| REN Hai,LI Jian,LI Jitao,YING Yu,GE Hongxing,LI Dongli,YU Tianji. 2015. Cloning of catalase and expression patterns of catalase and selenium-dependent glutathione peroxidase from Exopalaemon carinicauda in response to low salinity stress. Acta Oceanologica Sinica, 34(8):52－61 |
| Cloning of catalase and expression patterns of catalase and selenium-dependent glutathione peroxidase from Exopalaemon carinicauda in response to low salinity stress |
| 低盐胁迫对脊尾白虾过氧化氢酶和硒谷胱甘肽过氧化物酶基因表达的影响 |
| Received:March 11, 2014 Revised:August 28, 2014 |
| DOI：10.1007/s13131-015-0640-9 |
| Key words:Exopalaemon carinicauda catalase (CAT) selenium-dependent glutathione peroxidase (Se-GPx) cloning expression |
| 中文关键词: 脊尾白虾 过氧化氢 硒谷胱甘肽过氧化物酶 克隆 表达 |
| 基金项目:The Modern Agro-industry Technology Research System under contract No. CARS-47; the National High-tech R & D Program (863 Program) of China under contract No. 2012AA10A409; the Special Fund for Independent Innovation of Shandong Province under contract No. 2013CX80202; the Special Fund for Agro-scientific Research in the Public Interest under contract No. 201103034. |
| Author Name | Affiliation | E-mail | | REN Hai | College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China
Key Laboratory of Sustainable Development of Marine Fisheries of Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China | | | LI Jian | Key Laboratory of Sustainable Development of Marine Fisheries of Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China | lijian@ysfri.ac.cn | | LI Jitao | Key Laboratory of Sustainable Development of Marine Fisheries of Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China | | | YING Yu | Key Laboratory of Sustainable Development of Marine Fisheries of Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China | | | GE Hongxing | Key Laboratory of Sustainable Development of Marine Fisheries of Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China | | | LI Dongli | Key Laboratory of Sustainable Development of Marine Fisheries of Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China | | | YU Tianji | Key Laboratory of Sustainable Development of Marine Fisheries of Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China | |
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| Abstract: |
| Catalase (CAT) and selenium-dependent glutathione peroxidase (Se-GPx) play a vital role in protecting organisms against various oxidative stresses by eliminating H2O2. The objective of this paper is to evaluate the roles of these antioxidant molecules in the ridgetail white prawn Exopalaemon carinicauda in response to low salinity stress. A complementary DNA (cDNA) containing the complete coding sequence of CAT was cloned from the hepatopancreas using reverse-transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends. The full-length cDNA of CAT (2 649 bp) contains a 5'-untranslated region (UTR) of 78 bp, a 3'- UTR of 1 017 bp, with a poly (A) tail, and an open reading frame of 1 554 bp encoding a 517-amino-acid polypeptide with predicted molecular mass of 58.46 kDa and estimated isoelectric point of 6.64. This CAT sequence contained the proximal active site signature (60FDRERIPERVVHAKGAG76), proximal heme-ligand signature sequence (350RLFSYPDTH358) and three catalytic amino acid residues (His71, Asn144 and Tyr354). Sequence comparison showed that the CAT deduced amino acid sequence of E. carinicauda shared 68%-92% of identities with those of other species. Quantitative real-time PCR analysis revealed that CAT mRNA was widely expressed in the hepatopancreas (highest), hemocyte, eyestalk, heart, gill, muscle, ovary and stomach. Under low salinity stress, CAT and GPx mRNA expression levels both in the gill and hepatopancreas increased significantly at the first 48 h and 6 h respectively, indicating a tissue- and time-dependent antioxidant response in E. carinicauda. All these results indicate that E. carinicauda CAT is a member of the CAT family and might be involved in the acute response against low salinity stress. |
| 中文摘要: |
| 过氧化氢酶(CAT)和硒谷胱甘肽过氧化物酶(Se-GPx)在抗氧化防御体系中扮演重要的角色,在清除多余的活性氧自由基(ROS)防止机体氧化损伤方面具有重要的作用.本文利用RACE-PCR方法首次从脊尾白虾肝胰腺组织中克隆获得了CAT cDNA全长序列,该基因全长2649 bp,包括5′非编码区(UTR) 78 bp、开放阅读框(ORF)1554 bp和3′非编码区(UTR) 1017 bp,共编码517个氨基酸,预测分子量为58.46 kDa,理论等电点为6.64,利用PROSITE tools预测CAT基因的功能位点,发现该氨基酸序列包括酶活性中心序列“FDRERIPERVVHAKGAG”,亚铁血红素结合信号序列“RLFSYPDTH”和 3个催化位点残基His71,Asn144和Tyr354.同源性分析表明,脊尾白虾CAT氨基酸序列与其他物种的相似性为68%-92%.荧光定量PCR分析结果表明,CAT基因在肝胰腺、血细胞、心脏、肌肉、卵巢、鳃、胃和眼柄均有表达,其中在肝胰腺中表达量最高.低盐胁迫后脊尾白虾鳃和肝胰腺中CAT和GPx基因分别在48 h和6 h达到最大值,且与对照组差异显著(P< 0.05),表明CAT和GPx基因参与了低盐胁迫反应,并表现出一定时间效应关系.综合分析结果指出脊尾白虾CAT基因是CAT家族主要成员之一,可能在低盐环境胁迫过程中具有重要的作用. |
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