引用本文:	赵宇鹏,周伟,赵佩佩,才金玲,朱大玲,潘光华,刘洪艳,王广策.适用于条斑紫菜的双向电泳样品制备方法[J].海洋科学,2013,37(5):19-24.

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适用于条斑紫菜的双向电泳样品制备方法
赵宇鹏¹, 周伟^2,3, 赵佩佩^2,3, 才金玲¹, 朱大玲¹, 潘光华¹, 刘洪艳¹, 王广策²
1.天津科技大学海洋科学与工程学院;2.中国科学院海洋研究所;3.中国科学院大学

摘要:

为了建立适于条斑紫菜(Porphyra yezoensis)蛋白质组学分析的的样品提取方法, 作者以条斑紫菜叶状体为材料, 比较了饱和硫酸铵沉淀法、三氯乙酸/丙酮沉淀法和酚提取方法3 种方法对条斑紫菜叶状体总蛋白的提取效果。结果表明, 针对条斑紫菜叶状体细胞中含有大量多糖, 多酚化合物, 色素等干扰物质的特点, 通过酚提取方法能有效去除这些干扰杂质, 得到清晰、稳定、干净的双向电泳图谱和相对较多的蛋白质点数, 为提高其他大型海藻的总可溶性蛋白提取效率提供了重要参考。

关键词: 条斑紫菜(Porphyra yezoensis) 双向电泳蛋白提取硫酸铵三氯乙酸酚

DOI：

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基金项目:海洋公益性项目(201105023-7); 江苏省前瞻性联合研究项目(BY2011188); 中国科学院院先导项目(XDA05030401)海洋公益子项目(201105008-2)

A protein extraction method suitable for two-dimensional electrophoresis analysis of Porphyra yezoensis

Abstract:

Three different methods including saturated ammonium sulfate precipitation, trichloroacetic acid/acetone precipitation (TCA) and phenol extraction (Phe) have been compared to determine their efficiency in extracting total proteins from thalli of Porphyra yezoensis by two-dimensional electrophoresis (2-DE) analysis. The results showed that phenol protein extraction method gave the distinct stable and clear 2-DE map and was capable of generating more protein spots under the situation that there are many interfering substances of salinity, polysaccharide and pigments in the cells of P. yezoensis thalli. Findings from this study are of great significance for developing effective protein extraction methods for other macroalgae.

Key words: Porphyra yezoensis two-dimensional electrophoresis protein extraction ammonium sulfate trichloroacetic acid phenol