猪传染性胃肠炎病毒S蛋白抗原表位C的串联表达研究

摘要

为比较含有猪传染性胃肠炎病毒(TGEV)S蛋白抗原表位C不同表位密度的重组蛋白的抗原性,本研究将抗原表位C的编码核酸序列进行串联,构建了重组表达质粒pET-(C1C2)2～pET-(C1 C2)6,并在原核表达系统中表达了重组蛋白.Western blot分析表明,表达的6种重组蛋白r-CiC2～r-(CiC2)6均具有良好的抗原性.采用Dot-ELISA方法对各重组蛋白进行了抗原性比较分析.结果表明,含有12个表位肽的重组蛋白r-(CiC2)6的抗原性强于其它重组蛋白.本研究制备的串联重组蛋白为建立TGE血清学诊断方法奠定了物质基础.%To compare the antigenicity of transmissible gastroenteritis virus (TGEV) S recombinant protein with different epitopes density of antigenic epitope C,several recombinant plasmids from pET-(C1C2)2 to pET-(C1C2)6 were constructed including tandem epitope C,and these recombinant plasmids were expressed in E.coli Rosetta (DE3).All the six recombinant proteins of r-C1C2 to r-(C1C2)6 showed good antigenicity by westem blot analysis.Antigenicity analysis of the recombinant protein showed r-(C1C2)6 was more strong reactivity to TGEV-positive serum than that of the other recombinant proteins by Dot-ELISA.The tandem recombinant proteins prepared in this research laid the material foundation for establishing TGE serodiagnosis methods.