| 圆果种黄麻功能叶总蛋白提取方法及双向电泳体系的优化 |
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| 引用本文： | 陈富成,祁建民,徐建堂,陈涛,陶爱芬,林培清,陈美霞,郭英,李华丽.圆果种黄麻功能叶总蛋白提取方法及双向电泳体系的优化[J].作物学报,2011,37(2):369-373. |
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| 作者姓名： | 陈富成 祁建民 徐建堂 陈涛 陶爱芬 林培清 陈美霞 郭英 李华丽 |
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| 作者单位： | 福建农林大学/ 作物遗传育种与综合利用教育部重点实验室，福建福州350002 |
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| 基金项目： | 国家麻类产业技术体系建设，国家农业部公益性产业计划，农业部引进先进技术(948计划)项目 |
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| 摘 要： | 蛋白质提取方法和电泳条件是蛋白质组学研究中双向电泳的关键。本文初步建立起一套适合圆果种黄麻功能叶总蛋白双向电泳(2-DE)分析的高效提取方法及电泳条件，在双向电泳技术中，比较了TCA丙酮、Tris-base/丙酮、Tris-HCl的蛋白提取方法，并优化了上样量和样品制备方法。结果表明, TCA丙酮法提取蛋白质产量高，达79.0 mg g–1，比Tris-base/丙酮法和Tris-HCl的产量分别高出44.2%和114.1%。该法得到的2-DE图谱背景清晰，蛋白点数最多，约602个，较其他两种方法更适合用于黄麻功能叶总蛋白双向电泳分析；350 mg为最佳上样量。这为黄麻蛋白质组学后续研究奠定了基础。
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| 关 键 词： | 黄麻功能叶 提取方法 蛋白质双向电泳(2-DE) 优化上样量 |
| 收稿时间： | 2010-07-06 |
Optimization of Extraction Method and Two-Dimen-sional Electrophopresis Conditions for Proteome Analysis of Jute Functional Leaf |
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| CHEN Fu-Cheng,QI Jian-Min,XU Jian-Tang,CHEN Tao,TAO Ai-Fen,LIN Pei-Qing,CHEN Mei-Xia,GUO Ying,LI Hua-Li.Optimization of Extraction Method and Two-Dimen-sional Electrophopresis Conditions for Proteome Analysis of Jute Functional Leaf[J].Acta Agronomica Sinica,2011,37(2):369-373. |
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| Authors: | CHEN Fu-Cheng QI Jian-Min XU Jian-Tang CHEN Tao TAO Ai-Fen LIN Pei-Qing CHEN Mei-Xia GUO Ying LI Hua-Li |
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| Institution: | Key Laboratory of Ministry of Education for Genetics, Breeding and Multiple Utilization of Crops, Fujian Agriculture and Forestry University, Fuzhou 350002, China |
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| Abstract: | The protein extraction methods and electrophoresis conditions are the key facts of two-dimensional electrophoresis in proteomics.In this study,in order to set up a suitable method for jute leaf total protein extraction and the optimized condition for two-dimensional electrophoresis,we compared the protein extraction methods using TCA acetone,Tris-base acetone and Tris-HCl.We also optimized the sample loading amount and sample preparation approach.The results showed that protein productivity reached 79.0 mg ... |
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| Keywords: | Jute leaf Extraction method Protein two-dimensional electrophoresis(2-DE) Optimizing of loading amount |
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