[1]王雪,段玉玺,陈立杰.适用于大豆叶片蛋白质组分析的双向电泳最佳条件研究[J].大豆科学,2009,28(02):325-328.[doi:10.11861/j.issn.1000-9841.2009.02.0325]
　WANG Xue,DUAN Yu-xi,CHEN Li-jie.A Two-Dimensional Electrophoresis Protocol Suitable for Proteomic Analysis of Soybean Leaves[J].Soybean Science,2009,28(02):325-328.[doi:10.11861/j.issn.1000-9841.2009.02.0325]
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适用于大豆叶片蛋白质组分析的双向电泳最佳条件研究

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第28卷 期数: 2009年02期 页码: 325-328 栏目: 出版日期: 2009-04-25
Title:
A Two-Dimensional Electrophoresis Protocol Suitable for Proteomic Analysis of Soybean Leaves
文章编号:
1000-9841(2009)02-0325-04
作者:
王雪; 段玉玺; 陈立杰
沈阳农业大学，中国北方植物线虫研究所，辽宁 沈阳 110161
Author(s):
WANG Xue; DUAN Yu-xi; CHEN Li-jie
Shenyang Agricultural University，Nematology Institute of Northern China，Shenyang 110161,Liaoning,China
关键词:
大豆; 叶片; 蛋白质组; 双向电泳
Keywords:
Soybean; Leaf; proteome; Two-dimensional electrophoresiss
分类号:
S565.1
DOI:
10.11861/j.issn.1000-9841.2009.02.0325
文献标志码:
A
摘要:
针对大豆叶片中蛋白含量较低，含有大量色素、酚类、醌类等次生代谢产物干扰蛋白质双向电泳分离效果的问题，以大豆叶片为材料，对样品制备、蛋白质裂解液组分、等电聚焦（IEF）参数等条件进行研究。结果表明：采用TCA/丙酮法提取大豆叶片总蛋白，裂解缓冲液为9 molL-1尿素，2 molL-1硫脲，4%CHAPS，100 mmolL-1DTT，0.5% Bio-Lyte，IEF等电聚焦条件为24 000Vh时，2-DE图谱分离到的蛋白点效果最好。初步建立了一套适用于大豆叶片蛋白质组分析的双向电泳（2-DE）体系。
Abstract:
Proteomics is becoming a necessity in plant biology，as it is in medicine，zoology and microbiology.This study aimed at solving the problem of low content of protein in soybean leaf and the disturbance of secondary metabolites such as chromatophore，phenols and quinones to the separation of two-dimensional electrophoresis.Using leaves of soybean as material，index such as protein extraction，lysis buffer and IEF parameter were modified and improved .Results showed that，soybean leaves protein was extracted with TCA/acetone，then resolubilized in lysis buffer（9 molL-1urea，2 molL-1thiourea，100 mmolL-1DTT，0.5% Bio-Lyte），Isoelectric focusing was performed at 17℃ under the 24000Vh conditions.A 2-DE（two-dimensional electrophoresis）protocol suited for the separation of proteins from soybean leaves was elementary established.

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备注/Memo

基金项目：国家自然科学基金资助项目(30671399);农业部现代农业技术体系资助项目 （413010211-1101-01031908002）。

作者简介：王雪（1981-），女，博士研究生，从事大豆对大豆胞囊线虫的抗性研究。E-mail:wangxue813@126.com。

更新日期/Last Update: 2014-10-04