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棉花学报  2012 

棉花叶片双向电泳体系的研究

DOI: 1002-7807(2012)01-0027-08, PP. 27-34

Keywords: 棉花叶片,蛋白质提取方法,蛋白组学,双向电泳

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Abstract:

以棉花叶片为材料,比较分析了不同叶位棉花叶片总蛋白提取方法、线性固相pH梯度(ImmobilizedpHgradient,IPG)胶条pH值、上样量、水化上样缓冲液中还原剂和两性电解质等对电泳结果的影响。结果表明,三氯乙酸-丙酮沉淀法结合甲醇等有机溶剂洗涤纯化样品能获得纯度较高的棉花叶片蛋白样品。24cmpH5~8的线性IPG胶条,300μg上样量,水化上样缓冲液中用还原剂TBP(三丁基膦)代替DTT(二硫苏糖醇),两性电解质pH3~10和pH5~8按1∶1(V/V)的比例混合使用,并结合适宜的双向电泳参数,可获得背景清晰、重复性较好、蛋白点数目多且分辨率高的双向电泳图谱。从而建立了一套适合于棉花叶片蛋白组学分析的双向电泳技术体系。

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