仁用杏‘围选1号’雌蕊总蛋白双向电泳体系的建立

Establishment of two-dimensional electrophoresis of apricot 'Weixuan1' pistil

为了建立一套适合仁用杏‘围选1号’雌蕊蛋白质分析的双向电泳系统,对总蛋白质的提取方法、SDS-PAGE凝胶浓度、上样量、IPG胶条梯度进行了优化。优化的条件为:酚-甲醇/醋酸铵沉淀法提取总蛋白质,浓度为12%的SDS-PAGE凝胶,24cm pH 4～7的IPG胶条,上样量100μg,采用该体系能获得图像清晰、分辨率高、重复性好的双向电泳图谱,基本满足仁用杏‘围选1号’雌蕊蛋白质的分析和研究。

In order to establish a two-dimensional gel electrophoresis （2-DE） protocol for pro- teomic study of apricot pistil, we optimized the extracting method of total protein, density ofSDS-PAGE gel,the sample loading,pH gradient of IPG strip by using apricot pistil. The opti- mized system ineludes the following steps, total proteins of sample extracted using phenol-methyl alcohol/ammonium acetate method, separating the proteins with 24 cm pH 4-7 IPGstrips, 12% SDS-PAGE, loading protein samples of 100 μg. This optimized system appeared tobe better for two-dimensional gel electrophoresis separation, which may be used as an impor- tant tool to study the apricot pistil proteome.