中文摘要：

为建立一套完整、可行的适合大豆茎秆蛋白的双向电泳技术,对大豆茎秆蛋白的提取方法、双向电泳的固相化p H梯度凝胶（IPG）、上样量、SDS-PAGE胶浓度、染色方法等进行了优化。结果表明,将传统的TCA/丙酮法与酚法相结合,提取的大豆茎秆蛋白经双向电泳分离后,可获得较多的蛋白点,且背景清晰,便以识别。双向电泳优化后的主要技术参数为p H3-10非线性IPG胶条、上样量800μg、12%SDS-PAGE胶、考马斯亮蓝染色。本研究形成的大豆茎秆蛋白提取方法和双向电泳技术方案,有利于进一步采用蛋白质组学分析大豆茎秆的生物学功能。

英文摘要：

To optimize soybean stem protein extraction and separation method for better application of 2- D electrophoresis on soybean proteomics,extraction method,p H of IPG strips,sample loading quantity,SDS- PAGE gel concentration and staining method were compared. Results showed that the new protein extraction method combining TCA / acetone precipitation and phenol extraction could resolve more protein with better separation by 2- D electrophoresis. The optimized processes included p H3- 10 non- linear IPG strip and loading 800μg soybean stem protein for the first- dimensional IEF,12% polyacrylamide gel for the SDS- PAGE electrophoresis,and Coomassie brilliant blue staining.