An Extraction Method Suitable for Two-Dimensional Electrophoresis of Low-abundant Proteins from Ginseng Roots
Four lysis buffers, which contain different concentration ratio of urea and thiourea, and three extraction methods (sonication extraction, osmotic extraction, and freeze–thaw extraction) were used to optimize the extraction conditions of low-abundant prot
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An Extraction Method Suitable for TwoDimensional Electrophoresis of Lowabundant Proteins from Ginseng Roots Rui Jiang, Liwei Sun, Rui Ma, Xiujuan Lei and Daqing Zhao
Abstract Four lysis buffers, which contain different concentration ratio of urea and thiourea, and three extraction methods (sonication extraction, osmotic extraction, and freeze–thaw extraction) were used to optimize the extraction conditions of low-abundant proteins from ginseng roots based on protein extraction efficiency, the good quality of the 2-DE patterns, and the results of mass spectrometry (MS) identification. Our results have shown that when lysis buffer II (7 M urea, 2 M thiourea, 2 % CHAPS, 1 % PMSF, and 1 % protease inhibitor) combined with sonication was used, higher protein extraction efficiency, lower background, more protein spots (987 ± 31), and higher spot intensities of lowabundant proteins were obtained, which indicated that the optimized protocol was suitable for proteomic studies of ginseng root. Moreover, those results provided the foundation for further researches on the significant functional proteins of ginseng roots. Keywords Two-dimensional electrophoresis proteins Protein extraction
Ginseng root
Low-abundant
R. Jiang R. Ma D. Zhao (&) Changchun University of Chinese Medicine, Changchun 130117, People’s Republic of China e-mail: [email protected] R. Jiang L. Sun (&) Beihua University, Jilin 132013, People’s Republic of China e-mail: [email protected] X. Lei The Chinese Academy of Agricultural Sciences Institute of Specialty, Jilin 132011, People’s Republic of China
T.-C. Zhang et al. (eds.), Proceedings of the 2012 International Conference on Applied Biotechnology (ICAB 2012), Lecture Notes in Electrical Engineering 251, DOI: 10.1007/978-3-642-37925-3_150, Ó Springer-Verlag Berlin Heidelberg 2014
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150.1 Introduction Panax ginseng C. A. Meyer is a well-known traditional Chinese medicine (TCM) and has been used in China, Korea, and other Asian countries for over 2,000 years. Although ginsenosides are considered to be the main active pharmacological compounds in ginseng roots [1], various pharmacological effects of ginseng proteins have been reported now, such as antioxidative activity [2], serum lipid and blood pressure reducing activity [3, 4], antifatigue activity [5], radioresistance effect [6], regulating immune function [7], antifungal effect [8]. However, less than 100 functional proteins of ginseng roots have been reported [9–13]. It would be quite necessary to systematically study the ginseng proteins that performed important bioactive functional effects. With the development of functional genomics and proteomics technology, the plant proteomics technology has been improved rapidly. It has been used to rapidly identify hundreds of proteins, discover special functional proteins, elucidate the underlying mechanisms of plant growth metabolism, regulation, and stress resistance. Plant proteomics is also becoming an essential technology in the study of TCM functi
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