| Protein Extraction Methods for Two-Dimensional Electrophoresis from Baphicacanthus cusia （Nees）Bremek Leaves-A Medicinal Plant with High Contents of Interfering Compounds |
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| 引用本文： | XIANG Xiao-liang,NING Shu-ju,JIANG Xia,GONG Xiao-gui,ZHU Ren-lei,WEI Dao-zhi. Protein Extraction Methods for Two-Dimensional Electrophoresis from Baphicacanthus cusia （Nees）Bremek Leaves-A Medicinal Plant with High Contents of Interfering Compounds[J]. 中国农业科学(英文版), 2010, 9(10): 1530-1537. DOI: 10.1016/S1671-2927(09)60247-9 |
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| 作者姓名： | XIANG Xiao-liang NING Shu-ju JIANG Xia GONG Xiao-gui ZHU Ren-lei WEI Dao-zhi |
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| 作者单位： | XIANG Xiao-liang,GONG Xiao-gui,ZHU Ren-lei,WEI Dao-zhi(School of Life Sciences,Fujian Agriculture and Forestry University,Fuzhou 350002.P.R.China);NING Shu-ju(School of Crop Sciences,Fujian Agriculture and Forestry University,Fuzhou 350002,P.R.China);JIANG Xia(College of Chemistry and Materials Science,Fujian Normal University.Fuzhou 350002,P.R.China) |
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| 基金项目： | Acknowledgements This work was supported by the Department of Education （JA05238） and the Key Project on Ecology of Fujian Province, China （0608537）. We are grateful to the members of the laboratory at Agroecological Institute, Fujian Agriculture and Forestry University for technical support and to Ms. Xiong Jun for her valuable assistance. |
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| 摘 要： | Protein extraction is a critical step for two-dimensional electrophoresis （2-DE）. Different plant samples require different and adaptive protein extraction protocols. The leaves of medicinal plant, Baphicacanthus cusia （Nees） Bremek are notoriously recalcitrant to common protein extraction methods due to high levels of interfering compounds （especially the secondary metabolites and pigments）. This study was aimed to establish a routine procedure for the proteomic analysis ofB. cusia leaves, and a new protocol for the protein extraction was developed by optimizing trichloroacetic acid （TCA）/ acetone extraction method. The efficiency of this protocol was demonstrated by comparison with 3 published protein extraction methods （chloroform/acetone, Mg/NP-40, Tris-base/acetone）. The results showed that the optimized TCA/ acetone precipitation extraction method gave a relatively high protein yield （9.263 mg g^-1 fresh weight）, high-resolution separation, clear protein profiles, the highest proteins spots （1 31 t protein spots）, and displayed less contamination in 2- DE gels. Therefore, the results suggested that the optimized TCA/acetone method was the most effective among the 4 methods for B. cusia leaves.
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| 关 键 词： | 蛋白质提取 提取方法 二维电泳 药用植物 化合物 干扰 次生代谢产物 自主知识产权 |
| 收稿时间： | 2010-01-21 |
Protein Extraction Methods for Two-Dimensional Electrophoresis from Baphicacanthus cusia (Nees) Bremek Leaves-A Medicinal Plant with High Contents of Interfering Compounds |
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| XIANG Xiao-liang,NING Shu-ju,JIANG Xia,GONG Xiao-gui,ZHU Ren-lei,WEI Dao-zhi. Protein Extraction Methods for Two-Dimensional Electrophoresis from Baphicacanthus cusia (Nees) Bremek Leaves-A Medicinal Plant with High Contents of Interfering Compounds[J]. 《Agricultural Sciences in China》, 2010, 9(10): 1530-1537. DOI: 10.1016/S1671-2927(09)60247-9 |
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| Authors: | XIANG Xiao-liang NING Shu-ju JIANG Xia GONG Xiao-gui ZHU Ren-lei WEI Dao-zhi |
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| Affiliation: | 1. School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, P.R. China;2. School of Crop Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, P.R. China;3. College of Chemistry and Materials Science, Fujian Normal University, Fuzhou 350002, P.R. China;1. The Interdisciplinary Department of Social Sciences, Bar-Ilan University, Ramat Gan, Israel;2. Psychology Department, Ariel University, Ariel; Academic College, Gush Etzion, Israel |
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| Abstract: | Protein extraction is a critical step for two-dimensional electrophoresis (2-DE). Different plant samples require different and adaptive protein extraction protocols. The leaves of medicinal plant, Baphicacanthus cusia (Nees) Bremek are notoriously recalcitrant to common protein extraction methods due to high levels of interfering compounds (especially the secondary metabolites and pigments). This study was aimed to establish a routine procedure for the proteomic analysis of B. cusia leaves, and a new protocol for the protein extraction was developed by optimizing tfichloroacetic acid (TCA)/acetone extraction method. The efficiency of this protocol was demonstrated by comparison with 3 published protein extraction methods (chloroform/acetone, Mg/NP-40, Tris-base/acetone). The results showed that the optimized TCA/acetone precipitation extraction method gave a relatively high protein yield (9.263 mg g<'-1> fresh weight), high-resolution separation, clear protein profiles, the highest proteins spots (1 311 protein spots), and displayed less contamination in 2DE gels. Therefore, the results suggested that the optimized TCA/acetone method was the most effective among the 4 methods for B. cusia leaves. |
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| Keywords: | Baphicacanthus cusia (Nees) Bremek medicinal plant protein extraction two-dimensional electrophoresis |
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