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水稻是研究单子叶植物遗传和分子生物学的模式植物,是研究植物基因组学和蛋白质组学的一个重要的生物材料。双向电泳作为蛋白质组学中蛋白分离的主要方法,其关键步骤在于蛋白质的提取。为了比较不同蛋白提取方法,采用TCA/丙酮沉淀法和Mg/NP-40提取法分别提取水稻叶片总蛋白,用双向凝胶电泳分离两种方法获得的蛋白。结果表明,在双向图谱中,Mg/NP-40提取法分离的蛋白点数较多,RuBisCO大亚基的含量较低,适于分离等电点在5～7范围内、分子量在14～20kD和高于67kD的水稻叶片蛋白质;TCA/丙酮沉淀法分离的蛋白点数较少,适于分离等电点在4～5范围内、分子量在31～67kD内的水稻叶片蛋白质。
Rice is a model plant for studying the genetics and molecular biology of monocotyledons and is an important biological material for studying plant genomics and proteomics. Two-dimensional electrophoresis as the main method of protein separation in proteomics, the key step is the protein extraction. In order to compare different protein extraction methods, TCA / acetone precipitation method and Mg / NP-40 extraction method were respectively used to extract the total protein of rice leaves and the two proteins were separated by two-dimensional gel electrophoresis. The results showed that Mg / NP-40 extraction method could separate more protein spots and RuBisCO large subunit in the two-way map, which was suitable for separating isoelectric point in the range of 5 ~ 7 and molecular weight in 14 ~ 20kD and Higher than 67kD of rice leaf protein; TCA / acetone precipitation protein spots isolated less suitable for separating isoelectric point in the range of 4 to 5, the molecular weight of 31 ~ 67kD within the rice leaf protein.