[1]张宁,孙晓丽,端木慧子,等.野生大豆叶片蛋白质组双向电泳技术体系的优化研究[J].大豆科学,2015,34(05):874-880.[doi:10.11861/j.issn.1000-9841.2015.05.0874]
　ZHANG Ning,SUN Xiao-li,DUANMU Hui-zi,et al.Optimization of Two-dimensional Electrophoresis Technology System for Glycine soja Leaves Proteome[J].Soybean Science,2015,34(05):874-880.[doi:10.11861/j.issn.1000-9841.2015.05.0874]
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野生大豆叶片蛋白质组双向电泳技术体系的优化研究

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第34卷 期数: 2015年05期 页码: 874-880 栏目: 出版日期: 2015-10-25
Title:
Optimization of Two-dimensional Electrophoresis Technology System for Glycine soja Leaves Proteome
作者:
张宁1; 孙晓丽2; 端木慧子1; 于洋1; 纪巍1; 朱延明1
1.东北农业大学 生命科学学院/农业生物功能基因重点实验室，黑龙江 哈尔滨150030；
2. 黑龙江八一农垦大学 农学院，黑龙江 大庆163319
Author(s):
ZHANG Ning1;  SUN Xiao-li2;  DUANMU Hui-zi1;  YU Yang1;  JI Wei1;  ZHU Yan-ming1
1.College of Life Science/Key Laboratory of Agricultural Biological Functional Genes, Northeast Agricultural University, Harbin, 150030, China;
2.Agronomy College, Heilongjiang Bayi Agricultural University, Daqing 163319, China
关键词:
野生大豆; 叶片; 蛋白质提取; 双向电泳; 蛋白组学
Keywords:
Glycine soja;  Leaves;  Protein extraction;  Two-dimensional electrophoresis;  Proteomics
DOI:
10.11861/j.issn.1000-9841.2015.05.0874
文献标志码:
A
摘要:
为给野生大豆蛋白质组学分析提供技术支撑，以耐盐碱野生大豆叶片为材料，对Tris饱和酚法、Tris-HCl法和TCA丙酮法3种蛋白质提取方法进行了比较，并对蛋白上样量、等电聚焦参数及凝胶浓度进行了优化。结果表明：使用Tri饱和酚法提取蛋白质，上样量为2 000 μg，等电聚焦90 000 V.h，凝胶浓度为10%时，可获得背景清晰、蛋白点数较多、重复性较好的蛋白双向电泳图谱。
Abstract:
To facilitate the proteomic analysis of wild soybean leaves, in this study, we established a suitable protocol for leaf protein extraction and two-dimensional electrophoresis.We extracted the total proteins of Glycine soja leaves through Tris-phenol method, Tris-HCl method and TCA-acetone method, compared the protein quantity and yield, and found that proteins extracted by Tris-phenol method displayed the highest concentration and best quality. We also compared the 2-DE images obtained by using different protein loading amounts, isoelectric focusing (IEF)parameters, and gel concentrations. Our results identified a suitable protocol for 2-DE analysis, using the Tris-phenol method for protein extraction, 2 000 μg protein loading amount, 90 000 V.h IEF condition, 10% gel concentration. Under this condition, we obtained good 2-DE images with much clearer background, more protein spots, and better reproducibility.Taken together, in this study, we establisheda suitable system for 2-DE analysis of wild soybean leaf proteins, which will lay a foundation for the following Glycine soja- leaf proteomic research.

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备注/Memo

基金项目：国家自然科学基金(31171578)；国家转基因重大专项（2011ZX08004）；黑龙江省高校科技创新团队建设计划(2011TD055)。

第一作者简介：张宁（1989-），女，硕士，主要从事植物基因工程与分子生物学研究。E-mail：zhang_ning1989@163.com。
通讯作者：朱延明（1955-），男，教授，博导，主要从事植物基因工程与分子生物学研究。E-mail：ymzhu2001@neau.edu.cn。

更新日期/Last Update: 2015-11-08