遗传 ›› 2008, Vol. 30 ›› Issue (1): 123-126.

• 技术与方法 • 上一篇    

一种小麦高、低分子量麦谷蛋白亚基的提取方法

纪军1,2, 刘冬成2, 王静1, 李俊明1, 张爱民2   

  1. 1. 中国科学院遗传与发育生物学研究所农业资源研究中心, 石家庄 050021;
    2. 中国科学院遗传与发育生物学研究所, 北京 100101

  • 收稿日期:2007-08-01 修回日期:2007-10-10 出版日期:2008-01-10 发布日期:2008-01-10
  • 通讯作者: 张爱民

A method of extraction and separation of wheat gluten

JI Jun1,2, LIU Dong-Cheng2, WANG Jing1, LI Jun-Ming1, ZHANG Ai-Min2   

  1. 1. Center for Agricultural Resources Research, Institute of Genetics and Developmental Biology, CAS, Shijiazhuang 050021, China;
    2. National Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Acad-emy of Sciences, Beijing 100101, China
  • Received:2007-08-01 Revised:2007-10-10 Online:2008-01-10 Published:2008-01-10
  • Contact: ZHANG Ai-Min

摘要:

用7.5%的异丙醇和0.3 mol/L的NaI去除醇溶蛋白和其他单体蛋白, 以二硫苏糖醇(DTT)为强还原剂, 以4-乙烯基吡啶 (VP)保护巯基, 防止其重新氧化。在25%的异丙醇和0.04 mol/L的Tris-HCl (pH=8.0)缓冲液中提取小麦总麦谷蛋白亚基、在4%浓缩胶和13%分离胶的不连续分离体系中进行SDS-PAGE电泳, 结果表明, 该方法不仅能有效去除醇溶蛋白和其他蛋白对麦谷蛋白亚基电泳的影响, 且高分子量麦谷蛋白亚基 (HMW-GS) 和低分子量麦谷蛋白亚基 (LMW-GS)的提取分离一步完成, 更重要的是, 利用该方法提取出的HMW-GS和LMW-GS在电泳分析中, 具有高的分辨率, 可以有效区分各电泳谱带, 为进一步研究奠定了基础。

关键词: 高分子量麦谷蛋白亚基 (HMW-GS), 低分子量麦谷蛋白亚基 (LMW-GS), 提取, 分离, SDS-PAGE

Abstract:

After removing gliadin and other proteins with 7.5% 2-propanol and 0.3 mol/L NaI, the total glutenin subunits were extracted with two kinds of buffer, one containing 25% 2-propanol, 0.04 mol/L Tris-HCI (pH=8.0), 10% SDS and 2% DDT and the other containing 25% 2-propanol, 0.04 mol/L Tris-HCI (pH=8.0), 10% SDS and 1.4% VP. In SDS-PAGE electrophoresis (4% stacking gel and 13% resolving ge1) system HMW-GS and LMW-GS were clearly separated by the improved method with only a single step. The background of the SDS-PAGE gel was gliadin-free. It was more effective and applica ble to separate and distinguish the complex LMW-GS. This method will be used in wheat protein separation, variety identification and improvement.