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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]

2014 02
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Effect study of different methods to extract protein from sweet potato tuberous roots
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
sweet potato tuberous root two dimensional electrophoresis (2-DE) protein extraction methods SDS-PAGE proteomics
TQ93 : S531.01

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.


1 陶向, 张勇为, 姜玉松, 王海燕, 张义正. 甘薯块根储藏过程中的淀粉含量变化. 应用与环境生物学报, 2010, 16: 741-744 [Tao X, Zhang YW, Tong Y. Changes in root starch contents of sweet potato cultivars during storage. Chin J Appl Environ Biol, 2010, 16: 741-744]
2 陶向, 童英, 张勇为, 姜玉松, 王海燕, 张义正. 甘薯鲜薯不同储存期呼吸速率的测定与分析. 四川大学学报(自然科学版), 2010, 47 (5): 1131-1136 [Tao X, Tong T, Zhang YW, Jiang YS, Wang HY, Zhang YZ. Determination and analysis of sweet potato respiratory rate during storage period. J Sichuan Univ (Nat Sci Ed), 2010, 47 (5): 1131-1136]
3 张彩霞, 谢高地, 李士美, 盖力强 中国甘薯乙醇的资源潜力及空间分布. 资源科学, 2010, 32 (3): 505-511 [Zhang CX, XIE GD, Li SM, Gai LQ. Resource potential of sweet potato ethanol and its spatial distribution in China. Resour Sci, 2010, 32: 505-511]
4 李鹏霞, 王炜, 胡花丽, 王毓宁, 刘春泉. 低温贮藏下两种甘薯的品质变化研究. 江西农业学报, 2009, 21 (4): 73-75 [Li PX, Wang W, Hu HL, Wang YN, Liu CQ. Changes of quality of two varieties of sweet potato stored at low temperature. Acta Agric Jiangxi, 2009, 21 (4): 73-75]
5 谢逸萍, 李洪民, 王欣. 贮藏期甘薯块根淀粉酶活性变化趋势. 江苏农业学报, 2008, 24 (4): 406-409 [Xie YP, Li HM, Wang X. Changes of amylase activities in sweetpotato roots during storage. Jiangsu J Agric Sci, 2008, 24(4): 406~409
6 Ishiguro K, Yahara S, Yoshimoto M. Changes in polyphenolic content and radical-scavenging activity of sweet potato (Ipomoea batatas L.) during storage at optimal and low temperatures. J Agric Food Chem, 2007, 55: 10773-10778
7 Luan YS, Zhang J, Liu DM, Li WL. Molecular characterization of sweet potato leaf curl virus isolate from China (SPLCV-CN) and its phylogenetic relationship with other members of the Geminiviridae. Virus Genes, 2007, 35: 379-385
8 Liang Y, Srivastava S, Rahman MH, Strelkov SE, Kav NNV. Proteorne changes in leaves of Brassica napus L. as a result of Sclerotinia sclerotiorum challenge. J Agric Food Chem, 2008, 56: 1963-1976
9 Aghaei K, Ehsanpour AA, Komatsu S. Proteome analysis of potato under salt stress. J Proteome Res, 2008, 7: 4858-4868
10 O’Farrell PH. High resolution two-dimensional electrophoresis of proteins. J Biol Chem, 1975, 250: 4007-4021
11 Holloway PJ, Arundel PH. High-resolution two-dimensional electrophoresis of plant proteins. Anal Biochem, 1988, 172: 8-15
12 Muller EC, Schumann M, Rickers A, Bommert K, Wittmann-Liebold B, Otto A. Study of Burkitt lymphoma cell line proteins by high resolution two-dimensional gel electrophoresis and nanoelectrospray mass spectrometry. Electrophoresis, 1999, 20: 320-330
13 Hajduch M, Rakwal R, Agrawal GK, Yonekura M, Pretova A. High-resolution two-dimensional electrophoresis separation of proteins from metal-stressed rice (Oryza sativa L.) leaves: drastic reductions/fragmentation of ribulose-1,5-bisphosphate carboxylase/oxygenase and induction of stress-related proteins. Electrophoresis, 2001, 22: 2824-2831
14 杨艳华, 崔为同, 刘晓勇, 朱克明, 陈克平. 水稻叶片蛋白质组双向电泳实验条件的改进. 应用与环境生物学报, 2012, 18 (5): 819-823 [Yang YH, Cui WT, Liu XY, Zhu KM, Chen KP. Improvement of two-dimensional electrophoresis of improvement of two-dimensional electrophoresis of rice (Oryza sativa L.) proteomics. Chin J Appl Environ Biol, 2012, 18 (5): 819-823]
15 Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem, 1976, 72: 248-254
16 Herbert B. Advances in protein solubilisation for two-dimensional electrophoresis. Electrophoresis, 1999, 20: 660-663
17 Chen W, Ji JG, Xu XM, He SZ, Ru BG. Proteomic comparison between human young and old brains by two-dimensional gel electrophoresis and identification of proteins. Int J Dev Neurosci 2003, 21: 209-216
18 Neuhoff V SR, Eibl H. Clear background and highly sensitive protein staining with Coomassie blue dyes in polyacrylamide gels. Electrophoresis,1985, 6: 427-448
19 刘伟霞, 潘映红. 适用于小麦叶片蛋白质组分析的样品制备方法. 中国农业科学, 2007, 40 (10): 2169-2176 [Liu WA, Pan YH. Sample preparation methods suitable for wheat leaf proteome analysis. Sci Agric Sin, 2007, 40 (10): 2169-2176]
20 Canovas FM, Dumas-Gaudot E, Recorbet G, Jorrin J, Mock HP, Rossignol M. Plant proteome analysis. Proteomics, 2004, 4: 285-298
21 Hari V. A method for the two-dimensional electrophoresis of leaf proteins. Anal Biochem, 1981, 113: 332-335
22 罗泽宇, 杨粤军, 刘选明. 拟南芥蛋白质组研究中双向电泳技术条件的优化. 激光生物学报, 2008, 17选择(4): 539-545 [Luo ZY, Yang YJ, Liu XM. Optimization on two-dimensional electrophoresis for proteomic study in Arabidopsis thaliana. Acta Laser Biol Sin, 2008, 17(4): 539-545
23 Gilmore JM, Washburn MP. Advances in shotgun proteomics and the analysis of membrane proteomes. J Proteomics, 2010, 73: 2078-2091
24 Gorg A, Postel W, Domscheit A, Gunther S. Two-dimensional electrophoresis with immobilized pH gradients of leaf proteins from barley (Hordeum vulgare): method, reproducibility and genetic aspects. Electrophoresis, 1988, 9: 681-692
25 王小丽, Gonzalez perez P, 叶俊, 黄丹枫. 土壤宏蛋白质组学蛋白质提取方法及其应用. 应用与环境生物学报, 2012, 18 (4): 691-696 [Wang XL, Gonzalez perez Pablo, Ye J, Huang DF. Methods of protein extraction from soil and their potential application in study of soil metaproteome. Chin J Appl Environ Biol, 2012, 18 (4): 691-696]


Last Update: 2014-05-04