中文摘要：

为了实现鹅坦布苏病毒E蛋白质结构域I和II在大肠杆菌中的表达。利用PCR方法扩增得到目的片段，并引入FLAG标签序列（DYKDDDDK），构建重组表达质粒pET28a-E-I/II。转化至BL21（DE3）中，经IPTG诱导表达目的蛋白质，并对其进行SDS-PAGE分析和Western-blot鉴定。结果显示， PCR扩增得到约900 bp的目的片段，经鉴定成功构建重组表达载体pET28a-E-I/II。重组菌在IPTG诱导4 h后在37000处出现目的蛋白质并且表达量达到高峰。经超声波破碎后SDS-PAGE分析显示，融合蛋白质以包涵体形式存在。 Western-blot结果显示，重组蛋白质与FLAG单抗和E蛋白质阳性血清均可发生特异性反应。表明鹅坦布苏病毒E蛋白质结构域Ⅰ和Ⅱ在大肠杆菌中成功表达，经鉴定重组蛋白质具有良好的免疫反应性。

英文摘要：

Envelope protein domains I and Ⅱ gene of goose tembusu virus was amplified by PCR with the specific primers designed on the basis of E gene, and was inserted into the prokaryotic vector pET28a for the construction of recom-binant expression plasmid pET28a-E-I/II. pET28a-E-I/II was transformed into Escherichia coli BL21 （DE3）. The recom-binant protein was induced by IPTG and identified by SDS-PAGE and Western-blot. After 4 h induction by IPTG, the re-combinant protein with the molecular weight of 37 000 reached the yield peak. The SDS-PAGE analysis after sonication showed that the fusion protein was expressed in Escherichia coli as inclusion bodies. Western-blot revealed that the recombinant protein specifically reacted with FLAG McAb and positive serum for E protein. Goose tem-busu virus envelope domains I andⅡwas successfully ex-pressed in Escherichia coli and identified to be immune ac-tive.