摘要
蛋白质组技术是植物基因功能鉴定和分析的强有力工具。经典的双向电泳技术是蛋白质组研究的核心技术,但差异蛋白质组分析使其具有更大的挑战性,因此在蛋白质水平的基因表达的定量分析研究需要更高灵敏度和更宽动力线性范围的技术。双向差异凝胶电泳(two dimension difference gel electrophoresis,2D-DIGE)是一种新出现的荧光标记的定量蛋白质组学技术,比经典的2-DE具有更高的动力学范围和灵敏性。该方法通过引入内标使得多个样品在一块胶上分离,进而减少了实验条件不一致引起的误差。2D-DIGE结合质谱、生物信息学及高可信度的统计分析使得成功地定量、鉴定植物蛋白质成为可能。本文综述了2D-DIGE蛋白质组学的基本原理,实验方法,2D-DIGE蛋白质组的优缺点和可能解决的方法,以及该技术体系在植物研究中的应用。
Proteomics is powerful tool for characterization gene function of plants. Classical two-dimensional gel electrophoresis （2-DE） is a central tool ofproteome research, however, is challenging in analysis of comparative proteome. Quantification of gene expression at the protein level requires sensitive visualization and wide linear range technique. Difference gel electrophoresis 2-DE is novel tool for quantitative proteomics, and has more dynamics range and sensitivity compared with traditional gel electrophoresis. With incorporation of internal stan- dard, multiple samples were separated on a gel, minimizing the techniques variation. 2D-DIGE, mass spectrometry and high statistical confidence enable the successful identification of plant protein. The article reviews the basic principle of based-DIGE proteomics, operational procedure including sample preparation, experimental design, imaging and statistical analysis. The advantages and disadvantages of 2D-DIGE, possible application in plant research and the solution to avoid the limitation were also discussed.
出处
《分子植物育种》
CAS
CSCD
2008年第2期405-412,共8页
Molecular Plant Breeding
基金
国家林业局948创新项目
江苏省333人才工程项目
江苏省博士后基金项目
中国博士后基金项目资助
