|Table of Contents|

Optimization of Two-dimensional Electrophoresis Technology System for Glycine soja Leaves Proteome(PDF)

《大豆科学》[ISSN:1000-9841/CN:23-1227/S]

Issue:
2015年05期
Page:
874-880
Research Field:
Publishing date:

Info

Title:
Optimization of Two-dimensional Electrophoresis Technology System for Glycine soja Leaves Proteome
Author(s):
ZHANG Ning1 SUN Xiao-li2 DUANMU Hui-zi1 YU Yang1 JI Wei1 ZHU Yan-ming1
1.College of Life Science/Key Laboratory of Agricultural Biological Functional Genes, Northeast Agricultural University, Harbin, 150030, China;
2.Agronomy College, Heilongjiang Bayi Agricultural University, Daqing 163319, China
Keywords:
Glycine soja Leaves Protein extraction Two-dimensional electrophoresis Proteomics
PACS:
-
DOI:
10.11861/j.issn.1000-9841.2015.05.0874
Abstract:
To facilitate the proteomic analysis of wild soybean leaves, in this study, we established a suitable protocol for leaf protein extraction and two-dimensional electrophoresis.We extracted the total proteins of Glycine soja leaves through Tris-phenol method, Tris-HCl method and TCA-acetone method, compared the protein quantity and yield, and found that proteins extracted by Tris-phenol method displayed the highest concentration and best quality. We also compared the 2-DE images obtained by using different protein loading amounts, isoelectric focusing (IEF)parameters, and gel concentrations. Our results identified a suitable protocol for 2-DE analysis, using the Tris-phenol method for protein extraction, 2 000 μg protein loading amount, 90 000 V.h IEF condition, 10% gel concentration. Under this condition, we obtained good 2-DE images with much clearer background, more protein spots, and better reproducibility.Taken together, in this study, we establisheda suitable system for 2-DE analysis of wild soybean leaf proteins, which will lay a foundation for the following Glycine soja- leaf proteomic research.

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Last Update: 2015-11-08