适用于毛白杨芽双向电泳分析的蛋白质提取方法  

Optimal protein extracting methods of Populus tomentosa buds for two-dimensional gel electrophoresis

作　　者：谢进[1] 田晓明[1] 刘淑欣[1] 侯佳音[1] 盖颖[1] 蒋湘宁[1,2] 

机构地区：[1]北京林业大学生物科学与技术学院 [2]国家林业局树木花卉育种与生物工程实验室

出　　处：《北京林业大学学报》2013年第4期144-148,共5页Journal of Beijing Forestry University

基　　金："863"国家高技术研究发展计划项目(2011AA100203);国家自然科学基金项目(NSF30630053)

摘　　要：本实验以毛白杨芽为实验材料,采用Tris-酚法、三氯乙酸-丙酮法和三氯乙酸-丙酮+Tris-酚法对其蛋白质提取方法进行比较,采用双向电泳分离蛋白质,通过PDQuest软件和质谱对这3种提取方法得到的双向电泳胶进行分析。结果显示,三氯乙酸-丙酮法得到的蛋白点数量为231±4,得到的鲜样中的总蛋白质含量为(3.57±0.618)mg/g,均高于其他两种提取方法,并且所得到的双向电泳胶更清晰,为毛白杨芽蛋白质提取的最适方法。Three protein extracting methods （Tris-phenol method, TCA/acetone method and TCA/acetone ± Tris-phenol method） were compared with the materials of P. tomentosa buds and described in this report. To determine which protein extracting method is most suitable for P. tomentosa buds, two-dimensional electrophoresis was applied to the separation of proteins, PDQuest software and mass spectrometry were used to characterize the protein spots from 2D-gel. Among the three protein extracting methods, TCA/ acetone method was the most suitable one for P. tomentosa buds and yielded （3.57 ± 0. 618） mg/g total protein amount in fresh sample, 231 ± 4 protein spots on the 2D-Gel and the clearest 2D-Gel images, which were higher in total amount, more in spot number and clearer in 2D-Gel image than other two methods.

关 键 词：毛白杨芽 蛋白质提取 双向电泳 质谱 

分 类 号：S718.46[农业科学—林学]