| 烟草根系蛋白质双向电泳样品制备方法的比较 |
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| 引用本文： | 林世锋,张拓,史跃伟,王东茂,王志红,杨志晓,谢升东,魏开华,任学良.烟草根系蛋白质双向电泳样品制备方法的比较[J].贵州农业科学,2012(8):71-74. |
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| 作者姓名： | 林世锋 张拓 史跃伟 王东茂 王志红 杨志晓 谢升东 魏开华 任学良 |
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| 作者单位： | 贵州省烟草科学研究所;北京蛋白质组研究中心 蛋白质组学国家重点实验室 |
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| 基金项目： | 中国烟草总公司重点项目“烤烟育种新技术与新一代烤烟新品种选育研究”(2010-221);贵州省科技厅农业攻关项目“烤烟种质资源鉴定与创新及新品种选育研究”(2011-3047) |
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| 摘 要： | 为探索适于烟草根系蛋白质组学研究的样品制备方法,以栽培品种K326生长期根系为材料,比较了常规裂解液法、TCA/丙酮沉淀法、Trizol抽提法和酚法等4种总蛋白质的提取方法。对制备的样品进行聚丙烯酰胺凝胶电泳(SDS-PAGE)及双向电泳(2-DE)检测。结果表明:酚法最佳,所获蛋白在单向SDS-PAGE中形成条带数目多而清晰;经双向电泳分离用银染显色,可检测约548个蛋白点,图谱分辨率较好,蛋白点清晰,主要集中在等电点pH5～8、相对分子量25.0～70.0kD,可有效解析烟草根系的蛋白质组。
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| 关 键 词： | 烟草 根系 蛋白质 蛋白质组 蛋白提取 双向电泳 制备方法 |
Comparison of Protein Extraction Methods for Two-dimensional Electrophoresis in Tobacco Root |
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| LIN Shi-feng,ZHANG Tuo,SHI Yue-wei,WANG Dong-mao,WANG Zhi-hong,YANG Zhi-xiao,XIE Sheng-dong,WEI Kai-hua,RENXue-liang.Comparison of Protein Extraction Methods for Two-dimensional Electrophoresis in Tobacco Root[J].Guizhou Agricultural Sciences,2012(8):71-74. |
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| Authors: | LIN Shi-feng ZHANG Tuo SHI Yue-wei WANG Dong-mao WANG Zhi-hong YANG Zhi-xiao XIE Sheng-dong WEI Kai-hua RENXue-liang |
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| Institution: | 1 (1.Guizhou Tobacco Research Institute,Guiyang,Guizhou 550081;2.Beijing Proteomics Research Center,State Key Laboratory of Proteomics,Beijing 102206,China) |
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| Abstract: | In order to develop an efficient protein extraction method suitable for tobacco proteome analysis,four protocols for total protein extraction in tobacco root,lysis-buffer method,TCA/acetone precipitation method,Trizol extraction method and phenol method were assessed with tobacco cultivar K326 root at growth stage as experiment materials.The protein samples were separated by SDS-PAGE and two-dimensional electrophoresis(2-DE).It was found that the phenol method was the best one resulting in the most protein bands and the best resolution in SDS-PAGE.About 548 clear protein spots were detected by 2-DE and silver staining with isoelectric points at pH 5～8,and relative molecular weight of protein ranged from 25.0 kD to 70.0 kD. |
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| Keywords: | tobacco root proteome protein extraction two-dimensional electrophoresis |
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