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棉花学报  2008 

蛋白质组学研究中的质谱鉴定与生物信息学分析

DOI: 1002-7807(2008)04-0281-08, PP. 281-288

Keywords: 棉花,胚珠,纤维,蛋白质,质谱

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Abstract:

以陆地棉TM-1开花后6d的胚珠为对照,比较开花后6、10、14、18、22d纤维的蛋白质组双向电泳图谱,从中选取了编号为37的差异表达的蛋白质点。胶内胰蛋白酶酶切的多肽,用AB4700蛋白质组学分析仪进行MALDI-TOF/TOF分析,获得了该点内蛋白质的高品质肽质量指纹图谱(PMF),从中挑出质量为1517.9的肽离子进行串联质谱分析,获得碰撞后断裂片段离子的串联质谱(FFP)。将获得的PMF分别用MASCOT、ProFound、Aldente和MS-Fit等常用软件对NCBInr和UniProt数据库中的绿色植物蛋白质数据库进行搜索,并用MASCOT搜索本地棉属EST数据库。将获得的FFP用MASCOT软件的离子搜索模式和序列查询模式搜索数据库结合手工质谱解析推断序列,最后确认该蛋白质是羟甲基转移酶。本文着重讨论了蛋白质组学研究中质谱分析和生物信息学软件的应用,评价常用蛋白质组学PMF检索工具及其检索结果。

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