首页 > 过刊浏览>2011年第19卷第6期 >519-523. DOI:10.3969/j.issn.1005-3395.2011.06.005
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红树植物秋茄叶片双向电泳技术体系的建立及优化
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国家海洋公益性行业科研专项经费项目(200805072; 201005012);浙江省近岸水域生物资源开发与保护重点实验室开放课题基金项目(2010F30003); 浙江省科技厅创新团队建设与人才培养项目(2009F20009); 浙江省海洋与渔业项目"高纬度红树林生态恢复关键技术应用示范"资助


Establishment and Optimization of 2 - DE Technique System in Leaf Proteome of Kandelia candel
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    摘要:

    对适用于秋茄(Kandelia candel)叶片蛋白质组学研究的双向电泳技术体系进行了优化。结果表明,采用酚抽提法提取的蛋白质浓度较低,约1.7 μg μL-1,SDS-PAGE电泳后几乎无条带;而采用改良TCA-丙酮沉淀法可显著提高提取液中蛋白质的含量,可达12.4 μg μL-1。秋茄叶片蛋白质主要分布在pH 4~7的范围内,最适于采用pH 3~10 NL的胶条,17 cm胶条的最适上样量为400 μg,经银染后可得蛋白点约(564±34)个。用TCA-丙酮沉淀法提取秋茄叶片全蛋白质组,按优化的参数进行双向电泳分离,获得了高分辨率、高重复性的图谱,可用于秋茄蛋白质组学研究。

    Abstract:

    In order to study the proteome of Kandelia candel leaves, a two-dimensional electrophoresis (2-DE) system was established and optimized. Compared with the low protein content (1.7 μg μL-1) extracted by phenol (Phe), the improved trichloroacetic acid/acetone precipitation (TCA) method could obviously increase the extracting efficiency up to 12.4 μg μL-1. The protein spots of Kandelia candel leaves mainly distributed in the range of pH 4-7. The optimal loading quantity of samples were 400 μg gel-1 for a 17 cm IPG strip (pH 3-10 NL), which can detect 564±34 protein spots after silver staining. Therefore, the high resolution and repeatable 2-DE map could be obtained by using TCA extract method and the optimum system, which could widely use in proteome study of Kandelia candel.

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于冯,郑春芳,施孟如,刘伟成,仇建标,冀德伟,黄丽,陈少波.红树植物秋茄叶片双向电泳技术体系的建立及优化[J].热带亚热带植物学报,2011,19(6):519~523

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  • 收稿日期:2011-03-07
  • 最后修改日期:2011-08-17
  • 录用日期:2011-09-05
  • 在线发布日期: 2011-11-18
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