| 分蘖洋葱叶片总蛋白提取与双向电泳条件优化 |
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| 引用本文： | 刘淑芹,吴凤芝.分蘖洋葱叶片总蛋白提取与双向电泳条件优化[J].东北农业大学学报,2013(1):71-76. |
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| 作者姓名： | 刘淑芹 吴凤芝 |
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| 作者单位： | 东北农业大学园艺学院 |
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| 基金项目： | 国家自然科学基金(31172002);国家“973”专项(2009CB119004-05);大宗蔬菜产业技术体系专项(CARS-25-08) |
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| 摘 要： | 以分蘖洋葱叶片为试验材料,比较3种不同蛋白质提取方法对2-DE蛋白质双向电泳结果的影响,并对其中的蛋白上样量、SDS-PAGE凝胶浓度及双向电泳重复性进行筛选与优化。结果表明,与酚提取法和改良的酚提取法相比,TCA-丙酮提取法提取分蘖洋葱叶片总蛋白的操作简便,所得双向电泳图谱背景清晰,蛋白质点圆滑,数量及质量均较好,是一种切实可行的提取分蘖洋葱叶片总蛋白方法。应用Image Master 2D Platinum 7.0软件对图谱进行分析,凝胶上检测到超过650多个蛋白质点,不同凝胶之间蛋白质的匹配率达92.5%,具有较高的分辨率和重复性,建立一套适用于分蘖洋葱叶片总蛋白质组分析的双向电泳(2-DE)方法。
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| 关 键 词： | 分蘖洋葱 叶片蛋白 双向凝胶电泳 |
Improvement of extraction method and two-dimensional electrophoresis conditions for proteome study on potato onion leaf |
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| LIU Shuqin,WU Fengzhi.Improvement of extraction method and two-dimensional electrophoresis conditions for proteome study on potato onion leaf[J].Journal of Northeast Agricultural University,2013(1):71-76. |
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| Authors: | LIU Shuqin WU Fengzhi |
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| Institution: | (School of Horticulture,Northeast Agricultural University,Harbin 150030,China) |
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| Abstract: | In this study,common potato onion leaf was taken as experimental material,leaf protein sample of young potato onion prepared by three methods.Two-dimensional electrophoresis was optimized in regard to protein extraction,the suitable concentration of protein per IPG gel,repetitions of experiment and SDS-PAGE parameters.The results showed that the protein yield extracted by TCA-acetone extraction method was much higher than that by phenolic extraction and modified phenolic method.The SDS-PAGE electrophoresis demonstrated that the protein isolated by TCA-acetone extraction method exhibited clearer bands and higher resolution than that by the other method with a fuzzy,low resolution.More than 650 proteins were detected in each gel and 92.5% of them were matched with each other in different gels stained by Coomassie Brilliant Blue analyzed by Image Master 2D Platinum 7.0 software,indicating the protocol with high repeatability and resolution.2-DE(two-dimensional electrophoresis) protocol suited for the separation of proteins from potato onion leaf. |
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| Keywords: | potato onion leaf protein two-dimensional gel electrophoresis |
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