2型猪链球菌菌毛结构亚蛋白tSSU0473的原核表达与免疫学特性研究  

Prokaryotic Expression and Immunological Characterization of Pilus Subunit tSSU0473 of Streptococcus suis Serotype 2

作　　者：胡丹[1] 张先云[1] 耿美玲[1] 张凤玉[1] 郝丽娜[1] 朱静[1] 郑峰[1] 潘秀珍[1] 王长军[1] 

机构地区：[1]南京军区军事医学研究所,江苏南京210002

出　　处：《生物技术通讯》2013年第6期753-758,共6页Letters in Biotechnology

基　　金：国家自然科学基金(31170124;30972638;81071317;81171527;81172794);江苏省自然科学基金(BK2011097;BK2010025;BK2010114;BK2010113)

摘　　要：目的:探究2型猪链球菌(S.suis2)强毒株05ZYH33的srtF基因簇编码的菌毛结构亚蛋白SSU0473的细菌定位及其免疫保护效能。方法:原核表达截短的SSU0473(tSSU0473),并以亲和层析法纯化目的蛋白,Western印迹检测tSSU0473蛋白的免疫原性,ELISA法检测多抗血清的效价及IgG亚型,小鼠试验测试重组蛋白的免疫保护效能,免疫电镜观测tSSU0473蛋白的细菌定位。结果:在原核系统中表达了tSSU0473蛋白;ELISA结果显示重组蛋白能够刺激小鼠产生高效价的免疫抗体;动物试验表明tSSU0473蛋白免疫小鼠可抵御致死剂量病原体的攻击,显示出较好的免疫保护作用;免疫电镜检测显示tSSU0473蛋白定位于细菌表面。结论:菌毛亚蛋白tSSU0473是S.suis2膜表面蛋白,具有良好的免疫原性和免疫保护性,可作为S.suis2亚单位疫苗的候选分子。研究结果为系统揭示S.suis2的菌毛生物学结构与功能奠定了基础。Objective： To elucidate the inmmunoprotection of pilus subunit SSU0473 encoded by the srtF pilus island in the Chinese strain 05ZYH33 of Streptococcus suis serotype 2（S.suis2）. Methods： The truncated protein tSSU0473 was expressed in prokaryotic cells and was purified by affinity chromatography. Then recombinant protein was analyzed by Western blotting, and the anti-SSU0473 serum was prepared by immunizing mice with recombinant tSSU0473 protein and analyzed by ELISA. The location site of tSSU0447 protein in bacterium was detected by immune electron microscopy. Results： The tSSU0473 protein was expressed successfully in prokaryotic system. The high titer of the serum was obtained, ELISA result indicated that the recombinant protein can react with mice anti-SSU0473 serum. Animal test showed that vaccinating mice with recombinant tSSU0473 protein conferred a significant protection. The tSSU0473 protein was detected on bacterial surface by immune electron microscopy. Conclusion： Protein tSSU0473 could serve as a vaccine candidate of S.suis2, and provides the foundation for systematically illustrating the role that pilus island plays in pathogenicity of S.suis2.

关 键 词：2型猪链球菌 菌毛 SSU0473蛋白 原核表达 免疫保护 

分 类 号：Q78[生物学—分子生物学] R392.1[医药卫生—免疫学]