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|Table of Contents|

适合杨树叶片蛋白质组学样品制备方法的比较(PDF)

《南京林业大学学报(自然科学版)》[ISSN:1000-2006/CN:32-1161/S]

Issue:
2018年02期
Page:
33-39
Column:
研究论文
publishdate:
2018-03-20

Article Info:/Info

Title:
Comparison of sample preparation methods suitable for poplar leaf proteomics
Article ID:
1000-2006(2018)02-0033-07
Author(s):
ZHEN Yan ZHENG Xiuhua SHI Jisen*
Co-innovation Center for the Sustainable Forestry in Southern China, Key Laboratory of Forest Genetics & Biotechnology of the Ministry of Education, College of Forestry, Nanjing Forestry University, Nanjing 210037, China
Keywords:
Keywords:Rubisco PEG fractionation poplar leaves two-dimensional electrophoresis(2-DE)
Classification number :
S792.11
DOI:
10.3969/j.issn.1000-2006.201706063
Document Code:
A
Abstract:
【Objective】In order to improve the detection of low abundance proteins in plant leaves, the effective extraction methods were established in poplar leaves for proteomics analysis.【Method】 Trichloroacetic acid(TCA)-acetone coupled with polyethylene glycol(PEG)fractionation and sodium dodecyl sulfate(SDS)-phenol coupled with PEG fractionation were evaluated, combined with SDS-polyacrylamide gel electrophoresis(SDS-PAGE)and two-dimensional electrophoresis(2-DE).【Result】The SDS-PAGE results showed that TCA-acetone coupled with PEG fractionation is superior to SDS-phenol coupled with the PEG fractionation method. Further, proteins with PEG fractionation and no PEG fractionation coupled with TCA-acetone were compared by using 2-DE, and the gel maps displayed that PEG fractionation is a good way to deplete the Rubiscos and improve the detection of low abundant proteins. The PEG fractionation method can detect nearly 90 more spots than nonfractionation can. 【Conclusion】LTQ-Orbitrap XL was used to identify several proteins from the PEG fractionation coupled with TCA-acetone. The TCA-acetone coupled with PEG fractionation is an effective approach for the detection of 2-DE analysis and mass spectroscopy(MS)identification of lower abundance proteins.

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Last Update: 2018-06-12