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金 艳,许海霞,徐圆圆,尚晓丽,崔党群.几种不同提取方法对小麦叶片总蛋白双向电泳的影响[J].麦类作物学报,2009,29(6):1083
几种不同提取方法对小麦叶片总蛋白双向电泳的影响
Effect of Different Extraction Protein Methods on Two dimensional Electrophoresis of Wheat Leaf Protein
  
DOI:10.7606/j.issn.1009-1041.2009.06.026
中文关键词:  小麦  叶片蛋白质  提取方法  双向电泳图谱
英文关键词:Wheat  Leaf protein  Eextraction methods  2 DE map
基金项目:国家“863”项目(2006AA10Z1F5和2006AA100102);国家科技支撑计划项目(2006BAD01A02 10)。
作者单位
金 艳,许海霞,徐圆圆,尚晓丽,崔党群 河南农业大学农学院河南郑州 450002 
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中文摘要:
      为探索小麦叶片蛋白质提取的最优方法,以普通小麦品种石4185的叶片为材料,研究了三氯乙酸/丙酮法、裂解液法、尿素/硫脲法、KCl 醋酸铵甲醇法等四种不同蛋白质提取方法对小麦叶片蛋白双向电泳图谱的影响。结果表明,三氯乙酸/丙酮法获得的蛋白点形状规则、清晰,且重复性好;裂解液法拖尾现象较轻,但蛋白点数少且欠清晰,重复性也不如前者;尿素/硫脲法的蛋白点数很少,也较为弥散,且有明显的拖尾现象;KCl 醋酸铵甲醇法使大量蛋白丢失,蛋白点数非常少,且上部竖纹干扰较重。因此,三氯乙酸/丙酮法是提取小麦叶片蛋白的较优方法。
英文摘要:
      In order to study the effect of different protein extraction methods on two dimensional electrophoresis(2 DE) and explore an optimal protein extraction method of wheat leaves, we compared the effect on 2 DE of 4 different protein extraction methods, including TCA/acetone method, lysis buffer method, urea/thiourea method and KCl ammonium acetate methanol method,using leaves of wheat cultivar Shi 4185. The results indicated that the optimal method was TCA/acetone method, which had stable, clear and repeatable protein spots. The second was lysis buffer method with slight tailing phenomena, and lower the repeatability and clarity. The 2 DE of urea/thiourea method has fewer and dispersed protein spots. The KCl ammonium acetate methanol method was the worst with more impurities and less protein spots. The results showed that the TCA/acetone method is the most suitable protein extraction method for wheat leaves .
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