番茄果实中蛋白质的提取和双向电泳条件的优化

Improvement of extraction method and two-dimensional electrophoresis conditions for proteome study of tomato fruit (Lycopersicon esculentum)

由于果实组织中富含多糖、多酚和色素等物质,导致了果实组织比其他植物组织蛋白质的提取要更困难。研究以番茄果实为模式材料,对番茄果实蛋白样品制备、蛋白裂解缓冲液、上样量等方面作了改进,应用ImageMaster2DElite软件分析对银染的图谱进行分析,凝胶上检测到超过800个的蛋白质,不同凝胶之间蛋白质的匹配率达到95%,具有较高的分辨率和重复性,建立了一套适用于番茄果实蛋白质组分析的双向电泳(2-DE)方法。

The isolation of protein from tomato fruit tissues rich in polysasccharide, polyphenol and pigments has been a long-standing problem for researchers. In this study, protein sample preparation, lysis buffer and loading of samples were modified and improved to minimize the interference substance in tomato fruit. More than 800 proteins were detected in each gel and the 95% of them were matched with each other in different silver stain gels analyzed by Image Master 2D Elite software, indicating the protocol with high repeatability and resolution. 2-DE （two-dimensional electrophoresis） protocol suited for the separation of proteins from tomato fruit was established.