【摘要】 为了筛选反应原性较好的猪流行性腹泻病毒(PEDV)S基因片段,利用生物学软件对PEDV S基因进行了抗原位点分析,在不破坏线性抗原位点的前提下将S基因分为连续的4段进行表达,Western-blot检测表明,S1、Ps420、S2、S3蛋白都有很好的抗原性。纯化这4种蛋白,并免疫家兔,分别制备了S1、Ps420、S2、S3抗血清,间接ELISA检测结果显示,这4种抗血清都能识别细胞培养的PEDV,其中S1抗血清的反应性最好,其P/N值达到8.01,其次为Ps420抗血清。免疫荧光试验表明,S1、Ps420抗血清能识别天然的PEDV病毒粒子。提示,S1、Ps420抗血清在病原检测诊断中有一定的潜在应用价值。更多还原

【Abstract】 To screen S gene segments encoding protein with good reactivity of porcine epidemic diarrhea virus(PEDV),the S gene was analyzed on antigenic site by bioinformatics softwares.The S gene was segmented into four overlapping sections,named S1,Ps420,S2 and S3 gene,then the four sections were cloned into the prokaryotic expression vector pGEX-6P-1 to generate recombinant plasmids pGEX-6P-S1,pGEX-6P-Ps420,pGEX-6P-S3,and pProHTa-S2,respectively.The four plasmids were transformed into Escherichia coli BL21(DE3)pLysS and induced with IPTG,respectively.Western-blotting showed that expressed proteins had good antigenicity.To detect the reactivity of the four purified proteins,they were utilized to immune rabbits to produce anti-serum against single factor.The indirect ELISA results demonstrated that the four anti-sera were able to recognize the natural virus propagated in tissue culture.Of these,S1-produced anti-serum was the best,and the P/N value reached 8.01.The immunofluorescence assay showed that the anti-sera produced by protein S1 and Ps420 could also identify natural PEDV particle.These results showed that the anti-sera produced by proteins S1 and Ps420 were potentially useful as diagnostic antibody against PEDV.更多还原