傅明骏, 赵超, 杨其彬, 周发林, 江世贵, 邱丽华. 斑节对虾过氧化氢酶基因全长cDNA克隆及表达分析[J]. 南方水产科学, 2015, 11(6): 107-113. DOI: 10.3969/j.issn.2095-0780.2015.06.015
引用本文: 傅明骏, 赵超, 杨其彬, 周发林, 江世贵, 邱丽华. 斑节对虾过氧化氢酶基因全长cDNA克隆及表达分析[J]. 南方水产科学, 2015, 11(6): 107-113. DOI: 10.3969/j.issn.2095-0780.2015.06.015
FU Mingjun, ZHAO Chao, YANG Qibin, ZHOU Falin, JIANG Shigui, QIU Lihua. cDNA cloning and expression analysis of catalase from black tiger shrimp (Penaeus monodon)[J]. South China Fisheries Science, 2015, 11(6): 107-113. DOI: 10.3969/j.issn.2095-0780.2015.06.015
Citation: FU Mingjun, ZHAO Chao, YANG Qibin, ZHOU Falin, JIANG Shigui, QIU Lihua. cDNA cloning and expression analysis of catalase from black tiger shrimp (Penaeus monodon)[J]. South China Fisheries Science, 2015, 11(6): 107-113. DOI: 10.3969/j.issn.2095-0780.2015.06.015

斑节对虾过氧化氢酶基因全长cDNA克隆及表达分析

cDNA cloning and expression analysis of catalase from black tiger shrimp (Penaeus monodon)

  • 摘要: 过氧化氢酶(catalase,CAT)是一种重要的抗氧化酶,防止过氧化氢对机体的氧化应激。该研究以斑节对虾( Penaeus monodon) 肝胰腺转录组中筛选获得的CAT基因片段为基础,利用RACE 技术获得斑节对虾CAT基因(PmCAT)cDNA全长。该基因cDNA全长为1 909 bp,开放阅读框为1 563bp,编码520个氨基酸。PmCAT推导的氨基酸序列与其他动物氨基酸序列具有高度的一致性。实时定量PCR结果显示,PmCAT基因在斑节对虾的不同组织中均有表达,其中在鳃的表达量最高,肝胰腺和胸神经次之。在硫酸铜(CuSO4)胁迫下,PmCAT基因在肝胰腺中的表达量显著下调(P0.05),在第24和第48小时恢复正常表达量;在pH 7.0胁迫下,PmCAT表达量在第12小时升高达最大值(P0.05),然后恢复到正常水平;在pH 9.0胁迫下,PmCAT基因表达显著低于正常组(P0.05)。研究表明,PmCAT基因参与斑节对虾氧化应激和对环境毒理的适应性反应。

     

    Abstract: Catalase (CAT) is an antioxidant enzyme and plays an important role in protection against oxidative stress by scavenging hydrogen peroxide (H2O2). We obtained the full-length cDNA sequence ofCAT from black tiger shrimps ( Penaeus monodon) ( denoted as PmCAT) by transcriptome sequencing and RACE-PCR. The full-length cDNA of PmCAT was 1 909 bp,including an open reading frame ( ORF) of 1 563 bp encoding a polypeptide of 520 amino acids,a 5'UTR of 134 bp and a 3'UTR of 212 bp. Blast and phylogenetic analyses suggest that PmCAT is a CAT family member. The expression of PmCAT mRNA showed a declining trend under the stress of CuSO4 (P0.05), and then recovered to normal at member 24th hour and 48th hour, being the highest in gill, and then in hepatopancreas and thoracic nerves. Under the stress of pH 7.0,PmCAT expression reached the maximum at 12th hour (P0.05); under the stress of pH 9.0, the expression of PmCAT declined (P0.05). The results reveal that PmCAT plays a significant role in adaptive response to conditions of oxidative stress and environmental toxicity.

     

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