布鲁氏菌外膜蛋白免疫蛋白质组学方法的建立  

Application of immunoproteomics in research on outer membrane protein of Brucella

作　　者：吴朔[1] 赵忠鹏[2] 罗德炎[2] 叶艺[1] 王希良[2] 于三科[1] 

机构地区：[1]西北农林科技大学动物医学院,陕西杨陵712100 [2]军事医学科学院微生物流行病研究所免疫室病原微生物生物安全国家重点实验室,北京100071

出　　处：《免疫学杂志》2008年第4期385-388,共4页Immunological Journal

基　　金：国家自然科学基金(30170853)

摘　　要：目的建立布鲁氏菌外膜蛋白的免疫蛋白质组学研究方法,筛选布鲁氏菌保护性抗原。方法利用双向电泳技术对实验条件下培养的布鲁氏菌疫苗株M5外膜蛋白进行分离,结合WB(Western blotting)技术寻找发生免疫反应的蛋白质。结果21个免疫蛋白点经胶内酶切、肽提取后用基质辅助激光解析/电子飞行时间质谱(MALDI-TOF-MS)进行鉴定。每个蛋白点的肽质量指纹图谱用Mascot进行检索后,代表了12个开放阅读框。这些蛋白不全是外膜蛋白,还存在胞浆蛋白,其功能涉及生物合成和物质代谢等领域,还有一个功能未知的蛋白。结论成功建立了布鲁氏菌外膜蛋白的免疫蛋白质组学研究方法,为寻找保护性抗原及为新型疫苗抗原候选提供新思路。Objective To evaluate the application of immunoproteomic methods in research on Brucella vaccine strain M5. Methods Two-dimension electrophoresis was used to separate the outer membrane protein of Brucella vaccine strain M5 grown under laboratory condition, and then Western blotting was also used to semen immunogenic proteins. Results After in-gel digestion, twenty-one immunoreac- tive spots were identified by matrix-assisted laser desorption ionization-time of flight mass spectrometry （MALDI-TOF-MS）. All the peptide mass fingerprints （PMFs） were searched by the program of Mascot developed from Matrix science. These proteins represented 12 discrete open reading frames （ORFs）. These proteins were not only outer membrane proteins, but also endochylema proteins. Some of them were involved in protein synthesis and protein fate, and some of them were related substance metabolism, while function of one protein was unknown. Conclusion The immunoproteomic methods can be successfully applied in identifying immunogenic proteins of Brucella vaccine strain M5, which helps to search for other immunogenic proteins and lay a foundation for searching vaccine candidate antigens in future.

关 键 词：布鲁氏菌 外膜蛋白 双向电泳 质谱 免疫蛋白质组学 

分 类 号：R392[医药卫生—免疫学]