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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 Platinum6.0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。
The effects of Tris-saturated phenol method and trichloroacetic acid (TCA) / acetone precipitation method on the extraction of total protein from Ganoderma lucidum fruiting bodies were compared. Two-dimensional electrophoresis patterns of the proteins from the two methods were analyzed by Image Master 2D Platinum 6.0 software, and 565 and 273 protein spots were obtained respectively. Some protein spots were smeared in the low-molecular-weight region of alkaline end by TCA / acetone precipitation , Tris-saturated phenol method can effectively remove the salt in the sample, the protein focus better, protein spots increased. The Tris-saturated phenol method can be used as the extraction method of the total protein of Ganoderma lucidum fruiting body primordium and provide a reference for the extraction of total protein of other medicinal fungi, and laid the foundation for the future differential protein expression of tripterygium wilfordii fermented with Ganoderma lucidum The foundation.