| PEG分级法检测绿竹叶片双向电泳中的低丰度蛋白 |
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| 引用本文： | 沈少炎,吴玉香,龙智慧,荣俊冬,何天友,陈礼光,郑郁善.PEG分级法检测绿竹叶片双向电泳中的低丰度蛋白[J].安徽农业大学学报,2016,43(2):227-232. |
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| 作者姓名： | 沈少炎 吴玉香 龙智慧 荣俊冬 何天友 陈礼光 郑郁善 |
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| 作者单位： | 福建农林大学园林学院,福州 350002;福建农林大学竹类研究所,福州 350002;福建农林大学竹类研究所,福州,350002 |
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| 基金项目： | 福建省科技重大专项（2013NZ0001）和福建农林大学校重点建设项目专项（6112C039D）共同资助。 |
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| 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。
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| 关 键 词： | 绿竹 聚乙二醇(PEG) 蛋白质提取 双向电泳 低丰度蛋白 |
| 收稿时间： | 2015/10/23 0:00:00 |
Polyethylene glycol fractionation improved detection of low-abundant proteins by two-dimensional electrophoresis analysis of Dendrocalamopsis oldhami proteome |
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| SHEN Shaoyan,WU Yuxiang,LONG Zhihui,RONG Jundong,HE Tianyou,CHEN Liguang and ZHENG Yushan.Polyethylene glycol fractionation improved detection of low-abundant proteins by two-dimensional electrophoresis analysis of Dendrocalamopsis oldhami proteome[J].Journal of Anhui Agricultural University,2016,43(2):227-232. |
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| Authors: | SHEN Shaoyan WU Yuxiang LONG Zhihui RONG Jundong HE Tianyou CHEN Liguang and ZHENG Yushan |
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| Institution: | College of Landscape, Fujian Agriculture and Forestry University, Fuzhou 350002; Institute of Bamboo, Fujian Agriculture and Forestry University, Fuzhou 350002,College of Landscape, Fujian Agriculture and Forestry University, Fuzhou 350002; Institute of Bamboo, Fujian Agriculture and Forestry University, Fuzhou 350002,College of Landscape, Fujian Agriculture and Forestry University, Fuzhou 350002; Institute of Bamboo, Fujian Agriculture and Forestry University, Fuzhou 350002,Institute of Bamboo, Fujian Agriculture and Forestry University, Fuzhou 350002,College of Landscape, Fujian Agriculture and Forestry University, Fuzhou 350002; Institute of Bamboo, Fujian Agriculture and Forestry University, Fuzhou 350002,Institute of Bamboo, Fujian Agriculture and Forestry University, Fuzhou 350002 and College of Landscape, Fujian Agriculture and Forestry University, Fuzhou 350002; Institute of Bamboo, Fujian Agriculture and Forestry University, Fuzhou 350002 |
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| Abstract: | 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 pH 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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| Keywords: | Dendrocalamopsis oldhami polyethylene glycol protein extraction 2-DE low-abundant proteins |
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