,肖文婕,曾哲,朱心宇,董楠,彭贵青..猪δ冠状病毒Nsp5的体外表达及其蛋白酶活性分析[J].中国动物传染病学报,2016,(6):6-11,6.基金项目
国家科技支撑计划 （）
