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为了建立一套适用于大麦和麦芽的双向电泳分析方法,得到更加清晰、全面的双向电泳图谱。以高产优质大麦品种Gairdner为材料,对提取方法以及提取过程中所用的试剂的种类、用量和处理时间等进行比较分析和优化,运用蛋白定量法测定提取样品的蛋白质含量,摸索出一套完整的用于双向电泳分析的大麦蛋白提取和样品制备方法。大麦或麦芽经研磨,缓冲液-苯酚提取,醇洗,冻干后得到的蛋白质样品,双向电泳分离,硝酸银染色,PDQuest分析,图谱结果显示大麦检测到700个蛋白质点,麦芽检测到500个蛋白质点。这一方法的构建可作为大麦蛋白质组研究的重要工具,为今后大麦蛋白质组学以及对大麦品质改良的研究奠定了一定的基础。
In order to establish a set of two-dimensional electrophoresis analysis methods suitable for barley and malt, a more clear and comprehensive two-dimensional electrophoresis pattern is obtained. Gairdner, a high-yield and high-quality barley variety, was used to compare and optimize the extraction method, the type, amount and processing time of the reagents used in the extraction process. The protein content of the extracted samples was determined by protein quantification method, Barley protein extraction and sample preparation methods for two-dimensional electrophoresis analysis. Barley or malt milled, buffer-phenol extraction, alcohol washing, protein samples obtained after lyophilization, two-dimensional electrophoresis separation, silver nitrate staining, PDQuest analysis, the map results showed that barley detected 700 protein spots, malt detected 500 Protein spots. The construction of this method can be used as an important tool for the research of barley proteome, which lays a solid foundation for the future research of barley proteomics and the improvement of barley quality.