小麦籽粒谷蛋白提取方法的优化与双向电泳分析  

Optimization of Extraction Method of Wheat Grain Glutenins and Two-dimensional Electrophoretic Analysis

作　　者：高璇[1] 高利艳[1] 晏月明[1] 

机构地区：[1]首都师范大学生命科学学院,北京100037

出　　处：《麦类作物学报》2008年第6期971-976,共6页Journal of Triticeae Crops

基　　金：国家自然科学基金项目(30571154;30771334);"863"项目(2006AA10Z186)

摘　　要：为了建立一套适用于小麦谷蛋白分析的双向电泳分析系统,得到更加清晰的双向电泳图谱,以高产优质小麦品种小偃6号和济麦20为材料,根据麦谷蛋白亚基含量较少的特点,对麦谷蛋白的提取方法以及提取过程中所用的试剂的种类、用量和处理时间等进行比较分析和优化,运用蛋白定量法测定提取样品的蛋白质含量,摸索出一套完整的用于双向电泳分析的麦谷蛋白提取和样品制备方法。利用此蛋白提取以及双向电泳方法可以获得分辨度较高的电泳分离图谱,特别是在低分子量谷蛋白区域,可分离出约200个蛋白点,可作为小麦籽粒蛋白质组研究的重要工具。同时,通过对不同品种和不同种子发育时期的麦谷蛋白双向电泳分离结果的分析,初步得到了麦谷蛋白在小麦籽粒生长发育过程中的合成积累情况,可为今后小麦发育蛋白质组学以及对小麦品质改良的研究提供一定的科学依据。To develop an efficient and high resolution 2-DE procedure for wheat glutenin research, in the present study, the method of glutenin subunit extraction and sample preparation for two-dimensional gel electrophoresis （2-DE） were optimized using wheat cultivars Xiaoyan 6 and Jimai 20 and by changing the type and amount of extraction reagents as well as extraction time. By using the optimized method, separation results of grain glutenins from different developmental periods in the two wheat cuhivars showed that higher resolution of the electrophoretic separation patterns could be achieved. In particular, about 200 protein dots of low molecular weight glutenin subunits were separated, suggesting that it could be used as an important tool for the studies of wheat proteome. In addition, the accumulation progress of glutenin in developing wheat embryos was obtained by the 2D gels at three development stages of two wheat cultivars （Xiaoyan 6 and Jimai 20）. The established of those methods may provid powerful tools for developmental proteomics and quality improvement of wheat in the future.

关 键 词：小麦籽粒 麦谷蛋白 双向电泳 蛋白质组 

分 类 号：Q512.4[生物学—生物化学]