Molecular cloning, mRNA expression and characterization of membrane-bound hemoglobin in oriental river prawn Macrobrachium nipponense

文献类型: 外文期刊

第一作者: Sun, Shengming

作者: Sun, Shengming;Fu, Hongtuo;Zhu, Jian;Ge, Xianping;Xuan, Fujun;Wu, Xugan

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关键词: Macro brachium nipponense;Membrane-bound hemoglobin;Hypoxia;Oxidative stress;Gill

期刊名称:COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY A-MOLECULAR & INTEGRATIVE PHYSIOLOGY ( 影响因子:2.32; 五年影响因子:2.465 )

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收录情况: SCI

摘要: Most hemoglobins are respiratory proteins and are ubiquitous in animals, bacteria, fungi, protists, and plants. In this study, we describe a membrane-bound hemoglobin in the oriental river prawn Macrobrachium nipponense (MnHb), which also expresses hemocyanin. MnHb cDNA was cloned using the rapid amplification of cDNA ends (RACE) approach, which afforded a 1201 bp gene encoding a 193 amino acid polypeptide. Bioinformatic evaluation suggested MnHb is membrane anchored by N-myristoylation, and immunofluorescence confirmed its location in the membrane of chief cells in the gill. The effect of hypoxia on MnHb expression was investigated, and reverse transcription PCR (RT-PCR) and Western blotting showed that MnHb was expressed almost exclusively in the gill. Quantitative RT-PCR revealed a significant increase in expression after 6 h of hypoxia, and levels peaked at 24 h due to oxidative stress. Exposure of cultured prawns to the stress inducer H2O2 significantly up regulated the expression of MnHb in a dose-dependent manner. MnHb may have a role in protecting cell membrane lipids from damage by reactive oxygen species. (C) 2017 Elsevier Inc. All rights reserved.

分类号: Q4`Q5`R33

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