摘要
本文以IBV N蛋白为研究对象,首先从鸡胚培养的IBV ZY3毒株尿囊液中提取基因组RNA,然后利用软件对其N蛋白抗原表位进行预测分析,设计一对特异性引物对免疫原性好、亲水性强、表位可能性大的保守片段进行扩增,片段大小为818 bp;将扩增产物与表达载体pET-32a(+)连接并转入大肠杆菌BL21中诱导表达,用SDS-PAGE检测表达的重组N蛋白大小为50 kDa;用Ni-NTA Superflow纯化重组N蛋白后,采用抗鸡IBV血清与之进行Western blot反应,检测其免疫原性,结果显示重组蛋白免疫原性良好。该结果对进一步研究IBV N蛋白及建立鸡传染性支气管炎的诊断方法有重要意义。
IBV N protein was regarded as a target protein in our study, IBV genomic RNA was extracted from chicken embryo allantoic fluid, using software to predict and analyse the epitope of N protein. A pair of specific primers were designed to amplify the fragments which showed good immunogenic, hydrophilicity and epitope possibility, amplified fragment was 818 bp. It ligated into the vector pET-32a （＋） and then transformed into E. coli BL21, after induction of expression, the SDS-PAGE results showed that the protein was about 50 kDa. Purified on a column packed with Ni-NTA His＇Bind superflow, the recombinant protein then reacted with chicken anti-IBV serum by western blot to test its antigenicity, the results showed that the recombinant protein had good antigenicity. It was important to further research on IBV N protein and the diagnosis of avian infectious bronchitis.
出处
《四川畜牧兽医》
2017年第4期24-27,共4页
Sichuan Animal & Veterinary Sciences
