【摘要】 以陆地棉(Gossypium hirsutumL.)根为材料,比较分析了三种总蛋白提取方法及电泳参数和上样量等因素对双向电泳结果的影响.结果表明:选择pH 4~7IPG胶条,采用丙酮沉淀/2-D Clean-up Kit法提取总蛋白,电泳时设置合适的电压,上样量为250μg蛋白进行双向电泳能够检测到1 000多个清晰的蛋白点,适合做质谱分析.更多还原

【Abstract】 Effects of three isolation and purification methods of cotton(Gossypium hirsutumL.)root proteins on the two-dimensional polyacrylamide gel electrophoresis(2D-PAGE)were investigated.Some factors including electrophoretic parameters and loading amount of protein samples for the 2D-PAGE were also optimized.The results showed that acetone precipitation/2-D Clean-up kit were the best method for obtaining the total proteins from cotton roots.We obtained more than 1 000clear cotton protein pots in a 2D-PAGE map when used a pH 4-7IPG strip, suitable voltage,and uploaded 250μg protein each well.These proteins well suited for mass spectrometric analysis. These results suggest that the optimized method is suitable for studying cotton proteomics.更多还原