外膜蛋白OmpC在枯草芽孢杆菌中的分泌表达及重组活菌制剂在肉鸡上的应用

发布时间：2018-04-26 00:09

本文选题：沙门菌 + 外膜蛋白　；参考：《畜牧与兽医》2017年08期

【摘要】：为了研究携带鸡白痢沙门菌主要外膜蛋白OmpC基因的重组枯草芽孢杆菌活菌制剂在肉鸡上的免疫原性,以具有益生菌属性的枯草芽孢杆菌作为基因疫苗表达宿主,构建表达鸡白痢沙门菌主要外膜蛋白的枯草芽孢杆菌重组菌(pHT43/WB800n-OmpC)。将重组菌饮水饲喂12日龄艾维茵肉鸡,同时设空白、空转菌及非诱导菌对照组,分别于免疫后27和42日龄测定特异性抗体效价,同时45日龄后采用沙门菌C79-13株腹腔注射攻毒。结果:SDS-PAGE分析显示该重组蛋白大小为37 ku;Western blot分析显示表达蛋白具有很好的反应原性;肉鸡饲喂结果显示,42日龄时血清中检测到特异性抗体,且抗体效价为1∶32;攻毒试验显示重组菌诱导组免疫保护率为76.92%,而对照组、空转菌组、非诱导组保护率分别为23.08%、38.46%和30.77%。结果表明:此重组菌对鸡沙门菌具有免疫保护作用,为沙门菌的免疫机制研究奠定了基础,同时实现了饲用疫苗的双重作用。
[Abstract]:In order to study the immunogenicity of recombinant Bacillus subtilis OmpC gene carrying the main outer membrane protein of Salmonella pullorum on broiler chickens, Bacillus subtilis with probiotic properties was used as the expression host of the gene vaccine. The recombinant Bacillus subtilis strain expressing the main outer membrane protein of Salmonella pullorum (Salmonella pullorum) was constructed. The recombinant bacteria were fed to 12 day old Avian broilers in drinking water. The control group was divided into two groups. The specific antibody titers were determined at 27 days and 42 days after immunization respectively. After 45 days of age, Salmonella C79-13 strains were injected intraperitoneally to attack the virus. Results the specific antibody was detected in the serum of broiler chickens at 42 days of age by the analysis of the recombinant protein with the size of 37 Ku blot, and the results of broiler feeding showed that the expressed protein had a good reactiveness, and the specific antibody was detected in the serum of broiler chickens at the age of 42 days. The antibody titer was 1: 32, and the protective rate of recombinant bacteria was 76.92, while that of control group, idle group and non-induction group was 23.088.46% and 30.77%, respectively. The results showed that the recombinant strain had immune protection against Salmonella chickens. it laid a foundation for the study of immune mechanism of Salmonella and realized the double function of feed vaccine.
【作者单位】：成都农业科技职业学院畜牧兽医分院;四川农业大学动物医学院;
【分类号】：S858.31

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