几种不同提取方法对小麦叶片总蛋白双向电泳的影响

Effect of Different Extraction Protein Methods on Two-dimensional Electrophoresis of Wheat Leaf Protein

为探索小麦叶片蛋白质提取的最优方法,以普通小麦品种石4185的叶片为材料,研究了三氯乙酸/丙酮法、裂解液法、尿素/硫脲法、KCl-醋酸铵甲醇法等四种不同蛋白质提取方法对小麦叶片蛋白双向电泳图谱的影响。结果表明,三氯乙酸/丙酮法获得的蛋白点形状规则、清晰,且重复性好;裂解液法拖尾现象较轻,但蛋白点数少且欠清晰,重复性也不如前者;尿素/硫脲法的蛋白点数很少,也较为弥散,且有明显的拖尾现象;KCl-醋酸铵甲醇法使大量蛋白丢失,蛋白点数非常少,且上部竖纹干扰较重。因此,三氯乙酸/丙酮法是提取小麦叶片蛋白的较优方法。

In order to study the effect of different protein extraction methods on two-dimensional electrophoresis（2-DE） and explore an optimal protein extraction method of wheat leaves, we compared the effect on 2-DE of 4 different protein extraction methods, including TCA/acetone method, lysis-buffer method, urea/thiourea method and KCl-ammonium acetate-methanol method, using leaves of wheat cultivar Shi-4185. The results indicated that the optimal method was TCA/acetone method, which had stable, clear and repeatable protein spots. The second was lysis-buffer method with slight tailing phenomena, and lower the repeatability and clarity. The 2-DE of urea/thiourea method has fewer and dispersed protein spots. The KCl-ammonium acetate-methanol method was the worst with more impurities and less protein spots. The results showed that the TCA/acetone method is the most suitable protein extraction method for wheat leaves .