真鲷去毒相关基因cDNA序列的克隆与肝脏组成型表达分析Molecular Cloning of Detoxification-Related Genes and Liver Constitutive Expression in Red Sea Bream Pagrus major
王琳;梁旭方;黄燕;胡永乐;陈亮;
摘要(Abstract):
赤潮发生时产生的一些海洋生物毒素对人类和海洋动物的安全形成潜在的威胁甚至导致死亡.为从分子水平探讨鱼类中海洋藻毒素的去毒分子机理,采用RT-PCR法克隆了真鲷(Pagrus major)肝脏芳香烃受体核转位蛋白(ARNT)和I时相异生素代谢酶细胞色素P4501A(CYP1A)基因cDNA核心序列,同时,应用半定量RT-PCR方法,以β-肌动蛋白作为外参照,在其指数期增长的范围内研究了芳香烃受体(AHR)、ARNT、CYP1A、II时相异生素代谢酶alpha型谷胱甘肽S-转移酶(GSTA1、GSTA2)、rho型谷胱甘肽S-转移酶(GSTR)、热休克蛋白70(HSP70)基因组成型表达水平.结果发现,真鲷ARNT、CYP1A基因cDNA核心序列片段分别长438bp和908bp,分别编码146和302个氨基酸.序列同源性分析发现,真鲷与门齿鲷(Stenotomus chrysops)、石首鱼(Micropogon undulatus)ARNT基因氨基酸序列同源性高达97.2%、95.2%,与斑马鱼(Brachydanio rerio)、人、大鼠、小鼠ARNT同源性较低(77.2%~79.3%).真鲷与门齿鲷、金头鲷(Sparus auratus)、欧洲川鲽(Rhombus maximus)、欧洲海鲈(Dicentrachus labrax)CYP1A基因氨基酸序列同源性较高,为84.8%~94.0%,与斑马鱼、人、小鼠CYP1A同源性较低,为59.6%~77.8%.真鲷肝脏AHR、ARNT、CYP1A、GSTA1、GSTA2、GSTR和HSP70基因组成型表达水平分别为(25.32±6.56)%、(26.22±4.24)%、(146.5±16.06)%、(55.42±3.75)%、(48.82±10.89)%、(79.47±3.13)%、(107.42±14.34)%.
关键词(KeyWords): 芳香烃受体(AHR);芳香烃受体核转位蛋白(ARNT);细胞色素P4501A;谷胱甘肽S-转移酶;热休克蛋白70;基因克隆;表达分析;真鲷(Pagrus major)
基金项目(Foundation): 国家科技部863项目(No.2007AA09Z437);; 国家自然科学基金项目(No.30670367);; 广东省科技计划项目(No.2007B020701002;No.2005B20301005);; 广东省自然科学基金项目(No.031886)
作者(Author): 王琳;梁旭方;黄燕;胡永乐;陈亮;
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