梯度胶对人参蛋白双向电泳分离效果的影响  

Effects on Two-Dimensional Electrophoresis Seperation of Panax Ginseng Proteins by Gradient Polyacrylamide Gels

作　　者：孙立伟[1] 雷秀娟[2] 麻锐[1] 陈佳庆[1] 姜锐[1] 赵大庆[3] 

机构地区：[1]北华大学生命科学中心,吉林吉林132013 [2]中国农业科学院特产研究所,吉林吉林132109 [3]长春中医药大学,长春130117

出　　处：《光子学报》2010年第12期2118-2122,共5页Acta Photonica Sinica

基　　金：国家科技部支撑计划项目(2007BAI38B02);吉林市科技发展计划项目(200813)资助Tel:0432-64608354

摘　　要：为了建立高质量、高分辨率人参蛋白双向电泳图谱来对人参蛋白进行功能分析与研究,提出一种利用双向梯度胶电泳技术.第一向等电聚焦电泳分离后,第二向分别采用12.5%浓度的均一胶和10%～12.5%、12.5%～15%、10%～15%三种不同浓度的SDS-PAGE梯度胶来获取人参蛋白双向电泳图谱,并利用Image Scanner和Image Master 2D Platinum version6.0对图像进行扫描及比较分析.结果表明,采用10%～15%浓度梯度胶获得双向电泳图谱中蛋白点圆滑、形状规则,分离效果好;有效地减少了人参中高丰度蛋白对中低丰度蛋白的掩盖,Mw>75 kDa的可检测到的低丰度和大分子量蛋白点数明显增加.研究结果为高质量人参蛋白表达图谱的建立奠定了良好的实验基础,同时也为其它五加科植物蛋白质组学研究工作的开展提供了方法学上的指导.In order to promote the research and analysis of panax ginseng proteins through the high quality and resolution 2-DE maps of panax ginseng,a two dimensional electrophoresis technology by gradient polyacrylamide gels is presented.In this work,total sample proteins are firstly extracted from the ginseng root,and then are seperated by IEF and 12.5% humogeneous polyacrylamide gel and 10%-12.5%,12.5%-15%,10%-15% gradient polyacrylamide gels.After all images are scaned and analyzed by Image Scanner and Image Master 2D Platinum version 6.0 system,the optional concentration of gradient polyacrylamide gel is obtained.A lot of low abundance protein spots and large molecular weight spots are observed in 10%-15% gradient polyacrylamide gel map,which indicate that the seperation efficiency of gradient gel is highly improved.The results establish the fundation of high quality 2-DE maps of panax ginseng,andprovide a tool for the proteomic analysis of other Araliaceae plants.

关 键 词：梯度胶 人参蛋白 双向电泳 

分 类 号：Q816[生物学—生物工程]