| 周丽敏，马文洁，武炳瑾，张德强，杨智全，孙道杰.小麦穗突变体 Sda1与其野生型叶片总蛋白双向电泳体系的建立[J].麦类作物学报,2016,36(12):1623 |
| 小麦穗突变体 Sda1与其野生型叶片总蛋白双向电泳体系的建立 |
| Establishment of Two-Dimensional Electrophoresis System for the Total Leaf Proteins Separation in the Spike Mutant Sda1 and Its Wild-Type |
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| DOI：10.7606/j.issn.1009-1041.2016.12.11 |
| 中文关键词: 小麦 穗突变体 叶片 双向电泳 2-DE图谱 |
| 英文关键词:Wheat Spike mutants Leaf Two-dimensional electrophoresis 2-DE map |
| 基金项目:国家重点基础研究计划项目（2014CB138100）；陕西省自然科学基金项目（2015JM3094）；陕西省重点科技创新团队项目（2014KCT-25） |
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| 中文摘要: |
| 为探索适合小麦穗突变体 Sda1叶片蛋白的双向电泳体系，寻找Sda1与其野生型叶片的差异蛋白，以Sda1与其野生型的抽穗期叶片为材料，从蛋白质提取、双向电泳条件等方面对适合 Sda1及其野生型叶片蛋白的双向电泳体系进行探索。分析试验得到的双向电泳图谱，发现利用TCA/丙酮提取叶片总蛋白，用pH 4~7的17 cm线性胶条，采用13%浓度的分离胶，上样量为900 μg，利用改良的等电聚焦程序，能够得到背景清晰、分辨率高的电泳图谱；电泳图谱在低分子区蛋白点分布均匀，并且重复性好。利用PDQuest 8.0.1 软件分析图谱，得到27个差异点蛋白，相比于野生型植株， Sda1突变体表现为蛋白表达量下调与缺失，发现6个缺失蛋白，21个表达下调蛋白。 |
| 英文摘要: |
| In order to explore suitable 2-DE(two-dimensional electrophoresis) system for Sda1(spike development atrophy 1) leaf protein to screen differential protein expression between Sda1 and its wildlife type in wheat(Triticum).The protein extraction method and two-dimensional electrophoresis conditions were studied. A clear background, well distributed protein spots and reproducible 2-DE maps can be obtained by using the TCA/acetone protein extraction method, with 13% separation gel, a sample amount of 900 μg, the linear strip 17 cm immobilized pH gradient(IPG) strip pH 4 to 7, modified isoelectric focusing(IEF) process, and modified Coomassie Brilliant Blue for staining. Using this optimized 2-DE system, some differential protein spots between Sda1 and its wildlife-type were found. Compared with its wildlife-type,21 spots were down-expressed, and six spots were not expressed in Sda1. |
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