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小定鞭藻新型锰超氧化物歧化酶(MnSOD)基因的分子克隆及其对氧化胁迫的响应。

Molecular cloning and oxidative-stress responses of a novel manganese superoxide dismutase (MnSOD) gene in the dinoflagellate Prorocentrum minimum.

机构信息

Department of Biotechnology, Sangmyung University, Seoul, 03016, South Korea.

Ocean Climate and Ecology Research Division, National Institute of Fisheries Science (NIFS), Busan, 46083, South Korea.

出版信息

Mol Biol Rep. 2019 Dec;46(6):5955-5966. doi: 10.1007/s11033-019-05029-6. Epub 2019 Aug 12.

Abstract

Dinoflagellate algae are microeukaryotes that have distinct genomes and gene regulation systems, making them an interesting model for studying protist evolution and genomics. In the present study, we discovered a novel manganese superoxide dismutase (PmMnSOD) gene from the marine dinoflagellate Prorocentrum minimum, examined its molecular characteristics, and evaluated its transcriptional responses to the oxidative stress-inducing contaminants, CuSO and NaOCl. Its cDNA was 1238 bp and contained a dinoflagellate spliced leader sequence, a 906 bp open reading frame (301 amino acids), and a poly (A) tail. The gene was coded on the nuclear genome with one 174 bp intron; signal peptide analysis showed that it might be localized to the mitochondria. Real-time PCR analysis revealed an increase in gene expression of MnSOD and SOD activity when P. minimum cells were separately exposed to CuSO and NaOCl. In addition, both contaminants considerably decreased chlorophyll autofluorescence, and increased intracellular reactive oxygen species. These results suggest that dinoflagellate MnSOD may be involved in protecting cells against oxidative damage.

摘要

甲藻是具有独特基因组和基因调控系统的微型真核生物,是研究原生动物进化和基因组学的有趣模型。本研究从海洋甲藻原甲藻中发现了一种新型锰超氧化物歧化酶(PmMnSOD)基因,对其分子特征进行了研究,并评估了其对氧化应激诱导污染物 CuSO 和 NaOCl 的转录响应。其 cDNA 为 1238bp,包含一个甲藻剪接先导序列、一个 906bp 的开放阅读框(301 个氨基酸)和一个 poly(A)尾。该基因位于核基因组上,有一个 174bp 的内含子;信号肽分析表明,它可能定位于线粒体。实时 PCR 分析表明,当 P. minimum 细胞分别暴露于 CuSO 和 NaOCl 时,MnSOD 的基因表达和 SOD 活性增加。此外,这两种污染物都显著降低了叶绿素自发荧光,并增加了细胞内的活性氧。这些结果表明,甲藻 MnSOD 可能参与了细胞对氧化损伤的保护。

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