Establishment and optimization of two-dimensional electrophoresis (2-DE) technology for proteomic analysis of ramie
2013
Deng, G. (Huazhong Agricultural Univ., Hubei (China). Ministry of Agriculture) | Liu, L. (Huazhong Agricultural Univ., Hubei (China). Ministry of Agriculture) | Wang, H. (Huazhong Agricultural Univ., Hubei (China). Ministry of Education) | Lao, C. | Wang, B. | Zhu, C. | Peng, D.
This research aimed at establishment of a two-dimensional electrophoresis that is suitable for the proteomic analysis of ramie's various parts by optimizing the traditional TCA/acetone protein extraction method: protein purification (4 times volume of acetone), and IPG strips (pH 4-7, 17 cm). This technology is applied to the roots, stems and leaves of ramie. Compared with traditional method, this technology can effectively remove the non-protein impurities from ramie, the protein spots obtained from it (823 plus minus 15) significantly higher than the traditional (320 plus minus 8) method, and it is suitable for the different parts of ramie and the requirement of protein identification by Mass Spectrum; The number of protein in the stem, roots and leaves were 280 plus minus 6, 1093 plus minus 20 and 765 plus minus 13, respectively. The method used provides reliable technical support to the proteomic study of ramie that will aid the understanding of higher yield, higher quality, stress tolerance, and disease resistance.
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