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当前位置: 首页 > 文章 > PEG分级沉淀法分析天女木兰种子的低丰度蛋白 林业科学 2013,49 (11) 189-194
Position: Home > Articles > Polyethylene Glycol Fractionation Analysis of Low-Abundant Proteins in Magnolia sieboldii Seeds Scientia Silvae Sinicae 2013,49 (11) 189-194

PEG分级沉淀法分析天女木兰种子的低丰度蛋白

作  者：
陆秀君;张晓林;刘广林;李天来
单  位：
沈阳农业大学林学院;沈阳农业大学园艺学院辽宁省设施园艺重点实验室;沈阳农业大学
关键词：
天女木兰;种子;蛋白质提取;PEG分级沉淀法
摘  要：
蛋白质组学研究的瓶颈问题之一是低丰度蛋白的检测,这些低丰度蛋白往往发挥多种重要的生理功能,如细胞防御、基因表达调节、信号传导等,因此低丰度蛋白的检测尤为重要。在双向电泳试验时,有2个因素限制了低丰度蛋白的检测:第一,IPG胶条的上样量有限;第二,高丰度蛋白的位置效应(王旭,2007)。例如,植物叶片中高丰度蛋白Rubisco含量超过50%(Gutteridge et al.,1995),这些高丰度蛋白的存在影响低丰度蛋白的分析。应用PEG分级提取法对水稻(Oryza sativa)叶片蛋白提取发现,
译  名：
Polyethylene Glycol Fractionation Analysis of Low-Abundant Proteins in Magnolia sieboldii Seeds
作  者：
Lu Xiujun;Zhang Xiaolin;Liu Guanglin;Li Tianlai;College of Forestry,Shenyang Agricultural University;Key Laboratory of Protected Horticulture of Ministry of Education,Shenyang Agricultural University;College of Horticulture,Shenyang Agricultural University;
关键词：
Magnolia sieboldii;;seed;;protein extraction;;polyethylene glycol fractionation
摘  要：
Magnolia sieboldii seeds store large amount of proteins that would interfere with determination of low abundance proteins by the electrophoresis pattern. In this study,a plant leaf protein extraction method was employed,that is,enriching high-abundance protein Rubisco by PEG. The PEG fractionation method was compared with TCA-acetone extract method in 2-DE analysis of protein samples. The result showed that the two methods had good reproducibility for extracting proteins. High-abundant proteins were precipitated predominantly in the 8% and 16% PEG fraction. With this protocol,a total 1 770 protein spots were detected,among which 50% of proteins were undetectable with the TCA-acetone method. With the PEG fractionation method,low-abundance protein detection rate increased significantly,and the PEG fractionation method,as a new method for proteomics study of M. sieboldii seeds,would be able to satisfy the follow-up test in screening the seed germination associated protein needs. The PEG fractionation method would also provide a reference for other plant seed proteins extraction.

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