甲型副伤寒沙门菌外膜蛋白质组学参考图谱的建立

摘要

目的 建立甲型副伤寒沙门菌外膜蛋白的蛋白质组学参考图谱.方法 甲型副伤寒沙门菌CMCC 50973培养至对数生长末期,收集菌体并破碎提取细菌的外膜蛋白,然后利用双向电泳(2-DE)将蛋白分离.分离后取蛋白点做MALDI-TOF-MS鉴定,根据质谱得到的肽指纹图谱在Mascot数据库中查询出匹配的蛋白.结果 在2-DE考马斯亮蓝染色凝胶上取77个蛋白点,鉴定成功了54个点,对应29种蛋白.29种蛋白中定位于外膜为13种,定位未知的有11种,胞质中有5种.结论 成功建立了甲型副伤寒沙门菌外膜蛋白的蛋白质组学参考图谱,为进一步筛选疫苗抗原打下了基础.%Objective To establish a proteomic reference map of outer membrane protein ( OMPs) of Salmonella paratyphi A (SPA) CMCC 50973.Methods To identify the major OMPs of SPA, proteomic analysis was undertaken using a carbonate OMP-enrichment protocol.The enriched OMP-extracts were separated by 2-dimensional eleetrophoresis and the spots were identified by MALDI-TOF-MS combined with Mascot database searches.Results A total of 54 spots in gel were identified by MALDI-TOF-MS and database searches, representing 29 protein entries.The subcellular locations of the identified 29 proteins were predicted using the software PSORT.Of the 29 proteins, 13 were predicted to be outer membrane, 11 unknown and five were cytoplasmic.Conclusion A proteomic reference map of SPA OMPs is successfully established.