本研究以杂种高丹草及其亲本为试材,分别采用2种方法(三氯乙酸丙酮沉淀法,丙酮沉淀法)提取高丹草叶片总蛋白,并对IPG胶条的pH范围、蛋白上样量进行了优化.研究结果表明:与三氯乙酸丙酮沉淀法相比,丙酮沉淀法更适合高丹草叶片总蛋白的提取;采用17 cm pH 4～7 IPG胶条,上样量为800 μg,构建了背景清晰,分辨率高,重复性好的高丹草双向电泳图谱,为后期高丹草差异蛋白质组学研究提供良好的技术支撑.
Establishment of Two-dimensional Electrophoresis System for Sorghum-Sudangrass Hybrids Leaves
In this study,sorghum-sudangrass hybrids and their parents as experimental materials,the total proteins were extracted from sorghum-sudangrass hybrids leaves in two methods (TCA-acetone precipitation method and acetone precipitation method).The applicable strip of different pH range and loading quantities of protein were chosen.The results showed that acetone precipitation method was more suitable for protein extraction compared with TCA-acetone precipitation method.We builted a clear background,high resolution and good repeatability two dimensional electrophoresis maps of sorghum-sudangrass hybrids which using 17 cm pH 4～7 IPG strip,and loading quantities with 800 μg,which provides a good technical support comparative proteomics research of heterosis of sorghum-sudangrass hybrids.
NETL
NSTL
万方数据
维普
