油茶种仁蛋白双向电泳体系的建立

Two-dimensional gel electrophoresis system establishment of Camellia oleifera kernel

以普通油茶优良无性系‘闽43’成熟种仁为材料,从样品制备、IEF聚焦程序参数设定、SDS-PAGE凝胶浓度3个方面优化油茶种仁双向电泳体系。结果表明:采用TCA丙酮+SDS酚抽法提取油茶种仁蛋白能够得到更多较为清晰的蛋白点;采用62 000 V·h的高压进行等电聚焦更适合油茶种仁蛋白的第一向分离;10%SDS凝胶浓度更适合油茶种仁蛋白的双向分离,试验结果为深入开展油茶种仁蛋白质组学的研究奠定基础。

Using kernel of Camellia oleifera ‘Min 43＇,a proper two-dimensional electrophoresis system for kernel of Camellia oleifera Abel. was optimized by three factors： sample preparation methods,IEF focusing program parameter settings and SDS-PAGE gel concentration. The results showed that more clear spots could be obtained by the method of TCA/acetone combined with SDS phenol extraction,the isoelectric focusing of 62 000 V·h was more suitable for the separation of protein and the concentration of 10%SDS gel was better for the separation kernel of C. oleifera protein. The result of this research laid the foundation for the following proteomics research in oil tree.