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鹿尾腺总蛋白双向电泳体系的建立

Keywords: 鹿尾,鹿尾腺,双向电泳,蛋白质组学,不同提取方法

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Abstract:

目的:建立一套适用于鹿尾腺总蛋白分析的双向电泳体系,为实现通过蛋白质组学研究手段探究鹿尾腺的组织发育及其发挥药理作用的机制奠定基础。方法:使用三氯乙酸(TCA)/丙酮沉淀法、饱和酚抽提法和Trizol法3种方法提取鹿尾腺中总蛋白,采用固相pH梯度胶条进行双向电泳,并对蛋白上样量和等电聚焦参数进行选择和优化,凝胶考马斯亮蓝染色后用PDQuest8.0软件进行图谱分析。结果:Trizol法得到的总蛋白质纯度最高,能满足双向电泳分析对样品的要求,蛋白质样品在7cmpH3~10IPG胶条上,上样量为300μg,按优化的程序Ⅱ进行双向电泳,可以得到较满意的双向电泳图谱,此时清晰可见蛋白点数多达209个。结论:通过优化双向电泳条件,建立了适合鹿尾腺蛋白质组学研究的双向电泳体系。同时该体系可以为其他类似的动物组织样品制备和双向电泳提供借鉴。

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