Characterization and expression of cytoplasmic copper/zinc superoxide dismutase (CuZn SOD) gene under temperature and hydrogen peroxide (H2O2) in rotifer Brachionus calyciflorus |
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Authors: | Jianghua Yang Siming DongQichen Jiang Qin SiXuezhou Liu Jiaxin Yang |
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Affiliation: | Jiangsu Province Key Laboratory for Biodiversity & Biotechnology and Jiangsu Province Key Laboratory for Aquatic Live Food, School of Biological Sciences, Nanjing Normal University, Nanjing, Jiangsu 210046, PR China |
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Abstract: | Superoxide dismutase (SOD, EC 1.15.1.1) is an important antioxidant enzyme that protects organs from damage by reactive oxygen species (ROS). We cloned cDNA encoding SOD activated with copper/zinc (CuZn SOD) from the rotifer Brachionus calyciflorus Pallas. The full-length cDNA of CuZn SOD was 692 bp and had a 465 bp open reading frame encoding 154 amino acids. The deduced amino acid sequence of B. calyciflorus CuZn SOD showed 63.87%, 60.00%, 59.74% and 48.89% similarity with the CuZn SOD of the Ctenopharyn godonidella, Schistosoma japonicum, Drosophila melanogaster and Caenorhabditis elegans, respectively. The phylogenetic tree constructed based on the amino acid sequences of CuZn SODs from B. calyciflorus and other organisms revealed that rotifer is closely related to nematode. Analysis of the expression of CuZn SOD under different temperatures (15, 30 and 37 °C) revealed that its expression was enhanced 4.2-fold (p < 0.001) at 30 °C after 2 h, however, the lower temperature (15 °C) promoted CuZn SOD transiently (4.1-fold, p < 0.001) and then the expression of CuZn SOD decreased to normal level (p > 0.05). When exposed to H2O2 (0.1 mM), CuZn SOD, manganese superoxide dismutase (Mn SOD) and catalase (CAT) gene were upregulated, and in addition, the mRNA expression of CuZn SOD gene was induced instantaneously after exposure to vitamin E. It indicates that the CuZn SOD gene would be an important gene in response to oxidative and temperature stress. |
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Keywords: | β-actin, beta-actin BBM, Bolds Basal Medium CR, calorie restriction CAT, catalase Ct, cycle threshold CuZn SOD, copper/zinc SOD EPA, artificial freshwater medium GPx, glutathione peroxidase GR, glutathione reductase GSH, glutathione GST, glutathione S-transferase H2O2, hydrogen peroxide Mn SOD, manganese SOD O2-, superoxide anion ORF, open reading frame RNAi, RNA interfere ROS, reactive oxygen species RT-PCR, Real-time PCR SOD, superoxide dismutase |
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