SS2溶菌酶释放蛋白(MRP)B细胞表位预测、原核表达及免疫学特性  

The prokaryotic expression and immune characteristic analysis of predicted B cell epitope in streptococcus suis type 2(SS2) lysozyme release protein(MRP)

作　　者：吴立志[1,2] 伍晓雄[2] 张松林[1] 刘磊[1] 肖跃强[1] 夏小静[1] 沈志强[1] 

机构地区：[1]山东省滨州畜牧兽医研究院,山东滨州256600 [2]华中农业大学动物医学院,湖北武汉430070

出　　处：《中国兽医学报》2013年第4期571-575,580,共6页Chinese Journal of Veterinary Science

基　　金：山东省滨州畜牧兽医研究院自主创新基金资助项目(201002);山东绿都生物科技有限公司技术创新资助项目(201003)

摘　　要：运用ANTHEPROT 5.0综合分析二级结构、亲水性、表面可及性与抗原性指数,预测MRP的B细胞优势表位簇;PCR扩增出表达预测的抗原肽段的基因(mrp-1),构建重组原核表达载体pET32a-mrp-1,IPTG诱导表达,以渗透性休克法纯化重组蛋白;使用重组蛋白免疫血清和灭活猪链球菌2型SS2免疫血清Western blotting检测重组蛋白的免疫原性及其与SS2免疫血清的反应性;纯化的重组蛋白免疫雌性SPF昆明小鼠,SS2攻毒受试动物评估重组蛋白对小鼠的免疫保护力。Western blotting显示预测的950～1 210aa肽段与PET32a重组表达的蛋白(MRP)具有良好的免疫原性和反应性;攻毒试验结果表明重组蛋白对小鼠的免疫保护率为菌体免疫所提供保护率的66.7%。研究为利用该细胞表位簇作为高效免疫制剂奠定了基础。Get the prepotent antigenic determinant by using ANTHEPROT 5.0 to analyse the secondary structure, hydrophily, accessibility and antigenicity of MRP; The predicted 950-1 210 aa peptide（MRP-1） prokaryotic expression vector pET32a-mrp-1 was constructed by using genetic engineering method and then purified by osmotic shock ; analyse the immunogenicity and reactivity by Western blotting analysis using the purified MRP-1 incubation with MRP-1 immune serum and SS2 immune serum, respectively. Observe the role of phylaxis in SS2 infection through the mice survival rate after using the purified protein MRP-1 immune mice and injecting the fatal dose of SS2. Western blot displayes that the recombinant protein has good immunogenicity and reactivity; animal experiment results shows that the recombinant protein can protect 60% mice against the infection of SS2. This study laid foundation for the applicaton of this protein as a novel efficient immune reagent.

关 键 词：猪链球菌2型 溶菌酶释放蛋白 B细胞表位 原核表达 免疫保护 

分 类 号：S852.611[农业科学—基础兽医学]