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水稻幼苗经缺铁胁迫诱导分别处理 1、3、5天后 ,用酚法和TCA/丙酮法提取叶片中的可溶性蛋白进行双向电泳分析 ,从而研究在缺铁条件下叶片中蛋白表达的动态变化规律。结果显示 :1 不同pHIPG胶条分离蛋白的效果不同。用pH3- 10的IPG胶条进行双向电泳 ,经考马斯亮蓝染色后 ,可在胶面上检测到大约 4 5 0个蛋白点 ,其中约有 89%的蛋白是酸性蛋白。如果用pH4 - 7的IPG胶条进行双向电泳 ,则可检测到大约 6 0 0个蛋白点 ,其中有 2 9个蛋白是上调表达 ,1个蛋白是下调表达 ,5个蛋白是诱导特异表达。 2 不同方法提取的可溶性蛋白质量不同。TCA法简单易操作 ,似乎对于碱性蛋白的抽提效果更好 ,在2 -DE图像上 ,减性端显示的蛋白点多 ;但此方法所得蛋白的再溶性差。酚法提取的蛋白再溶性好 ,所抽提的蛋白量较大 ,纯度较高。
After treated with iron deficiency stress for 1, 3 and 5 days respectively, the soluble protein in leaves was extracted by phenol method and TCA / acetone method and analyzed by two-dimensional electrophoresis to study the dynamic changes of protein expression in leaves under iron deficiency . The results showed that: 1 Different pHIPG strips of different protein separation effect. Two-dimensional electrophoresis was carried out using IP3 strips with pH 3-10. After staining with Coomassie Brilliant Blue, about 450 protein spots were detected on the plastic surface, and about 89% of the proteins were acidic proteins. Two - dimensional electrophoresis with IPG strips of pH4 - 7 detected about 60 protein spots, of which 29 were up - regulated, one was down - regulated and five were induced specific expression. 2 different methods to extract different amounts of soluble protein. TCA method is simple and easy to operate, it seems that the extraction of alkaline protein is better, in the 2-DE images, the reduced end shows more protein spots; but this method resulting protein poor re-solubility. Phenol extracted protein re-soluble, extracted a larger amount of protein, higher purity.