两个谷胱甘肽过氧化物酶基因在水稻中的功能分化

Jingxin Wei, Hai-Ling Yang
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摘要

植物谷胱甘肽过氧化物酶(GPX)是一组调节细胞和组织中活性氧水平并保护细胞和组织免受氧化损伤的酶。本研究从水稻中克隆了两个GPX基因OsGPX3和OsGPX4。OsGPX3和OsGPX4基因分别编码238和234个氨基酸残基的蛋白,计算分子质量分别为25.84 kDa和25.07 kDa。基因组序列分析显示,OsGPX3和OsGPX4含有5个内含子。反转录PCR结果显示,OsGPX3和OsGPX4是satisatia的组成基因,在大肠杆菌中过表达OsGPX3和OsGPX4蛋白,并通过ni亲和层析纯化。OsGPX3和OsGPX4蛋白对底物H2O2、tBOOH和COOH具有酶促活性。然而,OsGPX3对这三种底物的酶活性高于OsGPX4,这表明OsGPX3和OsGPX4基因在功能上存在差异。
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Functional Divergence of Two Glutathione Peroxidase Genes in Oryza sativa
Plant glutathione peroxidases(GPX) are a group of enzymes that regulate levels of reactive oxygen species in cells and tissues,and protect them against oxidative damage.In this study,two GPX genes(OsGPX3 and OsGPX4) were cloned from Oryza sativa.The OsGPX3 and OsGPX4 genes encoded two proteins of 238 and 234 amino acid residues,with calculated molecular masses of 25.84 kDa and 25.07 kDa,respectively.Genomic sequence analysis showed OsGPX3 and OsGPX4 contained five introns.Reverse transcription PCR revealed that OsGPX3 and OsGPX4 were constitutive expression genes in O.sativa.The OsGPX3 and OsGPX4 proteins were overexpressed in E.coli,and were purified by Ni-affinity chromatography.The OsGPX3 and OsGPX4 proteins showed enzymatic activities towards substrates H2O2,tBOOH and COOH.However,OsGPX3 showed higher enzymatic activities to the three substrates than did OsGPX4,suggesting functional divergence between the OsGPX3 and OsGPX4 genes.
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