孙强正,罗霞,叶长芸,肖迪,郑翰,景怀琦,徐建国.猪链球菌分泌蛋白的免疫蛋白质组学研究[J].中华流行病学杂志,2008,29(3):267-271 |
猪链球菌分泌蛋白的免疫蛋白质组学研究 |
Immunoprotemic assay of secretive proteins from Streptococcus suis type2 strain SC84 |
收稿日期:2007-12-14 出版日期:2014-09-15 |
DOI: |
中文关键词: 猪链球菌 分泌蛋白 免疫蛋白质组学 |
英文关键词: Streptococcus suis Secreted protein Immunoprotemics |
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中文摘要: |
目的 寻找猪链球菌(S.suis)SC84菌株分泌蛋白中具有抗原活性的蛋白,为S.suis特异诊断抗原和新的疫苗候选抗原的筛选提供线索.方法 采用免疫蛋白质组学和介质辅助激光解吸电离飞行时问质谱(MALDI-TOF MS)技术,分析和鉴定S.suisSC84菌株分泌蛋白中能与感染病例恢复期血清发生免疫杂交的蛋白点.结果 在S.suis SC84菌株分泌蛋白的免疫杂交膜上,共发现14个具有抗原活性的蛋白点,其中11个点可以在考马斯蓝染色胶上找到匹配点.11个蛋白点经质谱鉴定,分属于8种蛋白,全部定位于胞壁或胞外.其中溶菌酶释放蛋白、溶血素、细胞外因子为S.suis 已知的抗原蛋白;5'-核苷酸酶、ribonucleases G和E、金属内肽酶等为新发现的具有抗原活性的蛋白;所有的蛋白均能在测序菌株S.suis 05ZYH33基因组中找到相应的编码基因.结论 在S.suis 分泌蛋白中发现了新的具有抗原活性的蛋白,可能作为S.suis特异诊断抗原和疫苗候选抗原,用于诊断试剂和疫苗的研究 |
英文摘要: |
Objective To identify antigenic proteins secreted 6y Streptococcus suis(S.suis)type2 strain SC84. Methods Two-dimensional electrophoresis ( 2-DE ),western-blot assay and matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS) analysis were performed to search and identify antigenic proteins secreted by S.suis strain SC84, which triggered an outbreak of the disease in Sichuan province,China, in 2005. Results A total number of 14 western blot spots were found on PVDF membrane. 11 spots which could be found the existence of matching protein on coomassie G-250- stained 2-DE gel were identified by MALDI-TOF MS. The 11 proteins, all located at extra-cellular or cell wall, were classified into 8 kinds of proteins. Among of them, muramidase-released protein(MRP ), suilysin (Sly) and extra-cellular factor (EF) were the known antigenic proteins, but several proteins such as putative 5}-nucleotidase, ribo-nucleases G and E, and predicted metal-loendo-peptidase were newly found
antigenic proteins. All the identified protein were found to have had the coding gene in genomic of S.suns strain OSZYH33,isolated from patients in Sichuan province, China in 2005. Conclusion The newly found proteins could be used as voluntary antigens for detection and vaccination of S.suis. |
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