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以甘蓝型油菜温敏核不育系SP2S及其近等基因系SP2F为材料,分别采用TCA/丙酮法和Tris-丙酮-酚法提取花蕾总蛋白,并优化电泳体系,对可育和不育花蕾中的蛋白质表达进行差异分析。结果表明:TCA/丙酮法适合于花蕾总蛋白的提取,采用10%浓度的分离胶,上样量为450μg,用pH 4~7的17 cm线性胶条和改良的考马斯亮蓝染色法得到了背景清晰、蛋白点分布均匀且重复性好的电泳凝胶图谱。利用这一实验体系,筛选出SP2S和SP2F之间大量的差异蛋白点。相比SP2F,仅在SP2S中表达的有13个点,上调2倍以上的点有4个,下调2倍以上的点有9个。
The thermo-sensitive genic male sterile line SP2S and its near isogenic line SP2F were used as materials to extract the total bud bud protein by TCA / acetone method and Tris-acetone-phenol method respectively, and the electrophoresis system was optimized for fertile and sterile Differences in protein expression in flower buds were analyzed. The results showed that the TCA / acetone method was suitable for the extraction of total bud protein. A 10% concentration gel was used, the sample volume was 450μg, and the 17cm linear gel with pH 4-7 and the modified Coomassie brilliant blue staining Clear background, uniform distribution of protein spots and good repeatability gel electrophoresis. Using this experimental system, a large number of differentially expressed protein spots between SP2S and SP2F were screened out. Compared to SP2F, there were only 13 spots expressed in SP2S, 4 spots increased more than 2-fold, and 9 spots decreased more than 2-fold.