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【目的】比较鱼腥草叶片蛋白质提取方法,建立适用于鱼腥草蛋白质组双向电泳分析方法。【方法】以鱼腥草叶片为材料,分别采用凯基试剂盒、三氯乙酸/丙酮沉淀法、尿素/硫脲提取法及酚法4种方法提取鱼腥草叶片总蛋白,使用Bradford法测定蛋白质浓度,计算蛋白提取率,并对所提取的蛋白样品进行聚丙烯酰胺凝胶电泳(SDS-PAGE)及双向电泳(2-DE)分析,所得2-DE凝胶图片采用PDQuest软件进行分析。【结果】4种方法的提取率分别为2.90、2.55、3.90、3.80 mg/g,分别检测到186、153、356、315个蛋白点,酚法提取蛋白质的2-DE图谱效果较好,结果的重复性较好。【结论】不同方法提取鱼腥草叶片总蛋白的2-DE结果存在差异,以酚法提取的2-DE效果较好,适用于鱼腥草叶片蛋白质组分析。
【Objective】 The purpose of this study was to compare the protein extraction methods of Houttuynia cordata with two-dimensional gel electrophoresis. 【Method】 Houttuynia cordata leaves were used as materials to extract the total protein of Houttuynia cordata Thunb. Leaves by Kelikin kit, trichloroacetic acid / acetone precipitation method, urea / thiourea extraction method and phenol method respectively. The total protein was determined by Bradford method Protein concentration was calculated to calculate the protein extraction rate, and the extracted protein samples were analyzed by polyacrylamide gel electrophoresis (SDS-PAGE) and two-dimensional electrophoresis (2-DE), and the obtained 2-DE gel images were analyzed by PDQuest software. 【Result】 The results showed that the extraction rates of the four methods were 2.90,2.55,3.90 and 3.80 mg / g, respectively, and 186,153,356 and 315 protein spots were detected respectively. The 2-DE pattern of protein extracted by phenol was better. The results The repeatability is better. 【Conclusion】 The 2-DE results of different methods for extracting total protein of Houttuynia cordata Thunb leaves were different. The 2-DE extracted by phenolic method was better and was suitable for the proteome analysis of Houttuynia cordata.