植物学报 ›› 1999, Vol. 16 ›› Issue (02): 171-177.

• 专论与综述 • 上一篇    下一篇

木本植物蛋白提取和SDS-PACE分析方法的比较和优化

谷瑞升 刘群录 陈雪梅 蒋湘宁   

  1. 1(中国科学院植物研究所 北京 100093) 2(北京林业大学森林生物实验中心 北京 100083)
  • 出版日期:1999-03-20 发布日期:1999-03-20
  • 通讯作者: 蒋湘宁

Comparison and Optimization of the Methods on Protein Extraction and SDS-PAGE in Woody Plants

GU Rui-Sheng, LIU Qun-Lu, CHEN Xue-Mei and JIANG Xiang-Ning   

  1. 1(Institute of Botany, The Chinese Academy of Sciences, Beijing 100093) 2(Expermental Center of Forest Biology, Beijing Forest University, Beijing 100083)
  • Online:1999-03-20 Published:1999-03-20
  • Contact: JIANG Xiang-Ning

摘要: 在几种木本植物上对4种常用蛋白提取方法、4种凝胶染色和脱色方法及影响蛋白定量和SDS-PAGE电泳的因素进行了比较研究,发现:利用改良丙酮沉降法提取蛋白,不仅提取效率高,而且杂质干扰少,得到的电泳结果,蛋白条带清晰,数量多;将常规凝胶染色和脱色液中的乙醇改用为甲醇可大大改善染色和脱色效果,获得了没有背景或背景很浅的电泳结果。通过研究确定了一套优化的适用于木本植物的蛋白提取、定量、凝胶制备、电泳、染色、脱色以及干胶制备的方法。利用这一优化方法对胡杨细胞盐胁迫蛋白进行了研究,发现了与高水平盐胁迫有关的66kD蛋白和与盐胁迫和干旱胁迫均有关的28kD蛋白,获得了满意的蛋白SDS-PAGE分析结果。

Abstract: The methods of protein extraction and SDS-PAGE were compared and optimized in woody plants. The improved steps included modified protein extraction, substitution of ethanol for methanol in the solution of staining and destaining. Satisfied results were achieved in the application of the optimized method to the protein analysis of several woody plants. 66 kD protein band related to salt stress and 28 kD protein band associated with osmotic stress were obviously showed out in the analysis for the suspensivly cultured cells stressed with NaCl of Populus euphratica.