| 金心黄杨（Euanymus japonica L. cv. aureopictus ）叶片总蛋白质提取及双向电泳技术 |
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| 引用本文： | 郝强,;葛秀秀,;田宇,;颜粤,;关雪莲.金心黄杨（Euanymus japonica L. cv. aureopictus ）叶片总蛋白质提取及双向电泳技术[J].北京农学院学报,2009(4):1-4. |
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| 作者姓名： | 郝强 ;葛秀秀 ;田宇 ;颜粤 ;关雪莲 |
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| 作者单位： | [1]北京农学院园林系,北京102206; [2]北京农学院生物技术系,北京102206 |
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| 基金项目： | 北京市教委科技面上项目（km200810020004,km20031002008） |
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| 摘 要： | 通过比较2种不同的植物叶片总蛋白质提取方法（三氯乙酸／丙酮法和酚法）和优化双向电泳各试验环节，建立金心黄杨叶片总蛋白质的提取方法，以及可以对其蛋白质组进行质谱分析的双向电泳条件。结果采用优化后的酚法进行金心黄杨叶片总蛋白质的提取，IEF蛋白上样量150μg，SDS-PAGE采用12．5％T凝胶，电泳结束后用考马斯亮蓝G-250染色，Melanie 7．0软件分析后得到约274个可分辨蛋白点。
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| 关 键 词： | 双向电泳 叶片蛋白提取 金心黄杨 植物蛋白质组学 |
Establishment of Extraction Method and 2-Dimensional Eiectrophoresis Conditions for Leaves Proteome Analysis of Euonymus japonica L. cv. aureopictus |
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| Institution: | HAO Qiang , GE Xiu-xiu , TIAN Yu , YAN Yue , GUAN Xue-lian . (a. Department of Landscape Design and Forestry, h. Department of Biotechnology, Beijing University of Agriculture, Beijing 102206,China) |
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| Abstract: | By comparing two different sample preparation methods （TCA/acetone and phenol） and optimizing the protein extraction procedure and the electrophoresis conditions, A phenol extraction method of total proteins from Euonymusjaponica cv. aureo-pictus leaves was established. As a result ,the first-dimensional electrophoresis was immobilized pH gradients isoelectric focusing ,150μg protein/7cm strip was added, and second dimensional etectrophoresis was vertical electrophoresis of 12.5 % T SDS-PAGE. With Coomassie Brilliant Blue G-250 stainning , more than 274 protein spots were detected on the gels of phenol extracts. These results provided the foundation for further proteomic analysis of evergreen hoard-leaf plant under cold stress. |
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| Keywords: | 2-DE leaf protein extraction Euonymus japonica L cv a ureo-pictus plant proteomics |
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