鹅坦布苏病毒囊膜蛋白结构域 II 在大肠杆菌中的表达及免疫学鉴定

摘要

为了实现鹅坦布苏病毒囊膜蛋白( E蛋白)结构域II在大肠杆菌中的表达并对其进行免疫学鉴定,根据鹅坦布苏病毒JS804株囊膜蛋白基因序列,利用人工合成法获得鹅坦布苏病毒囊膜蛋白结构域II编码基因并引入限制性酶切位点,片段大小为507 bp。将合成的基因片段克隆入pGEX-4t-1原核表达载体中,构建重组表达载体pGEX-EII并转化至BL21(DE3)中。通过优化表达条件获得重组蛋白质并进行免疫学鉴定。结果显示,重组蛋白质分子量约为4.4×104,Western-blot和间接免疫荧光检测结果表明重组蛋白质具有良好的免疫原性。%To express and immunoidentify goose tembusu virus envelope domain II in Escherichia coli, the encoding gene of goose tembusu virus JS804 strain envelope protein domain II was artificially synthesized with the length of 507 bp. The synthesized gene was then inserted into the prokaryotic vector pGEX-4t-1 for the construction of recombinant expression plasmid pGEX-EII and then transformed into E. coli BL21 ( DE3 ) . The recombinant protein with the molecular mass of 4. 4×104 was successfully expressed in E. coli by optimizing the conditions of expression. Western-blot and IFA revealed the good immunogenicity exhibited by the recombinant protein.