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棉花学报  2012 

棉花叶片蛋白质组双向电泳技术的优化

DOI: 1002-7807(2012)05-0468-05, PP. 468-472

Keywords: 蛋白质组,双向电泳,棉花,叶片

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Abstract:

以棉花叶片为材料,针对试材中含有大量色素、酚、醌等干扰物质的问题,从蛋白提取方法、裂解液成分、上样量等方面对棉花叶片蛋白质组双向电泳技术进行优化。结果表明采用改良的TCA/丙酮法提取棉花叶片总蛋白,含有7mol·L-1尿素、2mol·L-1硫脲、4%CHAPS、80mmol·L-1DTT、1mmol·L-1PMSF、0.3%载体两性电解质的裂解缓冲液,上样量为600μg,采用pH4~7、24cm的IPG胶条进行双向电泳时,2-DE图谱的蛋白点分布均匀、清晰且重复性好。本体系为开展棉花蛋白质组学研究奠定了技术基础。

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