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Effect study of different methods to extract protein from sweet potato tuberous roots(PDF)

Chinese Journal of Applied & Environmental Biology[ISSN:1006-687X/CN:51-1482/Q]

Issue:
2014 02
Page:
211-216
Research Field:
Articles
Publishing date:

Info

Title:
Effect study of different methods to extract protein from sweet potato tuberous roots
Author(s):
YONG Bin ZHANG Yizheng FAN Wenqiao JIANG Yusong
1College of Life Science, Sichuan Normal University, Chengdu 610101, China 2Institute of Forestry and Life Sciences, Chongqing University of Arts and Sciences, Yongchuan 402168, China 3Sichuan Key Laboratory of Molecular Biology & Biotechnology, School of Life Sciences, Sichuan University, Chengdu 610064, China
Keywords:
sweet potato tuberous root two dimensional electrophoresis (2-DE) protein extraction methods SDS-PAGE proteomics
CLC:
TQ93 : S531.01
PACS:
DOI:
10.3724/SP.J.1145.2014.00211
DocumentCode:

Abstract:
In order to establish a two-dimensional electrophoresis system for the proteomics study of sweet potato tuberous roots, this study compared four protein extraction methods (homogenate method, acetone precipitation method, TCA- acetone precipitation method and phenol method). Then we evaluated these four protein preparation methods through different parameters, such as protein yield, SDS-PAGE and 2-DE techniques. In this experiment, 1.6 mg, 2.4 mg, 1.2 mg and1.9 mg soluble protein was yielded from 0.5 g fresh potato tuber using these 4 protein extraction methods, respectively. The SDS-PAGE results showed that the homogenate method and acetone precipitation method generated only few protein bands with high storage protein content (about Mr 20 × 103). By contrast, the TCA- acetone precipitation method and phenol method could separate more protein bands, with much lower band of storage protein. On 2-DE gels, the homogenization method could hardly complete the IEF, with only a few protein spots appearing in its alkaline end. The results of acetone method and TCA-acetone precipitation method were obviously better than homogenization method, but the number of protein was still less than phenol method. The number of protein spot obtained by phenol method was the most, and these protein spots were evenly distributed in the 2-DE gel. In summary, the phenol extraction method is suitable for sample preparation of sweet potato tuber for two-dimensional electrophoresis.

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Last Update: 2014-05-04