鸭疫里默杆菌mazp基因克隆表达及免疫原性分析  

Cloning,Expression and Immunogenicity of mazp Gene from Riemerella Anatipestifer

作　　者：王秀祯 罗永芳[1] 夏文君[1] 罗雪刚 彭少静 周念[1] 李继祥[1] 

机构地区：[1]西南大学荣昌校区动物医学系,重庆402460

出　　处：《动物医学进展》2015年第2期44-47,共4页Progress In Veterinary Medicine

摘　　要：为探索鸭疫里默杆菌不同血清型菌株的共同保护性抗原,克隆表达mazp基因,并采用动物试验测定表达蛋白的免疫原性。根据RA-GD株mazp基因核苷酸序列设计特异性引物进行PCR扩增,克隆测序获得RA-AF株细菌1 326bp的核苷酸片段,比较分析发现其保守性高。以克隆的mazp基因构建重组表达质粒pET-32a-mazp并转化E.coli BL21(DE3),IPTG诱导获得重组蛋白。重组蛋白为免疫抗原免疫小鸭15d后,RA-BSY(血清1型)和RA-AF(血清2型)攻毒的保护指数分别为77.8和62.4;抗血清能延迟发病和死亡时间。结果表明,鸭疫里默杆菌mazp蛋白具有免疫原性,免疫鸭对血清1型和2型RA感染具有抵抗力。To study the mutual protective antigen of Riemerella anatipestifer （RA） strains, the reserch cloned and expressed membrane-associated zinc metalloprotease （mazp） gene of RA, and studied its expressed product immunogenicity using animal test. The specific primers were designed based on the nucleotide sequence of mazp gene of RA-GD strain and the mazp gene of RA-AF strain was cloned and sequenced. Compared with other strains through Blast analysis, the 1 326 bp nucleotide sequence of mazp gene of RA-AF strain was highly conserved. Then the mazp gene fragment was cloned into pET-32a （＋） to construct expression plasmid pET-32a-mazp. It expressed recombinant protein in E. coli BL21 （DE3） when induced by isopropyl thiogalactoside （IPTG）. Subcutaneous inoculation of the recombinant protein in 5-day-old ducklings gave adequate protective index （PI= 62.4） against challenge with RA-AF strain （serotype 2） 15 days after vaccination, while the protection （PI= 77.8） was obtained by challenge with RABSY strain （serotype 1）. The antiserum of recombinant protein was used as antibody to immunize ducklings, delaying morbility and death was obtained. The result indicated the recombinant protein mazp has inamunogenicity, and may play a role in protecting ducks from infection with Riemerella anatipestifer （sreotype 1,2）.

关 键 词：鸭疫里默杆菌 mazp基因 免疫原性 保护指数 

分 类 号：Q785[生物学—分子生物学] S852.617[农业科学—基础兽医学]