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首页> 外文期刊>Molecular and Biochemical Parasitology >Molecular cloning and characterization of a phospholipid hydroperoxide glutathione peroxidase gene from a blood fluke Schistosoma japonicum
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Molecular cloning and characterization of a phospholipid hydroperoxide glutathione peroxidase gene from a blood fluke Schistosoma japonicum

机译：血液氟氏血吸虫磷酸盐磷酸酸缩水素基因的分子克隆与表征

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摘要

Phospholipid hydroperoxide glutathione peroxidase (PHGPx) is a major antioxidant enzyme and plays critical roles in the protection of cells against oxidative stress by catalysing reduction of lipid hydroperoxides. A full-length cDNA sequence corresponding to GPx gene from Schistosoma japonicum (designated SjGPx) was isolated and characterized. SjGPx contained an in-frame TGA codon for selenocysteine (Sec) and a concurrent Sec insertion sequence in its 3'-untranslated region. Protein encoded by SjGPx demonstrated a primary structure characteristic to the PHGPx family, including preservation of catalytic domains and absence of the subunit interaction domains. Phylogenetic analysis revealed that the SjGPx was highly related to the other PHGPx-related members, and clustered into the trematode subclade II. Semi-quantitative reverse transcription PCR and western blotting showed that the SjGPx was mainly expressed in the female adults and eggs. RNA interference was employed to investigate the effects of knockdown of SjGPx. SjGPx expression level was significantly reduced on the 5th day post-RNAi. We observed a 53.86% reduction in total GPx activity and the eggs severely deformed. Oxidative stimulation of viable worms with H2O2 or paraquat resulted in 1.6- to 2.1-fold induction of the GPx activity. Our results revealed that the SjGPx protein is selenium-dependent PHGPx, which might actively participate in the detoxification of oxidative damage during egg production. (C) 2015 Elsevier B.V. All rights reserved.
机译：磷脂氢过氧化物谷胱甘肽过氧化物酶（PHGPX）是主要的抗氧化酶，并通过催化脂质过氧化钠的降低来保护细胞免受氧化应激的关键作用。分离出与血吸虫血吸虫（指定SJGPX）的GPX基因对应的全长cDNA序列，并表征。 SJGPX含有用于Selenocysteine（SEC）的框架TGA密码子和其3' - 未转换区域中的并发秒插入序列。由SJGPX编码的蛋白质证明了PHGPX家族的主要结构特征，包括保存催化结构域并没有亚基相互作用域。系统发育分析表明，SJGPX与其他与其他PHGPX相关成员高度相关，并聚集到Threadode亚克片II中。半定量逆转录PCR和Western印迹表明，SJGPX主要在女性成人和鸡蛋中表达。使用RNA干扰来研究SJGPX敲低的影响。在RNAi后第5天显着降低了SJGPX表达水平。我们观察到总GPX活性减少53.86％，鸡蛋严重变形。具有H 2 O 2或百草枯的可行蠕虫的氧化刺激导致GPX活性的1.6至2.1倍。我们的研究结果表明，SJGPX蛋白是依赖于硒的PHGPX，这可能会积极参与鸡蛋生产过程中氧化损伤的解毒。 （c）2015 Elsevier B.v.保留所有权利。

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