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作物学报 ›› 2012, Vol. 38 ›› Issue (05): 921-927.doi: 10.3724/SP.J.1006.2012.00921

• 研究简报 • 上一篇    下一篇

水稻籽粒蛋白双向电泳条件的优化及其蛋白组学方法的比较

李奇松，陈军，林世圣，李忠，张志兴，林文雄*   

  1. 福建农林大学生命科学学院农业生态研究所 / 福建农林大学作物生理与分子生态重点实验室, 福建福州 350002
  • 收稿日期:2011-07-08 修回日期:2011-12-19 出版日期:2012-05-12 网络出版日期:2012-03-05
  • 通讯作者: 林文雄, E-mail: lwx@fjau.edu.cn
  • 基金资助:

    本研究由国家自然科学基金项目(30871494), 教育部博士点基金(200803890006), 福建省自然科学基金(2007J0304, 2008J0042)和福建省重点学科经费资助。

Optimization of Two-dimensional Electrophoresis Condition for Rice Grain Protein and Comparison of Relevant Proteomic Methods

LI Qi-Song,CHEN Jun,LIN Shi-Sheng,LI Zhong,ZHANG Zhi-Xing,LIN Wen-Xiong*   

  1. Institute of Agricultural Ecology, School of Life Sciences, Fujian Agriculture and Forestry University / Key Laboratory of Crop Physiology and Molecular Ecology, Fujian Agriculture and Forestry University, Fuzhou 350002, China
  • Received:2011-07-08 Revised:2011-12-19 Published:2012-05-12 Published online:2012-03-05
  • Contact: 林文雄, E-mail: lwx@fjau.edu.cn

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2167

被引次数

28

摘要： 探讨适用于籽粒蛋白组学研究的策略，对深入研究籽粒的发育过程具有重要的意义。本文对比了3种不同的蛋白提取方法，优化了双向电泳中自制管胶一向的等电聚焦条件和IPG干胶条一向的等电聚焦时间，以MALDI-TOF/MS、Western-blot和磷酸化蛋白组学3个重要的蛋白质组学研究方法对胶内蛋白质鉴定分析。结果表明，可溶性蛋白提取法最适用于籽粒蛋白组学的研究; 电泳条件优化后，得到了较优的2-DE图谱; 胶内蛋白点适用于质谱分析、蛋白表达量验证(Western-blot)及籽粒蛋白的磷酸化组学的研究。本研究为下一步在蛋白组水平上分析籽粒发育提供了技术支持。

关键词: 水稻, 籽粒, 蛋白组学, 双向电泳, 蛋白磷酸化

Abstract: A suitable proteomic strategy for rice grain protein research is important for further understanding rice grain development at proteome level. In this study, three different protein extraction methods were compared and then electrophoretic conditions of hand-made gel lands and IPG lands in IEF (isoelectric focusing) were optimized. Furthermore, three important proteomic research strategies (MALDI-TOF/MS, western-blot and phosphoproteome) were used for identification and analysis of in-gel proteins. The results showed that the method of soluble protein extraction was most suitable for investigation of grain proteomics, with a well-distributed 2-DE gel profile under optimized electrophoretic conditions, and it was confirmed that MALDI-TOF/MS, western-blot and phosphoproteome were the three suitable methods for the analysis of in-gel proteins. The establishment of the suitable system for rice grain proteomic research paves the way for the next step to further analyse rice grain development at proteome level.

Key words: Rice, Grain filling, Proteomics, Two-dimensional electrophoresis, Protein phosphorylation

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