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Prokaryotic Expression, Purification, Crystalization and Electron Paramagnetic Resonance of a Mutant T179A of Alternative Oxidase

Abstract

交替氧化酶(AlternativeOxidase,AOX),广泛存在于高等植物中,是植物线粒体内膜上呼吸链中抗氰呼吸途径的末端氧化酶。AOX参与植物多种生理代谢调节,例如调节呼吸代谢、抑制活性氧的形成、抵抗逆境以及调节细胞凋亡等。然而,有关AOX的调控机理尚不清楚,对其结构也不完全清楚,目前仅有其二级结构模型的两种假说:SUM模型和AN模型,而AOX三级结构研究一直没有结果,这阻碍了对AOX结构与功能的研究。本研究利用DNA重组技术,克隆AOX基因及其第179位苏氨酸(T)突变为丙氨酸(A)的突变基因至原核表达载体pET-15b,运用原核表达系统BL21和FN102,经异丙基β-D-硫代半乳糖...Alternative Oxidase(AOX), widely existing in higher plants, is the terminal oxidase in the cyanide-resistant respiration pathway in plant mitochondria. AOX can regulate many physical metabolizable pathways in plant, for example regulation of respiration, restraining formation of reactive oxygen species, resistence of discipline and regulation of programmed cell death and so on. However, its regula...学位:理学博士院系专业:生命科学学院生物学系_植物学学号:2162006015328

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