【摘要】 通过对野牛草种子蛋白双向电泳蛋白样品制备方法、等电聚焦程序以及固化胶条pH范围优化筛选,构建了适合其植物种子蛋白组研究的电泳体系。实验结果表明,使用Trizol法制备样品,可得到高纯度种子蛋白,结合使用pH4～7IPG胶条,以及优化的聚焦程序Ⅲ,能获得高分辨率、重复性好的蛋白质2-DE图谱。图像分析表明,银染后可以检测到600个以上蛋白点,其等电点主要分布于pH4～7区域,分子量主要分布于10～120kDa之间。更多还原

【Abstract】 Through optimization and screening of two-dimensional electrophoresis sample preparation methods of seeds protein of Buchloe dactyloedes(Nutt.) Engelm,the procedure of isoelectric focus and the pH range of solidifying strips,electrophoresis system that was suitable to seed protein of Buchloe dactyloedes was structured.The results showed that highly pure seed protein can be obtained with Trizol method,2-DE images of protein of high resolution ratio and good repetitiveness can be got with optimized isoelectric focus procedure.The image analysis showed that more than 600 protein spots per gel under silver staining can be detected,isoelectric points(pI) ranged mainly from pH4 to pH7 and molecular weights(MW) from 10 kDa to 120 kDa.更多还原