一种适用于胡杨叶片非变性蛋白质提取的方法
DOI:
作者:
作者单位:

中央民族大学,中央民族大学,中央民族大学,中央民族大学,中央民族大学,中央民族大学,中央民族大学生命与环境科学学院,生物成像与系统生物学研究中心

作者简介:

通讯作者:

中图分类号:

Q946

基金项目:

国家自然科学基金项目(面上项目,重点项目,重大项目)


A Novel Method for Nondenaturing Protein Extraction from Populus Euphratica Leaves
Author:
Affiliation:

Minzu University of China,Minzu University of China,Minzu University of China,Minzu University of China,Minzu University of China,Minzu University of China,

Fund Project:

The National Natural Science Foundation of China (General Program, Key Program, Major Research Plan)

  • 摘要
  • |
  • 图/表
  • |
  • 访问统计
  • |
  • 参考文献
  • |
  • 相似文献
  • |
  • 引证文献
  • |
  • 资源附件
  • |
  • 文章评论
    摘要:

    为探索适用于胡杨叶片非变性蛋白质的提取方法。以采自内蒙古额济纳旗天然胡杨林中胡杨成熟叶片为材料,采用咪唑法、Bis-Tris法、Tris-HCl法和新构建的Tris-SSAD法4种非变性蛋白质提取策略,分别用于提取胡杨叶片蛋白质样品,随后进行第一维非变性聚丙烯酰胺凝胶电泳(1st-DE: Native-PAGE)。相比之下,只有Tris-SSAD法获得了清晰、条带数目较多的Native-PAGE图谱,且结合第二维变性凝胶电泳(2nd-DE: SDS-PAGE)可成功分离出较多蛋白质复合体亚基或相互作用蛋白质分子,证明了Tris-SSAD法非变性温和提取特性。本研究建立并优化了适用于胡杨叶片的非变性蛋白质提取方法,为后续进一步分析蛋白质与蛋白质相互作用及蛋白质复合体功能提供了实验基础。

    Abstract:

    This study aims to establish a nondenaturing protein extracted method to study protein complexes and interacting proteins from leaves of Populus euphratica. Four approaches, including Imidazole, Bis-Tris, Tris-HCl, and new method of Tris-SSAD (Tris based Sucrose, Sodium chloride, Ascorbate, and Digitonin solution), were applied in the nondenaturing extraction of leaf proteins. Based on the first-dimensional non-denaturing gel electrophoresis (1st-DE: Native-PAGE), it was found that the highest quality protein bands on CBB stained gel were only observed in the protein extracted by the method of Tris-SSAD. Further optimized processing was also performed to obtain more perfect results. Meanwhile, the nondenatuing feature of Tris-SSAD were also successfully proven by the tandem electrophoresis, composed of nondenaturing 1st-DE (Native-PAGE) and denaturing 2nd-DE (SDS-PAGE), and a lot of protein complexes and/or interactive proteins were found in these electrophoresis gels. Finally, a new method suitable for the nondenaturing protein extraction from adverse-resistant plant leaves was achieved.

    参考文献
    相似文献
    引证文献
引用本文

韩航,马燕芳,李征珍,等. 一种适用于胡杨叶片非变性蛋白质提取的方法[J]. 科学技术与工程, 2018, 18(21): .
Han Hang, Ma Yanfang, Li Zhengzhen, et al. A Novel Method for Nondenaturing Protein Extraction from Populus Euphratica Leaves[J]. Science Technology and Engineering,2018,18(21).

复制
分享
文章指标
  • 点击次数:
  • 下载次数:
  • HTML阅读次数:
  • 引用次数:
历史
  • 收稿日期:2018-02-12
  • 最后修改日期:2018-02-12
  • 录用日期:2018-04-18
  • 在线发布日期: 2018-07-30
  • 出版日期:
×
律回春渐,新元肇启|《科学技术与工程》编辑部恭祝新岁!
亟待确认版面费归属稿件,敬请作者关注