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首页> 美国卫生研究院文献>Plant Physiology >Special Issue on Reactive Oxygen Species: Expression and Characterization of a Redox-Sensing Green Fluorescent Protein (Reduction-Oxidation-Sensitive Green Fluorescent Protein) in Arabidopsis
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Special Issue on Reactive Oxygen Species: Expression and Characterization of a Redox-Sensing Green Fluorescent Protein (Reduction-Oxidation-Sensitive Green Fluorescent Protein) in Arabidopsis

机译：活性氧物种的特殊问题：拟南芥中氧化还原敏感的绿色荧光蛋白（还原-氧化敏感的绿色荧光蛋白）的表达和表征

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摘要

Arabidopsis (Arabidopsis thaliana) was transformed with a redox-sensing green fluorescent protein (reduction-oxidation-sensitive green fluorescent protein [roGFP]), with expression targeted to either the cytoplasm or to the mitochondria. Both the mitochondrial and cytosolic forms are oxidation-reduction sensitive, as indicated by a change in the ratio of 510 nm light (green light) emitted following alternating illumination with 410 and 474 nm light. The 410/474 fluorescence ratio is related to the redox potential (in millivolts) of the organelle, cell, or tissue. Both forms of roGFP can be reduced with dithiothreitol and oxidized with hydrogen peroxide. The average resting redox potentials for roots are −318 mV for the cytoplasm and −362 mV for the mitochondria. The elongation zone of the Arabidopsis root has a more oxidized redox status than either the root cap or meristem. Mitochondria are much better than the cytoplasm, as a whole, at buffering changes in redox. The data show that roGFP is redox sensitive in plant cells and that this sensor makes it possible to monitor, in real time, dynamic changes in redox in vivo.
机译：用氧化还原敏感的绿色荧光蛋白（还原氧化敏感的绿色荧光蛋白[roGFP]）转化拟南芥（Arabidopsis thaliana），其表达靶向细胞质或线粒体。线粒体形式和胞质形式均对氧化还原敏感，如通过在410和474 nm的光交替照射后发出的510 nm的光（绿光）的比率变化所表明的。 410/474荧光比与细胞器，细胞或组织的氧化还原电位（以毫伏为单位）有关。两种形式的roGFP都可以用二硫苏糖醇还原，并用过氧化氢氧化。根的平均静息氧化还原电势对于细胞质为-318 mV，对于线粒体为-362 mV。拟南芥根的延伸区具有比根冠或分生组织更氧化的氧化还原状态。整体而言，线粒体在氧化还原的缓冲方面比细胞质要好得多。数据表明，roGFP在植物细胞中对氧化还原敏感，并且该传感器使实时监测体内氧化还原的动态变化成为可能。

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