苔藓植物双向凝胶电泳蛋白样品制备的优化

比较了苔藓植物小立碗藓3种蛋白样品制备方法,结果分析表明,直接研磨法和三氯乙酸（TCA）/丙酮（acetone）沉淀法检测的蛋白点的数量相差不明显;直接研磨法提取蛋白所得到的2-DE图谱蛋白点模糊,分辨率低,横竖条纹干扰严重;三氯乙酸（TCA）/丙酮（acetone）沉淀法分离效果较好,分辨率有所提高.改良的酚抽提/乙酸铵沉淀法检测的蛋白点远远多于前两种方法,所得到的图谱质量最高,分辨率大大提升,蛋白点分布均匀清晰,横竖条纹干扰也很少,凝胶背景干净,是最佳的小立碗藓蛋白样品制备的方法.

Optimization of sample preparation for two-dimensional gel electrophoresis in bryophytes

Two-dimensional gel electrophoresis is a powerful method for protein separation in proteomics research.The preparation of protein samples is critical to acquire a good two-dimensional electrophoresis profile.It needs different sample preparation techniques for different types and sources of protein samples.To find the most efficient method for sample preparation,the different methods were analyzed in the moss Physcomitrella patens.There is no significant difference in the protein spots between direct grinding method and trichloroacetic acid/acetone precipitation method.The 2-DE profile of trichloroacetic acid/acetone precipitation method is better than that of direct grinding method.Phenol extraction/ammonium acetate precipitation is the best sample preparation method for Physcomitrella patens.