【摘要】 对赤子爱胜蚓(Eisenia fetida)表皮组织的两种蛋白质提取方法进行了比较,并对双向电泳IPG胶条pH范围,上样量条件进行了优化,结果表明:裂解液提取法提取的蛋白质量浓度较低,蛋白种类缺失,采用TCA-丙酮沉淀法提取法,可显著提高提取液蛋白质提取效果,样品中蛋白质质量浓度可达12.49μg/μL.蚯蚓表皮组织蛋白质主要分布在pH范围4～7之间,适于选用24cmpH 4～7线性梯度IPG胶条,采用考马斯亮蓝染色法时最适上样量为600μg,所得电泳图谱可检测蛋白点达(629±20)个,且重复性实验匹配率较高.用TCA-丙酮沉淀法提取蚯蚓表皮组织蛋白质组,按优化的参数进行双向电泳分离,所得图谱分辨率高、重复性好,可用于蚯蚓蛋白质组学研究.更多还原

【Abstract】 In order to establish and optimize two-dimensional electrophoresis(2-DE) technology system for earthworm Eisenia fetida epithelium proteome,sample preparation method,loading quantity of samples,appropriate IPG strips and system repetition were studied.The results showed that,compared with the lysis buffer extraction method,TCA-A extraction method could increase the protein extracting efficiency up to 12.49 μg/μL.While the 24 cm IPG strip(pH 4~7) and 600 μg loading sample quantity were considered to be the optimal condition for 2-DE analysis of earthworm Eisenia fetida epithelium proteome,up to(629±20) protein spots can be detected on the gel after coomassie brilliant blue R-250 staining,repeatability of the system also turned out to be good.Therefore,the high resolution and repeatable 2-DE technology system could be wildely used in proteome study of earthworm Eisenia fetida.更多还原