[an error occurred while processing this directive]

Plant Diversity ›› 2009, Vol. 31 ›› Issue (04): 357-362.DOI: 10.3724 SP.J.1143.2009.09043

• 研究论文 • 上一篇    下一篇


杨 君1 , 杨云强2, 杨永平1 , 胡向阳1   

  1. 1 中国科学院昆明植物研究所中国科学院青藏高原研究所昆明部, 云南昆明 650204;
    2 黑龙江八一农垦大学生命科学技术学院, 黑龙江大庆 163319
  • 收稿日期:2009-03-09 出版日期:2009-08-25 发布日期:2009-08-25
  • 通讯作者: 胡向阳

The Sample Preparation and the Improvement Method of Two-Dimensional Electrophoresis of Proteins from Antiaris toxicaria (Moraceae) Seed

YANG Jun1 , YANG Yun-Qiang2, YANG Yong-Ping1 , HU Xiang-Yang1   

  1. 1 Kunming Institute of Botany, Chinese Academy of Sciences, Kunming 650204 , China;
    2 Heilongjiang Bayi Agricultural University , College of Life Science , Daqing 163319 , China
  • Received:2009-03-09 Online:2009-08-25 Published:2009-08-25
  • Contact: HU Xiang-Yang

摘要: 建立箭毒木( Antiaris toxicaria) 种子总蛋白的提取方法, 以及可以对其蛋白质组进行分析的双向电泳条件。通过各种条件的优化与组合, 建立了以TCA-丙酮为基础的Tris-HCl 提取法提取总蛋白, 第1 向电泳为固相pH 梯度等电聚焦, 第2 向电泳为垂直平板SDS-PAGE 的双向电泳体系。通过对样品制备、样品溶解、等电聚焦电泳、SDS———聚丙烯酰胺凝胶电泳以及染色方法等关键步骤进行分析, 获得了满意的双向电泳图谱。在探索适合箭毒木种子蛋白质组学研究双向电泳方法中, 比较了三氯乙酸- 丙酮沉淀法、和Tris-HCl 法, 以及对双向电泳过程中的关键步骤的改良, 认为Tris-HCl 法为最适方法, 所得图谱背景清晰,蛋白质信息量最大, 为箭毒木属植物的差异蛋白质组学的后续研究打下了坚实的基础。

关键词: 箭毒木, 样品制备, 双向电泳

Abstract: We have established the extraction method of the total proteins of Antiaris toxicaria seed and the appropriate conditions for the two-dimensional eiectrophoresis (2-DE) of Antiaris toxicaria seed proteome . Through integrating and optimizing various conditions, we also established the 2-DE system including the sample preparation method with Tris-HCL based on TCA-actone , and the first-dimensional electrophoresis with immobilized pH gradients isoelectric focusing and the second dimensional electrophoresis with vertical electrophoresis of SDS-PAGE. We have gained a good 2-DE map in the analysis of sample preparation、sample resolution、isoelectric focusing ( IEF) 、SDS-PAGE and gel staining. In order to explore the extraction method of proteome and technique of 2-DE , we compared the extraction method between the TCA- actone and the Tris-HCl , and improved the key steps of 2-DE . The Tris-HCl extraction was the most appropriate for Antiaris toxicaria seed proteome analysis because of the highest resolution and more informative 2-DE map . Those were the solid basis for the later research of studying on the discrepancy expression proteomics of Antiaris toxicaria Seed .

Key words: Antiaris toxicaria