苹果叶片蛋白质双向电泳样品制备方法的比较 |
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作者姓名: | 曾广娟 李春敏 张新忠 滕云龙 董文轩 |
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作者单位: | 沈阳农业大学园艺学院,沈阳 110161;河北省农林科学院昌黎果树研究所,昌黎 066600; 中国农业大学园艺植物研究所,北京 100193; |
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基金项目: | 河北省自然科学基金资助项目 |
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摘 要: | 为探索适用于苹果叶片蛋白质组学研究的样品制备方法,比较了三氯乙酸/丙酮沉淀法(TCA)、酚-甲醇/醋酸铵沉淀法和Tris-HCl提取法等3种总蛋白质的提取方法。制备的样品经双向电泳分离采用银染显色。结果3种方法分别得到450、374和1032个蛋白质点。认为Tris-HCl提取法为最适方法,所得图谱背景清晰,蛋白质信息量最大。
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关 键 词: | 苹果 蛋白质组 双向电泳 |
Comparison of Protein Extraction Methods for Two-dimensional Electrophoresis in Apple Leaf |
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Authors: | Zeng Guangjuan Li Chunmin Zhang Xinzhong Teng Yunlong Dong Wenxuan |
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Institution: | College of Horticulture, Shenyang Agricultural Univercity, Shenyang 110161; Changli Institute of Pomology, Hebei Academy of Agricultural and Forestry Science, Changli 066600; Institute for Horticultural Plants, China Agricultural University, Beijing |
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Abstract: | In order to develop an efficient protein extraction method suitable for apple proteome analysis, three protocols for total protein extraction in apple leaf, trichloroacetic acid/acetone precipitation (TCA), phenol extraction methanol/ammonium acetate precipitation and Tris-HCl extraction were compared. The extracts were separated by two-dimensional electrophoresis (2-DE) followed by silver staining. The results showed that 450, 374 and 1032 protein spots were detected respectively on 2-DE gels. The Tris-HCl extraction was the most appropriate for apple leaf proteome analysis on account of the highest resolution and more informative 2-DE gels. |
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