玉米叶片2种总蛋白质提取方法的比较

摘要

为比较不同方法提取的玉米叶片蛋白质对双向电泳效果的影响。采用TCA-丙酮沉淀和TRIzol试剂盒2种方法，提取国审玉米品种郑单958叶片全基因组蛋白质，并对提取的蛋白质样品进行双向电泳。结果表明：TCA-丙酮方法提取的蛋白质一向水平聚焦电压上升缓慢且达不到程序设置的电压，而TRIzol试剂盒方法实时监测图与程序设置图相吻合；对电泳胶图分析后可知，TRIzol试剂盒法分离出的蛋白点多于900个，低丰度蛋白质得到充分显现，大多数蛋白点分子量为20～80 kDa，pH值为4～7，蛋白点的形状较规则；TCA-丙酮沉淀法分离出的蛋白点仅有350个，表明样品中蛋白质分离种类较少，高丰度蛋白质过度显现，低丰度蛋白质检出率大大降低。%To compare the effect of corn leaf protein extraction by different methods with Two-dimensional elec-trophoresis.Two methods of TCA-acetone precipitation and TRIzol Kit were used to extract the leaf genome-wide protein from maize variety Zhengdan 958 ,and the effect of protein extractions were evaluated by two-dimensional e-lectrophoresis .The results showed that the focus voltage of proteins extracted by TCA-acetone method rose slowly and could not be up to the program settings , while the Real-time monitoring map of TRIzol Kit method coincided well with the program settings .After analysis of gel by ImageMaster 2D-Platinum,more than 900 protein spots were separated by Trizol Kit method ,and the low abundance proteins displayed fully .For majority of the protein spots ,the molecular weight (MW) was 20-80 kDa,the pH value was 4-7,and the shape was regular .The protein spots sepa-rated by TCA-acetone precipitation method were 350 only,showing the less protein types separated from a sample , while the high abundance protein over expressed and the detection rate of low abundance proteins reduced greatly .