为研究猪传染性胃肠炎病毒(TGEV)纤突蛋白(S)上主要抗原位点的免疫原性,本研究将S基因上A、B、C、D四个位点进行RT-PCR扩增,构建重组质粒pET30a-S,诱导表达后Western-blot检测,表明目的蛋白具有良好的抗原性.将纯化的目的蛋白免疫小鼠3次,在初次免疫后第0,14,28,42天采血,并用间接ELISA方法监测血清抗体动态水平变化.结果表明,获得的重组蛋白能诱导免疫小鼠产生特异性的体液免疫,说明该蛋白具有发展成TGEV亚单位疫苗的潜力.
In order to identify the innnunogenicity of the major antigenic sites on spike protein (S) of transmissible gastroenteritis virus (TGEV), we amplifed four antigenic sites on S gene named A, B, C and D using RT-PCR, and then constructed recombinant plasmid pET30a-S to express the interested proteins .Western-blot analysis showed that recombinant protein had good immunogeniciiv. M ice were immunized with the purl fled recombinant protein for 3 times. Serum samples were collected on 0, 14,28 and 42 days affer the primary immunization, and then the serum antibody level was determined by ELISA. Our results showed that the recombinant protein couht induce immunized mice to produce specific humoral immune responses, indieating that the protein has the potential to develop a TGEV subunit vaccine.