大豆科学2016，Vol.35Issue(5)：810-817,8.DOI:10.11861/j.issn.1000-9841.2016.05.0810
大豆种子蛋白质组样品制备方法研究
A Study on Sample Preparations for Proteomics of Soybean Seeds
摘要
Abstract
By using pheno meth anolammonium acetate,TCA acetone or urea thiourea methods,seed proteins of soybean Zhonghuang 13 were extracted at first and second step.In order to explore the efficient methods of preparing sample for proteomics of soybean seeds.The protein extraction obtained by urea thiourea method at first step and the protein extraction obtained by pheno meth anolammonium acetate method at second step,and the first step extracted protein,which was separated further by one-dimensional electrophoresis,were analyzed with nano-LC-MS/MS respectively.The results showed that the protein yield of first and second extractions obtained by urea thiourea method and following by pheno meth anolammonium acetate method reached 29.18％ and 4.08％ respectively,and was the highest among extractions obtained by other combined methods.A total of 2 246 protein groups was identified from three replicate samples of first and second extraction,and there were significant differences between them.Average 1 167 protein groups were identified from the sample extracted by urea thiourea,and average 1 868 protein groups from the urea thiourea extracted sample which were separated further by one-dimensional electrophoresis.The results indicated that the effective sample preparation and pre-separation could enhance the yields and identification of the protein from soybean seeds.关键词
大豆种子/蛋白质组学/样品制备/纳升级液相色谱串联质谱Key words
Soybean seed/Proteomics/Sample preparation/Nano LC-MS/MS分类
农业科技引用本文复制引用
林杨杰,赵明,杨生超,潘映红..大豆种子蛋白质组样品制备方法研究[J].大豆科学,2016,35(5):810-817,8.基金项目
中国农业科学院科技创新工程(作物分子标记技术及其应用科研创新团队). （作物分子标记技术及其应用科研创新团队）
