[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pH3～10IPG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 a two-dimensional electrophoresis technique suitable for the research of proteome of rapeseed. [Method] With Xiangyou 17 as material, the conditions of sample preparation, gel concentration, sample loading and so on in two-dimensional electrophoresis were optimized. [Result] The optimal total protein extracted by TCA-acetone method was the best, and the obtained protein had the most points on the 2-DE map. When pH3 ~ 10IPG strips and 12% gels were used, a two-dimensional gel electrophoresis pattern with clear background, good repeatability and high resolution of protein spots was obtained when the sample volume was 250μg. [Conclusion] The study laid the foundation for the proteomics research of rape seed.