不结球白菜雌蕊蛋白质双向电泳技术体系的建立

以不结球白菜品种‘矮脚黄’的雌蕊为材料,从蛋白质提取、IEF程序、IPG胶条pH范围和染色方法等方面进行比较,利用Tris-HCl提取方法,使用pH 4～7 IPG胶条和改进的等电聚焦程序,改良银染方法,建立了适用于不结球白菜雌蕊蛋白质的双向电泳技术体系,并使用该体系对开花前2 d的花蕾雌蕊和开花2 d后的雌蕊蛋白质进行比较分析,获得187个差异蛋白点。

Establishment of two-dimensional gel electrophoresis system for protemics analysis of pistils in non-heading Chinese cabbage

To establish a proper two-dimensional gel electrophoresis(2-DE) system for pistils of Brassica campestris ssp.chinensis Makino,several protein extraction methods,isoelectric focusing procedures and silver staining methods were compared.Consequently,the total proteins were extracted from non-heading Chinese cabbage’Aijiaohuang’pistils by the tris(hydroxymethyl)aminomethane-hydrochloric acid(Tris-HCl)extraction method.The proteins were separated by a improved isoelectric focusing procedure,with 17 cm,pH 4-7 IPG strips and the 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE).The 2-DE map was dyed by the improved silver staining method.The different protein expression by the previous established protocol between bud pistils 2 days before florescence and blossom pistils 2 days after florescence was compared,and 187 distinct protein spots were detected.