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农学学报

农学学报 ›› 2013, Vol. 3 ›› Issue (1): 1-5.

所属专题： 生物技术； 玉米

• 农艺科学 作物遗传育种 生理生化 •    下一篇

玉米叶片蛋白质双向电泳方法的优化

裴玉贺 白建芬 郭新梅 宋希云   

  • 收稿日期:2012-08-02 修回日期:2012-09-12 出版日期:2013-01-20 发布日期:2013-01-20
  • 基金资助:

    山东省农业良种工程;山东省“泰山学者岗位”（作物栽培学与育种学）

Two Dimensional Electrophoresis Methods Suitable for Maize LeafProtein Analysis

  • Received:2012-08-02 Revised:2012-09-12 Online:2013-01-20 Published:2013-01-20

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摘要：

摘要：为了建立适合玉米叶片蛋白质组研究的双向电泳方法，以抗旱玉米自交系齐319为实验材料，采用三种不同的蛋白提取方法提取玉米叶片总蛋白，进行单向SDS-PAGE和双向电泳，并进行硝酸银染色。结果表明，TCA/丙酮沉淀法适合玉米叶片蛋白质的提取。建立了适合玉米叶片可溶性蛋白的双向电泳体系，即采用24cm、pH4-7的IPG胶条，蛋白上样量0.25mg。IEF程序为:100V 1h;300V 1h;500V 1h;1000V 2h;8000V 3h;8000V 10h。SDS-PAGE的电泳条件为 1W/胶条 1h，3W/胶条，直到溴酚蓝到达玻璃底部为止。本研究为下一步在蛋白质组水平上分析干旱胁迫下玉米叶片蛋白质的变化提供了技术支持。

关键词: 马铃薯, 马铃薯, 机械化, 农艺, 栽培技术

Abstract:

Abstract: The purpose of the study is to develop 2-DE method suitable for maize leaf proteome analysis in future. Three different methods were adopted to extract leaf protein of maize line Qi319. And the prepared samples were separated by SDS-PAGE and two-dimensional electrophoresis, followed by Silver nitrate staining. The result shows that TCA/acetone precipitation method is suitable for extracting maize leaf protein. IEF method is carried out on 24cm IPG strips at pH 4-7 and protein sample weight 0.25 mg. IEF program: 100V 1 h; 300V 1h; 500 V 1 h; 1000 V 2 h; 8000 V 3 h; 8000V 10h. Condition of SDS-PAGE is 1 W/gel 1 h, 3W/gel to Bromophenol blue reaching bottom. The establishment of the suitable system for maize leaf proteomic research paves the way for the next step to further analyse maize drought-resistance at proteome level.

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