团头鲂谷胱甘肽S-转移酶基因的克隆及其在氨氮胁迫中的表达分析Molecular Cloning,Characterization and mRNA Expression of Mu-typ Glutathione S-Transferases from Megalobrama amblycephala
孙盛明;朱健;戈贤平;张成锋;缪凌鸿;张武肖;章琼;
摘要(Abstract):
谷胱甘肽S-转移酶(glutathione S-transferase,GST)是一类多功能蛋白家族,主要参与解毒和抗氧化防御过程。为了研究GST在团头鲂(Megalobrama amblycephala)肝脏解毒过程中的作用,克隆并分析了团头鲂1个谷胱甘肽S-转移酶基因(命名为MaGST)cDNA序列,采用实时荧光定量PCR研究了其在氨氮胁迫下的表达规律。MaGST包含1个长218个氨基酸的完整开放阅读框,具有GST蛋白家族的保守碱基和保守结构域。通过MEGA 5.0软件分析系统进化树发现,团头鲂GST与其他动物mu型GST聚为一簇,表明团头鲂GST属于mu型GST。荧光定量PCR结果显示MaGST基因在团头鲂各组织中均有表达,在肝脏和鳃中表达量最高,肌肉中表达量最低,同时在氨氮胁迫过程中该基因在肝和鳃中的表达规律相似,均在胁迫期间表达量显著上调;氨氮胁迫24 h时鳃和肝组织均存在组织损伤。研究结果提示在团头鲂肝脏和鳃组织中GST基因参与了氨氮胁迫的解毒过程。将该基因的编码区重组到p ET-21(a+)载体后在大肠杆菌中得到诱导表达,重组MaGST的GST活力为(10.36±0.68)U·mg~(-1)蛋白。
关键词(KeyWords): 氨氮胁迫;团头鲂;谷胱甘肽S-转移酶;基因克隆
基金项目(Foundation): 中央级公益性科研院所基本科研业务费专项资金(2015C06XK01);; 国家大宗淡水鱼类产业技术体系华东养殖岗位(CARS-46-14);; 十二五国家科技支撑计划“长江下游池塘高效生态养殖技术集成与示范”(2012BAD25B07)
作者(Author): 孙盛明;朱健;戈贤平;张成锋;缪凌鸿;张武肖;章琼;
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