摘要
目的克隆、表达发热伴血小板减少综合征布尼亚病毒(SFTSV)非结构蛋白NSs,对其功能初步鉴定。方法采用PCR法扩增经密码子优化后的SFTSV NSs基因,并克隆至PGEX-4T-2载体中,构建原核表达载体PGEX-4T-2-NSs,经酶切、测序鉴定后转化表达菌BL21(DE3),再经诱导、纯化得到谷胱甘肽S-转移酶(GST)-NSs融合蛋白,用Western Blot验证融合蛋白GST-NSs的抗原性。结果成功构建了GST-NSs融合蛋白原核表达载体,并在BL21细菌中获得高效表达;纯化后的融合蛋白具有良好的抗原性。结论实现了SFTSV重组NSs蛋白的原核高效表达,为进一步深入研究其结构与功能奠定了基础。
Objective To clone and express non-structural protein（NSs）of severe fever with thrombocytopenia syndrome bunyavirus （SFTSV）;to characterize preliminary function of recombinant protein.Methods SFTSV NSs coding region was optimized and amplified by PCR,which was cloned into PGEX-4T-2 vector and validated by restriction digestion and sequence analysis.Confirmed expression vector was transformed into E.coli host strain BL21（DE3）,which was cultured and induced for expression of glutathione S-transferase（GST）-NSs recombinant protein .GST-NSs recombinant protein was purified and tested for its antigenicity by Western Blot analysis.Results Prokayotic GST-NSs expression vector was constructed successfully. GST-NSs recombinant protein was expressed very effectively in BL21 bacteria cultures.Purified recombinant GST-NSs main-tained good antigenicity.Conclusion SFTSV NSs recombinant protein was expressed in prokaryotic system with high efficien-cy ,which laid a solid foundation for further study of its structure and functions .
出处
《江苏预防医学》
CAS
2015年第3期12-15,共4页
Jiangsu Journal of Preventive Medicine
基金
国家科技重大专项(2013ZX09102029)
江苏医学重点人才基金项目(RC2011082)
江苏省科技支撑项目(BE2012768)
