论文部分内容阅读
通过对IPG胶条梯度、上样方式、上样量、聚焦条件等4方面条件的优化,建立了番茄子叶总蛋白双向电泳体系,即采用TCA丙酮沉淀法提取番茄子叶总蛋白,选用24cmpH3-7NL的IPG胶条,采用水化上样,上样量300μg,按聚焦程序Ⅰ或Ⅱ聚焦后进行双向电泳分离和银染染色。若采用制备胶,以获取高含量蛋白点,则将上样量提高至1.5mg,按聚焦程序Ⅲ进行聚焦后进行双向分离,并采用胶体考马斯亮蓝染色。采用该优化的体系可以获得分辨率高、重复性好的双向电泳图谱。
Through the optimization of four conditions of IPG strip gradient, sample loading, sample loading and focusing conditions, a two-dimensional electrophoresis system of total protein of cotyledon of tomato was established. TCA acetone precipitation method was used to extract the total protein of tomato cotyledons. 24cmpH3-7NL IPG strips, using hydration samples, the amount of 300μg, according to the focusing program Ⅰ or Ⅱ focus after two-dimensional electrophoresis separation and silver staining. If the preparation of plastic to obtain high protein spots, the sample will be increased to 1.5mg, according to the focusing program Ⅲ focus after two-way separation, and colloidal Coomassie blue staining. The optimized system can obtain two-dimensional gel electrophoresis with high resolution and good repeatability.