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

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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

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

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