菜籽蛋白质组双向电泳技术体系的优化研究

摘要

[目的]建立适用于菜籽蛋白质组研究的双向电泳技术.[方法]以湘油17号为材料,对双向电泳技术中的样品制备方法、凝胶浓度、上样量等条件进行了优化.[结果]使用TCA-丙酮法提取菜籽总蛋白最佳,所获蛋白在2-DE图谱上获得的点数最多.当采用pH 3-10 IPG胶条和12％的凝胶、上样量为250μg时,可得到背景清晰、重复性好、蛋白点分辨率较高的双向电泳图谱.[结论]为开展油菜种子蛋白质组学研究奠定了基础.%[Objective] The research aimed to establish the two-dimensional electrophoresis (2-DE) technology which wes suitable for the rapeseed proteome research. [Method] Xiangyou 17 was as the material. The sample preparation method, gel concentration and loading amount,etc. in 2-DE technology were optimized. [Result] The best extraction method of total protein of rapeseed was TCA-acetone method, and the protein spots on 2-DE map were the most. When lPG strip (pH 3 -10) and 12% gel were used, and the loading amount was 250 μg, the twodimensional electrophoresis map with the clear background, good repeatability and high protein spot resolution was obtained. [Concusion] The research laid the foundation for carrying out the rapeseed proteomics research.