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林业科学 ›› 2013, Vol. 49 ›› Issue (11): 189-194.doi: 10.11707/j.1001-7488.20131127

• 研究简报 • 上一篇    下一篇

PEG分级沉淀法分析天女木兰种子的低丰度蛋白

陆秀君1,3, 张晓林1,2,3, 刘广林1, 李天来2,3   

  1. 1. 沈阳农业大学林学院 沈阳 110866;
    2. 沈阳农业大学园艺学院 沈阳 110866;
    3. 沈阳农业大学设施园艺省部共建教育部重点实验室 沈阳 110866
  • 收稿日期:2012-11-16 修回日期:2013-04-07 出版日期:2013-11-25 发布日期:2013-11-26

Polyethylene Glycol Fractionation Analysis of Low-Abundant Proteins in Magnolia sieboldii Seeds

Lu Xiujun1,3, Zhang Xiaolin1,2,3, Liu Guanglin1, Li Tianlai2,3   

  1. 1. College of Forestry, Shenyang Agricultural University Shenyang 110866;
    2. College of Horticulture, Shenyang Agricultural University Shenyang 110866;
    3. Key Laboratory of Protected Horticulture of Ministry of Education, Shenyang Agricultural University Shenyang 110866
  • Received:2012-11-16 Revised:2013-04-07 Online:2013-11-25 Published:2013-11-26

关键词: 天女木兰, 种子, 蛋白质提取, PEG分级沉淀法

Abstract:

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.

Key words: Magnolia sieboldii, seed, protein extraction, polyethylene glycol fractionation

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