双向电泳比较两种方法获得的水稻叶片蛋白

Proteins from rice leaves extracted with Mg/NP-40 buffer are different from those with TCA buffer

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 （Oryza sativa） , an established model system for monocots, is used widely in studying genomics and proteomics. Two-dimensional eleetrophoresis （2-DE） , the most effective separation strategy for protein mixture, depends heavily on protein extrac- tion. In order to compare different proteins extracted methods, both proteins of rice leaves extracted with TCA buffer and Mg/NP-40 buffer were detected with 2-DE analysis. The results indicated that more proteins could be extracted and less RubisCO tested with Mg/ NP-40 buffer than those with TCA buffer. Additionally, it was found that Mg/NP-40 buffer is better for separating 14-20kD or 〉 67kD proteins with pI 5-7, while TCA buffer seems to be better for separating 31-67kD proteins with pi4-5.