红豆杉细胞蛋白质双向电泳样品制备方法的比较

目的]探索适用于红豆杉愈伤蛋白质组学研究的样品制备方法。方法]以红豆杉愈伤组织为材料，分别采角三氯乙酸（TCA）／丙酮沉淀法、Tris—HCl法和酚-甲醇／醋酸铵沉淀法提取其中的总蛋白，通过双向电泳（2DE）检测3种方法对总蛋白的提取效果。结果]3种方法提取的蛋白样品2DE图谱显示的蛋白点数量均较多。TCA／丙酮沉淀法对蛋白质的提取率较高，2DE图像清晰，条纹少，拖带现象轻；Tris—HCl法提取的蛋白样品2DE图谱个别区域出现较轻的云片状聚集，并有轻微拖带现象；酚-甲醇／醋酸铵沉淀法提取蛋白样品的2DE图谱有条纹和拖带现象。结论]TCA／丙酮沉淀法提取的蛋白质质量较高，所得图谱背景清晰，蛋白质信息量大。

Comparison of Protein Sample Preparation Methods from Taxus Cell for Two-dimensional Electrophoresis

Objective] The aim was to study protein sample preparation method that suitable for proteomics research of taxus callus.Method] With taxus callus as the materials,its total protein was extracted by trichloroacetic acid(TCA)/acetone precipitation,Tris-HCl and phenol-methanol/ammonium acetate precipitation methods,the extraction effects of the 3 methods on total protein were detected by two-dimensional electrophoresis(2DE).Result] Protein spots on 2DE map of total protein extracted by 3 methods were all mo...