发热伴血小板减少综合征布尼亚病毒非结构蛋白NSs原核表达载体的构建与表达  

Construction and expression of prokaryotic expression vector for non- structural protein NSs in severe fever with thrombocytopenia syndrome bunyavirus

作　　者：张文帅[1] 张黎[1] 迟莹[1] 温恬[1] 黄超[1] 焦永军[1] 

机构地区：[1]江苏省疾病预防控制中心,江苏南京210009

出　　处：《现代预防医学》2015年第10期1836-1838,1863,共4页Modern Preventive Medicine

摘　　要：目的利用大肠杆菌原核表达系统表达发热伴血小板减少综合征布尼亚病毒(SFTSV)NSs重组蛋白。方法采用RT-PCR技术,从SFTSV毒株提取的病毒总RNA中,扩增NSs全长基因,将其克隆至p ET22b(+)中构建原核表达载体p ET22b-NSs,经酶切及测序鉴定后转化表达菌BL21(DE3),IPTG诱导,通过Western Blot鉴定NSs蛋白的表达。结果酶切、测序证明重组原核表达载体p ET22b-NSs构建成功,免疫印迹法可见NSs基因编码蛋白表达。结论实现了发热伴血小板减少综合征布尼亚病毒重组NSs蛋白的原核高效表达,为进一步深入研究发热伴血小板减少综合征布尼亚病毒NSs蛋白的结构与功能奠定了基础。Objective The study utilized the E. coli prokaryotic expression system to express recombinant protein NSs of the severe fever with thrombocytopenia syndrome bunyavirus（SFTSV）. Methods Total RNA of virulent SFTSV strain was extracted and full-length NSs was amplified by RT-PCR and cloned into p ET22b（＋） to construct a prokaryotic expression vector p ET22b-NSs.After restriction digestion and sequencing, the vector was transformed into BL21（DE3）, followed by IPTG induction. The expression of NSs was determined by Western Blot. Results Restriction digestion and sequencing demonstrated the successful construction of the recombinant prokaryotic expression vector p ET22b-NSs, and the expression of NSs-encoding protein was visualized by Western Blot. Conclusion The efficient expression of recombinant SFTSV NSs has been achieved, providing the basis for the elucidation of the structure and function of SFTSV NSs protein.

关 键 词：发热伴血小板减少综合征布尼亚病毒 NSS 原核表达 免疫印迹 

分 类 号：R373.3[医药卫生—病原生物学]