Abstract: [Objective] To explore the extraction methods of protein samples for SDS-PAGE and two-dimensional electrophoresis (2-DE) of the stem, vegetable, and vegetable seeds, and lay the foundation for the study of seed protein proteomics of the two species of Lepidium plants.[Method]The efficiency of four protein extraction methods were compared and analyzed through protein purity detection and SDS-PAGE.[Result]Of all the four methods, TCA/acetone extraction method generated higher yield as well as highest resolution and clear estelectrophoresis bands in SDS-PAGE, lysis extraction method got the highest protein yield whereas with highest impurities, Tris-HCl method and phenol method could obtain the highest purity but lowest protein yield, meanwhile protein extracted by lysis method and phenol method had the highest level of high-abundance proteins which could cause serious interference in further SDS-PAGE and 2-DE analysis.We got 6 differential expressed bands after compared between two species of Lepidium plants by SDS-PAGE using seed total protein extracted by TCA/acetone extraction method and phenol method.[Conclusion]TCA/acetone seed total protein extraction method is an optimal sample preparation protocol for SDS-PAGE and 2-DE.Besides, the differential bands of SDS-PAGE electrophoresis of two kinds of Lepidium seed protein could provide reference for screening two seeds.

摘要： [目的]研究适用于抱茎独行菜和独行菜种子SDS-PAGE电泳和双向电泳的蛋白样品提取方法,为研究两种独行菜种子蛋白组学奠定基础.[方法]采用4种蛋白质提取方法提取种子总蛋白,测定和分析蛋白产量和SDS-PAGE电泳图谱.[结果]TCA/丙酮提取法获得蛋白量较多,杂质少,电泳条带多且清晰;裂解液提取法蛋白的产量最高,有拖尾且杂质多;Tris-HCl提取法和苯酚提取法的蛋白产量低,杂质少;裂解液提取法和苯酚提取法获得的蛋白在SDS-PAGE电泳时易产生高丰度蛋白的干扰.对比TCA/丙酮提取法和苯酚提取法获得的两种独行菜种子总蛋白SDS-PAGE图谱,发现有6条蛋白差异条带.[结论]TCA/丙酮提取法提取两种独行菜种子总蛋白效率高,纯度高且杂质少,可用于SDS-PAGE电泳和双向电泳分析;两种独行菜种子蛋白SDS-PAGE电泳的差异谱带可为甄别两种独行菜种子提供参考依据.