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当前位置: 首页 > 文章 > PEG分级法检测绿竹叶片双向电泳中的低丰度蛋白 安徽农业大学学报 2016,43 (2) 227-232
Position: Home > Articles > Polyethylene glycol fractionation improved detection of low-abundant proteins by two-dimensional electrophoresis analysis of Dendrocalamopsis oldhami proteome Journal of Anhui Agricultural University 2016,43 (2) 227-232

PEG分级法检测绿竹叶片双向电泳中的低丰度蛋白

作  者：
沈少炎;吴玉香;龙智慧;荣俊冬;何天友;陈礼光;郑郁善
单  位：
福建农林大学园林学院;福建农林大学竹类研究所
关键词：
绿竹;聚乙二醇(PEG);蛋白质提取;双向电泳;低丰度蛋白
摘  要：
蛋白质组学研究的关键问题之一是双向电泳(2-DE)中低丰度蛋白的分析,主要是因为相关高丰度蛋白的存在,导致IPG胶条无法吸胀低丰度蛋白,使得低丰度蛋白在2-DE中很难被检测出来。利用梯度浓度的PEG4000沉淀方法来富集绿竹叶片中不同丰度的蛋白质,从而使得低丰度蛋白在凝胶中显现出来。在PEG分级法和传统的全蛋白提取法的2-DE比较中,发现采用PEG分级沉淀法提取的蛋白质的数量以及类别明显增加,高丰度蛋白主要富集在8%和16%的PEG浓度组分中,使得其他组分中的低丰度蛋白与高丰度蛋白组分分别进行2-DE分析。经过对图像和数据的比较、分析,PEG分级法得到的5个组分蛋白点总数超过了1032个,大约是传统蛋白提取方法制备蛋白样品的3倍。
译  名：
Polyethylene glycol fractionation improved detection of low-abundant proteins by two-dimensional electrophoresis analysis of Dendrocalamopsis oldhami proteome
作  者：
SHEN Shaoyan;WU Yuxiang;LONG Zhihui;RONG Jundong;HE Tianyou;CHEN Liguang;ZHENG Yushan;College of Landscape, Fujian Agriculture and Forestry University;Institute of Bamboo, Fujian Agriculture and Forestry University;
关键词：
Dendrocalamopsis oldhami;;polyethylene glycol;;protein extraction;;2-DE;;low-abundant proteins
摘  要：
Poor detection of low-abundant proteins is a common problem in two-dimensional electrophoresis(2-DE) for separation of proteins in a proteome analysis. It is because a large proportion of whole-cell proteins can prevent low-abundant proteins from being uptaken by immobilized p H gradient(IPG) strip, consequently making the latter poorly detectable by 2-DE. In this work, we reported a straightforward protocol for preparation of whole-cell proteins of Dendrocalamopsis oldhami through differential polyethylene glycol(PEG) precipitation aiming at elimination of high-abundant proteins in plant protein samples. In comparison with 2-DE analysis of protein samples prepared using a conventional total protein method, a relatively high reproducibility of proteins was achieved using a PEG fractionation protocol in terms of protein quantities and protein species. The high-abundant proteins were precipitated predominantly in 8% and 16% PEG fraction. This allowed proteins of the high-abundant-containing fraction to be analyzed separately from those of other PEG fractions. After taking into account the overlapping protein spots among 2-DE gels of all fractions through image and statistical analyses, using this method, we detected a total of 1032 protein spots that were about 3 times over using the conventional total protein method. With the PEG fractionation method, low-abundance protein detection rate increased significantly.

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