冬小麦叶片抗旱蛋白质组双向电泳技术体系的优化  

Winter Wheat Leaf Drought Protein and Two-dimensional Electrophoresis System Optimization

作　　者：王娜[1] 谢惠民[1] 王川[1] 王宏礼[1] 

机构地区：[1]西北农林科技大学农学院,陕西杨凌712100

出　　处：《麦类作物学报》2011年第3期443-449,共7页Journal of Triticeae Crops

基　　金：国家863计划项目(2006AA100201)

摘　　要：为给小麦抗旱蛋白质组学研究提供技术支撑,以两个抗旱性不同的冬小麦品种周麦18和晋麦47三叶期幼苗为材料,比较分析了PEG胁迫处理后两种不同蛋白质提取方法TCA/丙酮法和磷酸钠缓冲液研磨法对IEF/SDS-PAGE双向电泳的影响,并在IPG胶条pH范围、蛋白质上样量和SDS-PAGE胶浓度等方面进行了探索与优化。结果表明,采用TCA/丙酮法提取叶片全蛋白,选用17 cmp、H4~7的IPG线性胶条,在上样量为150μg.胶条-1,以及12%SDS-PAGE胶浓度下进行双向凝胶电泳,蛋白质能够更好地被分离,2-DE图谱上可分辨出约400个蛋白点。利用该体系比较了PEG胁迫后两个品种双向电泳图谱的差异,周麦18有9个蛋白点存在差异,其中5个下调表达,4个上调表达（3个新诱导表达）;晋麦47有5个蛋白点存在差异,均上调表达（1个新诱导表达）。The effects of two different protein extraction method（TCA/acetone precipitation and sodium phosphate buffer grinding method） on two-dimensional electrophoresis technology were compared,and the pH range of IPG gel strip,the loading quantity of samples and the gel concentrations were also optimized by PEG-treated on the three leaf period of two different drought-resistant winter wheat of Jinmai 47 and Zhoumai 18.The result showed that the protein were separated obviously by taking the TCA/acetone precipitation with pH 4~7,17 cm IPG line strip,150 μg loading quantity and 12% concentrations of SDS-PAGE gel,and total 400 protein spots were detected on 2-DE map.Nine protein spots in Zhoumai 18 were present difference in the protein map,including five down-regulated and four up-regulated（three novel induced expression）;five up-related present in Jinmai 47,including one novel induced expression.

关 键 词：小麦 PEG胁迫 蛋白提取方法 双向电泳 差异表达蛋白 

分 类 号：S512.11[农业科学—作物学] S312