【摘要】 为了筛选和鉴定猪链球菌3个保护性抗原GAPDH、MRP、DLDH的优势B细胞抗原表位,设计表位串联方案,研究重组串联表位蛋白(简称GMD蛋白)免疫小鼠后对猪链球菌攻击的免疫保护效力。采用生物信息学分析工具对优势B细胞抗原表位进行筛选,并将表位串联并构建重组质粒pET28a-GMD,诱导表达且纯化后将GMD蛋白配合弗氏佐剂免疫BALB/c小鼠,并评价该蛋白对2型猪链球菌的免疫保护效力。发现经SDS-PAGE鉴定该重组蛋白大小为43.3 kDa;小鼠免疫保护实验结果显示,免疫小鼠血清中针对猪链球菌HA9801菌株抗体效价最高可达1∶102 400,GMD蛋白对小鼠的免疫保护率可达90%。上述结果表明面对猪链球菌HA9801菌株的攻击,GMD蛋白可以为机体提供良好的免疫保护,本研究结果为猪链球菌新型表位亚单位疫苗的研制提供了基础数据。更多还原

【Abstract】 Streptococcus suis is an important pathogen causing pig diseases.However,there is no effective vaccine providing cross protection against multi-serotype S.suis.In this study we constructed a subunit vaccine that targeting the conservative protective antigens of S.suis. The B cell dominant epitopes of three important protective antigens(GAPDH,MRP,DLDH) of S.suis were analyzed and a strategy to express the tandem protein named GMD was designed containing 3 dominant B cell epitopes of GAPDH,8 dominant B cell epitopes of MRP,and 4 dominant B cell epitopes of DLDH.The immunogenicity of GMD was evaluated in mice.After three doses immunization,the antibody titer against S.suis strain HA9801 was up to 1∶102 400.Seven days after the last immunization,the mice were challenged with HA9801.The survival rate of GMD immunized mice reached 90%.Statistically protection against S.suis was found to have been achieved.更多还原