| 灵芝子实体原基双向电泳和总蛋白质提取方法的建立 |
| Two-dimensional electrophoresis and extraction methods of total protein of Ganoderma lucidum primordium |
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| 中文关键词:Tris-饱和酚法，三氯乙酸/丙酮沉淀法，药用真菌，双向性固体发酵 |
| 英文关键词:Tris-saturated phenol law, TCA/acetone precipitation method, medicinal mushroom, bi-directional solid fermentation |
| 基金项目:科技部国际合作项目（No. 2006DFA33060）；江西省科技攻关项目；南昌市科技局攻关项目 |
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| 中文摘要: |
| 比较了Tris-饱和酚法和三氯乙酸（TCA）/丙酮沉淀法对灵芝子实体原基总蛋白质的提取效果。用Image Master 2D Platinum 6.0软件分析两种方法所提蛋白质的双向电泳图谱，分别得到565和273个蛋白质点；（TCA）/丙酮沉淀法在碱性端低分子量区域有些蛋白质斑点存在拖尾现象，Tris-饱和酚法能有效去除样品中的盐分，使蛋白质的聚焦效果更好，蛋白点数增加。Tris-饱和酚法可做为灵芝子实体原基总蛋白质的提取方法并为其他药用真菌总蛋白质的提取提供参考，同时为本实验室后续研究灵芝双向性固体发酵雷公藤的蛋白差异表达奠定了基础。 |
| 英文摘要: |
| The total protein extraction effect of Ganoderma lucidum primordium by Tris-saturated phenol law and trichloroacetic acid (TCA)/acetone precipitation methods were compared. After analyzing the two-dimensional electrophoresis (2-DE) protein atlas by the software of Image Master 2D Platinum 6.0, 565 and 273 protein spots were obtained separately by the method of Tris-saturated phenol and TCA/acetone precipitation; The 2-DE gels of TCA/acetone precipitation showed strong horizontal streaks in the atlas of alkalinity end and low molecular region, while the Tris-saturated phenol law was capable of generating more protein spots, obtaining better effect of isoelectric focusing and reducing salt. Tris-saturated phenol law can be considered as a feasible protein extraction method for Ganoderma lucidum primordium and the results of this study are of great significance for developing effective protein extraction methods in other medicinal mushrooms and will lay the foundation of further study on the discrepancy expression of Ganoderma lucidum treated by bi-directional solid fermentation. |
| 刘晓云 苏明声 杨俊璋 谢小梅. 灵芝子实体原基双向电泳和总蛋白质提取方法的建立[J]. 菌物学报, 2009, 28(6): 802-805 |
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