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。
To explore immune protective efficacy induced by porcine Pasteurella multocidarIbeB, the primers of ibeBgene of P. multocifla were designed based on the ibeBgene sequence retrieved from GenBank. The ibeBgene was cloned by PCR and subcloned into pET-28a（＋） to construct recombinant plasmid pET-ibeB. The pET-ibeBwas then transformed into Escherichia coli BL21（DE3）.rIbeB was expressed after being induced by IPTG. The bioinformatics analysis results showed that the ibeB gene consisted of 1 392 bp, encoding 463amino acids of protein with a theoretical molecular weight of 51.98ku. The IbeB was an outer membrane protein with 2 outer membrane efflux protein domains, belonging to outer membrane efflux protein subfamily. The IbeB had above 99% homology between different serotypes of ~ multocida. The results on immune protective efficacy showed that serum IgG levels significantly increased in immuned group at 14 days after the third immunization with rIbeB and was significantly higher than that in adjuvant group. After being challenged with ~ multocida PM-C349 strain,the mice in the adjuvant group showed 20% survival rate at 3 days and all died at 7 days,but the mice in the immunized group showed 70% survival rate at 3 days and 40% survival rate at 7 days. In conclusion,the rIbeB has good immunogenicityand induces partial immune protective efficacy against infection with P.multocidaPM-C349 strain in mice.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2017年第5期630-635,共6页
Chinese Veterinary Science
基金
湖南省自然科学基金资助项目(14JJ4077)
