多杀性巴氏杆菌IbeB重组蛋白诱导小鼠抗攻击感染的免疫保护力

发布时间：2019-06-16 12:52

【摘要】：为了研究猪多杀性巴氏杆菌IbeB重组蛋白(rIbeB)的免疫保护性,根据GenBank中登录的多杀性巴氏杆菌ibeB基因序列设计1对引物扩增ibeB基因,并重组至pET-28a(+)构建重组表达质粒pET-ibeB,转化到大肠杆菌BL21(D E3),经IPTG诱导获得rIbeB。生物信息学分析表明,猪多杀性巴氏杆菌ibeB基因长1 392 bp,编码蛋白质含463个氨基酸。该蛋白质的理论分子质量为51.98 ku,具有2个外排蛋白结构域,属外膜外排蛋白超家族。蛋白序列比对发现,不同血清型的多杀性巴氏杆菌IbeB的同源性达99%以上。rIbeB诱导小鼠的免疫保护结果表明,第3次免疫后第14天小鼠血清中IgG抗体水平显著升高,明显高于佐剂对照组。攻毒后,对照组小鼠第3天的存活率为20%,7 d后全部死亡;免疫组第3天的存活率达70%,第7天的存活率为40%。结果表明,rIbeB蛋白具有很好的免疫原性,可诱导小鼠产生部分免疫保护力。
[Abstract]:In order to study the immune protection of recombinant protein (rIbeB) of Pasteurella multocida IbeB, a pair of primers were designed to amplify ibeB gene according to the sequence of ibeB gene of Pasteurella multocida registered in GenBank, and the recombinant expression plasmid pET-ibeB, was constructed and transformed into E. coli BL21 (D E3). RIbeB. was induced by IPTG. Bioinformatics analysis showed that the 1392 bp, encoded protein of Pasteurella multocida ibeB gene contained 463amino acids. The theoretical molecular weight of the protein is 51.98 ku, which has two exocrine protein domains and belongs to the outer membrane exocrine superfamily. Protein sequence alignment showed that the homology of different serotypes of Pasteurella multocida IbeB was more than 99%. The immune protection results of mice induced by rIbeB showed that the level of IgG antibody in serum of mice increased significantly on the 14th day after the third immunization, which was significantly higher than that of the adjuvant control group. After challenge, the survival rate of the control group was 20% on the 3rd day and all died on the 7th day, and the survival rate of the immunized group was 70% on the 3rd day and 40% on the 7th day. The results showed that rIbeB protein had good immunogenicity and could induce partial immune protection in mice.
【作者单位】：湖南农业大学动物科学技术学院;湖南农业大学动物医学院;
【基金】：湖南省自然科学基金资助项目(14JJ4077)
【分类号】：S852.612

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