黄瓜叶片蛋白质双向电泳样品分级优化  

Improvement of Cucumber Leaf Protein Preparation Method for 2-DE

作　　者：任丽萍[1] 范海延[1] 王珊珊[1] 郝宇涵[1] 宫玉洁[1] 朱延姝[1] 

机构地区：[1]沈阳农业大学生物科学技术学院,沈阳110866

出　　处：《西北植物学报》2011年第8期1706-1710,共5页Acta Botanica Boreali-Occidentalia Sinica

基　　金：国家自然科学基金(30700542);辽宁省教育厅课题(2008624);沈阳市科学技术计划基金(F10-205-1-04)

摘　　要：以‘津研4号’苗期叶片为材料,采用聚乙二醇(PEG)分级沉淀法对黄瓜叶片蛋白质样品进行分级分离,将黄瓜叶片中存在的高丰度蛋白1,5-二磷酸核酮糖羧化酶/加氧酶特异性地集中于一个组分之中,以提高对黄瓜叶片蛋白质双向电泳中低丰度蛋白质的检测率。结果表明:(1)分级后各组分和全蛋白在SDS-PAGE谱带中差异显著,全蛋白得到了有效的分离。(2)二维电泳图谱上点的分辨率有很大的提高,所有组分的点数是未分级前的3倍之多。(3)浓度为24%的PEG-4000富集高丰度蛋白Rubisco的效果最好。该方法可推广应用于黄瓜叶片蛋白质组分析的样品制备。In order to improve the detection rate of cucumber leaf low-abundance proteins for 2-DE,we report a straightforward protocol for preparation of whole-cell proteins through differential polyethylene glycol（PEG） precipitation aiming at separation of Rubisco from plant protein samples by using the seedling leaves of cucumber cultivar,Jinyan 4.The result showed all the PEG fractions and the whole-cell proteins had significant differences in the SDS-PAGE,which indicated the whole-cell protein were effectively separated.Rubisco were precipitated predominantly in the 24% PEG fraction,which improved the detection rate of low-abundance proteins.A new method for the proteomics study of cucumber is provided based on 2-DE.

关 键 词：黄瓜叶片 1 5-二磷酸核酮糖羧化酶/加氧酶 聚乙二醇 双向电泳 

分 类 号：Q503[生物学—生物化学]