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  

Evaluation of application of recombinant Bacillus Subtilis expressing Salmonella pullorum OmpC gene in broiler chickens

作　　者：王振华[1] 潘康成[2] 包小玉 

机构地区：[1]成都农业科技职业学院畜牧兽医分院,四川温江611130 [2]四川农业大学动物医学院,四川温江611130

出　　处：《畜牧与兽医》2017年第8期84-87,共4页Animal Husbandry & Veterinary Medicine

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。To investigate the immune efficacy of recombinant Bacillus subtilis carrying the outer membrane protein OmpC gene of Salmonella pullorum,B. subtilis WB800 n was selected as host bacteria. Then the 12-day-old broilers chickens were orally inoculated with the recombinant B. subtilis（ pHT43/WB800n-OmpC）,and B. subtilis WB800 n and no-inducible B. subtilis recombinant bacteria served as controls.The titer of specific antibody against S. pullorum was determined at 27 and 42 days after immunization,and 45 days later,S. pullorum C79-13 was challenged intraperitoneally. The result showed that the recombinant B. subtilis pHT43/WB800n-OmpC expressing OmpC gene was successfully constructed. The molecular weight of the expressed recombinant protein was determined to be about 37 ku by SDS-PAGE. Western blot confirmed that the recombinant protein had good reactogenicity. The serum specific antibodies were detected at 42 days after immunization and the average antibody titer was 1 ∶ 32. The protection rate against S. pullorum was 76. 92% for the experimental group immunized with the recombinant B. subtilis pHT43/WB800n-OmpC,while 23. 08%,38. 46% and 30. 77% for the blank control group,B. subtilis WB800n group and the no-inducible B. subtilis recombinant bacteria,respectively. It is concluded that the OmpC gene of Salmonella was firstly expressed in B. subtilis,which lay a theoretical foundation for investigation of Salmonella immune mechanism and for development of genetic engineering probiotics that play a dual role after oral immunization.

关 键 词：沙门菌 外膜蛋白 OMPC 枯草芽孢杆菌 免疫保护率 

分 类 号：S852.6[农业科学—基础兽医学]