中文摘要：

【目的】明确鹅坦布苏病毒（GTUMV）E蛋白结构域I的原核表达及免疫原性,为进一步研究GTMUV E蛋白结构的生物学特性、免疫学功能等奠定基础。【方法】通过人工合成方法获得GTMUV E蛋白结构域I的编码基因（EI基因）,并与携带GST标签的p GEX-4t-1载体连接,转入大肠杆菌后经诱导表达获得融合蛋白,并以Western blotting鉴定融合蛋白是否有免疫原性。【结果】合成获得的E1基因片段为411 bp,将其插入p GEX-4t-1载体可构建重组表达质粒p GEX-4t-1-EI。阳性重组菌经1 mmol/L IPTG诱导5 h后,融合蛋白的表达量达最高峰,且以包涵体形式存在,分子量约41.0 k Da。Western blotting鉴定结果显示,融合蛋白与GST标签抗体和E蛋白阳性血清均可发生特异性反应,检测到预期的目的条带。【结论】GTMUV E蛋白结构域I可在大肠杆菌中成功诱导表达,且获得的融合蛋白具有良好的免疫原性,可用于GTMUV血清学检测试剂盒的研发。

英文摘要：

【Objective】The prokaryotic expression and immunogenicity of goose tembusu virus（GTMUV） E protein domain I were identified in order to lay the foundation for further researches on biological characteristics and immunological function of E protein domain I in GTMUV. 【Method】The encoding gene（EI） of GTMUV E protein domain I was artificially synthesised and was inserted into vector p GEX-4t-1 with GST tag. Then the recombinant plasmid was transformed into Escherichia coli. Through induced expression, the fusion protein was obtained and its immunogenicity was identified by using Western blotting. 【Result 】The obtained gene fragment E1 was 411 bp in length. Through inserting into vector p GEX-4t-1, the recombinant expression plasmid p GEX-4t-1-EI was constructed successfully. When positive recombinant strain was induced with 1 mmol/L IPTG, the expression quantity of fusion protein reached the peak after 5 hours. The fusion protein existed as inclusion body in E. coli, and the molecular weight was about 41.0 k D. The identification result of Western blotting showed that fusion protein could react specifically with GST Mc Ab and E protein positive serum to detect out target bands. 【Conclusion】The E protein domain I of GTMUV can be successfully expressed in E. coli, and the obtained fusion protein has good immunogenicity. Therefore it can be used for research development of GTMUV serology detection kit.