马流产沙门菌鞭毛重组蛋白FliC的原核表达及对小鼠的免疫保护试验  

Prokaryotic expression of FliC protein of Salmonella abortus equi and immune protection test in mice

作　　者：杨康 易鸳鸯[1] 赵亚南 常建新 苏艳 

机构地区：[1]新疆哈密市动物疫病预防控制中心,新疆哈密839000 [2]新疆农业大学动物医学学院,新疆乌鲁木齐830052

出　　处：《中国兽医科学》2017年第12期1549-1554,共6页Chinese Veterinary Science

基　　金：新疆维吾尔自治区重大科技专项项目(2017A01002-2);国家自然科学基金项目(31660707)

摘　　要：为研究马流产沙门菌新疆分离株鞭毛蛋白FliC(flagellin C)的免疫生物学特性及保护特性。本研究利用PCR技术扩增获得该菌的Fli C基因,并将其连接到原核表达载体p ET-28a(+)上,构建重组表达质粒,转化大肠杆菌BL21(DE3)感受态细胞后,用IPTG诱导表达重组蛋白,SDS-PAGE和Western-blot检测与分析表达蛋白,并将纯化的重组蛋白免疫小鼠。结果显示,成功诱导表达了52 ku的重组蛋白,该蛋白第2次免疫小鼠第14天时血清中的特异性Ig G抗体效价可达1∶72 900,且抗体亚型以Ig G1为主,免疫小鼠的攻击保护率可达73%。结果表明,Fli C重组蛋白具有良好的免疫原性和免疫保护效果。为进一步开展马流产沙门菌亚单位疫苗的研究奠定了基础。To investigate the immune biological characteristics and protective efficacy of Salmonella abortus equi FliC （Flagellin protein C ）, the FIiC gene was amplified by PCR method and cloned into pET-28a（ ＋ ） prokaryotic expression vector in this study.The recombinant plasmid was then transformed into Escherichia coli BL21 （DE3）.The recombinant FliC protein was expressed after being induced with IPTG and analyzed by SDS-PAGE and Western-blot.The purified FIiC protein was used to immunize mice.The results indicated that the 52 ku recombinant protein was successfully expressed and the mice IgG specific antibody titer was 1 ： 72 900 at 14 d after the second immunization.The antibody subtype of immunized mice was mainly IgGl and mice had 73% survival rate after challenge.This result indicated that the FIiC protein had good immunogenicity and induced better immune protective efficacy,which laid foundation for further studies of S. abortus equi subunit vaccines.

关 键 词：马流产沙门菌 FliC蛋白 表达与纯化 免疫保护效果 

分 类 号：S852.612[农业科学—基础兽医学]