适用于小麦叶片蛋白质组分析的样品制备方法  

Sample Preparation Methods Suitable for Wheat Leaf Proteome Analysis

作　　者：刘伟霞[1] 潘映红[1] 

机构地区：[1]中国农业科学院作物科学研究所/国家农作物基因资源与基因改良重大科学工程,北京100081

出　　处：《中国农业科学》2007年第10期2169-2176,共8页Scientia Agricultura Sinica

基　　金：国家自然科学基金资助项目(30471060)

摘　　要：【目的】建立适用于小麦叶片蛋白质组分析的样品制备方法。【方法】以Taichung29小麦苗期叶片为材料,分别采用改进的酚提取-甲醇/醋酸铵沉淀法及尿素/硫脲提取法提取总蛋白,进行双向电泳分离和胶体考染,并选取代表性的蛋白点进行MALDI-TOF质谱分析及数据库搜索。【结果】上述的两种方法可检测到较多的蛋白点数,分别为1016±203和1014±281,并能成功地完成代表性蛋白点的鉴定。【结论】作者建议,进行小麦叶片蛋白质组分析时,可采用本实验提供的两种样品制备方法。[Objective] The purpose of the study is to develop sample preparation methods suitable for wheat leaf proteome analysis in future. [Method ] Two improved methods, phenol extraction-methanol/ammonium acetate precipitation and urea/thiourea extraction, were adopted to extract leaf protein of wheat line Taichung 29. The prepared samples were separated by two-dimensional electrophoresis, followed by colloidal Coomassie Blue G-250 staining. Some protein spots were selected for MALDI-TOF MS analysis and database searching. [Result] The result shows that by using the two methods, 1 016±203 and 1 014±281 spots could be detected respectively on 2-DE gels, and the selected protein spots could be efficiently identified. [Conclusion] The authors suggest that the two sample preparation methods provided here can be used in wheat leaf proteome analysis.

关 键 词：小麦叶片 样品制备 双向电泳 质谱分析 蛋白质组 

分 类 号：S512.1[农业科学—作物学]