新型鸭呼肠孤病毒σC蛋白的原核表达及其多克隆抗体制备

Prokaryotic expression and polyclonal antibody preparation of σC protein of novel duck reovirus

为了解新型鸭呼肠孤病毒(NDRV TH11株)σC蛋白的抗原性,采用RT-PCR方法扩增了NDRV TH11株σC蛋白的编码基因,将其克隆于原核表达载体pET-30a(+)中,转化大肠杆菌BL21(DE3),经IPTG诱导表达His-σC重组蛋白。SDS-PAGE和Western blot分析表明该重组蛋白获得高效表达,并具有较好的反应原性。以纯化的σC蛋白免疫实验兔制备了抗σC蛋白的多克隆抗体,间接ELISA检测结果显示其效价达1∶20 000以上;间接免疫荧光试验表明,多克隆抗体能够特异性识别NDRV的σC蛋白,显示出σC蛋白具有良好的免疫原性。

To investigate the immunogenicity of σC protein of a novel duck reovirus （ NDRV TH11 ） , the σC gene was cloned into pET-30a（ ＋） and expressed in E.coli BL21（DE3）.SDS-PAGE and Western blot analysis showed that the expressed had good immunological activity .The polyclonal antibody was obtained from the rabbit immunized with purified recombinant protein and its titer was about 1∶20 000 by detection of indirect ELISA .Indirect immunofluorescence assay also demonstrated that the polyclonal antibody reacted specially with the native NDRV σC protein.This study will be helpful for preparation of peptide vaccines and σC function study.