棉花根总蛋白双向电泳方法的优化  

Optimization of Two-Dimensional Electrophoresis Methods for Separating Cotton Root Total Proteins

作　　者：孙立荣[1] 胡盼盼[1] 李亚平[1] 王晓阳[1] 郝福顺[1] 

机构地区：[1]河南大学棉花生物学国家重点实验室植物逆境生物学重点实验室生命科学学院,河南开封475004

出　　处：《河南大学学报（自然科学版）》2014年第3期329-333,共5页Journal of Henan University:Natural Science

基　　金：棉花生物学国家重点实验室专项经费资助

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.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.

关 键 词：陆地棉 蛋白质提取 双向电泳 蛋白质组学 

分 类 号：Q816[生物学—生物工程]