大麦和麦芽水溶性蛋白提取方案的优化及双向电泳图谱的建立

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。

Optimization of extraction method of the water-soluble proteins of barley and malt and construction of two-dimensional electrophoretic map

To develop an efficient and high resolution 2-DE procedure for barley and malt research,in this study,the method of extraction and sample preparation for two dimensional gel electrophoresis(2-DE) were optimized using high quality barley cultivars Gairdner and by changing the type and amount of extraction reagents as well as extraction time.By using the optimized method,protein extraction and sample preparation methods could be achieved.The protein sample of barley was obtained after grinding,buffer-phenol extraction,ethanol washing,and was freeze-dried.Protein spots were separated by 2-DE technique,stained with silver nitrate,recorded and detected by PDQuest software.The results showed that over 700 spots of barley and about 500 spots of malt were separated by 2-DE.The established of those methods may provide powerful tools for developmental proteomics and quality improvement ofbarley in the future.