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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800μg,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。
In this study, hybrids of Sorghum sudanense and its parents were used as materials to extract the total protein of Sorghum sudanense leaves by two methods (acetone precipitation with trichloroacetic acid and precipitation with acetone). The pH range, The sample size has been optimized. The results showed that acetone precipitation method was more suitable for extracting total protein from leaves of Sorghum bicolor. Compared with acetone precipitation method of trichloroacetic acid, acetone precipitation method was used to extract total protein from leaves of Sorghum bicolor. Using 17 cm pH 4-7 IPG strips, High rate and good reproducibility of the Sorghum bicolor two-dimensional electrophoresis map, for the latter Sorghum differential proteomics study provides a good technical support.