| 王 娜，谢惠民，王 川，王宏礼.冬小麦叶片抗旱蛋白质组双向电泳技术体系的优化[J].麦类作物学报,2011,31(3):443 |
| 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 |
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| DOI：10.7606/j.issn.1009-1041.2011.03.011 |
| 中文关键词: 小麦 PEG胁迫 蛋白提取方法 双向电泳 差异表达蛋白 |
| 英文关键词:Wheat PEG stress Protein extraction Two dimensional electrophoresis Differential expression protein |
| 基金项目:国家 863 计划项目(2006AA100201)。 |
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| 摘要点击次数: 2021 |
| 全文下载次数: 2550 |
| 中文摘要: |
| 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 cm、pH 4~7的IPG线性胶条，在上样量为150 μg·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)。 |
| 英文摘要: |
| 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. |
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