多鳞[鱼喜]fih-1基因克隆及其低氧胁迫的mRNA表达  

CLONING AND EXPRESSION ANALYSIS OF FIH-1 GENE IN SILLAGO SIHAMA UNDER HYPOXIA STRESS

作　　者：黄志棚 林星桦 王耀嵘 江东能 陈华谱[1,2,3] 邓思平 朱春华[1,2,3] 李广丽 黄洋[1,2,3] 田昌绪 HUANG Zhi-Peng;LIN Xing-Hua;WANG Yao-Rong;JIANG Dong-Neng;CHEN Hua-Pu;DENG Si-Ping;ZHU Chun-Hua;LI Guang-Li;HUANG Yang;TIAN Chang-Xu(Fisheries College,Guangdong Ocean University,Zhanjiang 524088,China;Guangdong Research Center on Reproductive Control and Breeding Technology of Indigenous Valuable Fish Species,Zhanjiang 524088,China;Guangdong Provincial Engineeing Laboratory for Maricwlture Organism Breeding,Zhanjiang 524088,China)

机构地区：[1]广东海洋大学水产学院,湛江524088 [2]广东省名特优鱼类生殖调控与繁育工程技术研究中心,湛江524088 [3]广东省海水养殖生物育种工程实验室,湛江524088

出　　处：《水生生物学报》2020年第4期744-748,共5页Acta Hydrobiologica Sinica

基　　金：广东省普通高校青年创新人才类项目(2018KQCX111);广东省普通高校特色创新类项目(2019KTSCX060);广东省科技计划项目(2014A020208117和2015A020209163);广东省大学生创新创业计划项目(CXXL2018048和CXXL2019138);广东海洋大学科研启动经费资助项目(R19026)资助。

摘　　要：为研究多鳞[鱼喜](Sillago sihama)低氧诱导因子抑制因子-1(factor inhibiting hypoxia inducible factor 1,fih-1)在低氧胁迫后的表达机制,实验通过PCR扩增技术克隆出多鳞[鱼喜]fih-1基因cDNA全长1280 bp,开放阅读框(ORF)1065 bp,编码353个氨基酸,具有JmjC保守结构域。多序列比对与聚类分析显示,多鳞[鱼喜]与大黄鱼和尖吻鲈的亲缘关系最近,与哺乳动物、两栖类和禽类的亲缘关系较远。fih-1基因mRNA在多鳞[鱼喜]多个组织中有不同表达,在精巢、卵巢和肌肉中的表达水平最高,在鳃、心脏和肝脏组织表达水平较高,在脑组织的表达水平较低。实时荧光定量PCR分析fih-1基因在低氧胁迫处理前后鳃和心脏组织中mRNA的表达情况,结果显示fih-1基因在两个组织均随低氧胁迫处理时间延长表达量显著升高(P<0.05)。研究表明fih-1基因在多鳞[鱼喜]低氧信号传导通路中扮演重要角色,为开展并解释多鳞[鱼喜]低氧胁迫遗传机制提供候选基因。Sillago sihama is a tropical fish from India to the western Pacific. It is widely distributed in the coastal waters of southern China and is one of the most important economic fish species. The factor inhibiting hypoxia inducible factor-1(fih-1) is a key factor in the hypoxia signal pathway. It plays an important role in the hypoxia adaptation in fishes. The fih-1 gene of S. sihama was cloned, and its cDNA sequence was 1280 bp, including an open reading frame of 1065 bp, encoding 353 amino. The structure prediction found that there was a JmjC conserved domain. Multi-sequence alignment and cluster analysis based on fih-1 amino acid sequences showed that the S. sihama was closely related to Larimichthys crocea and Lates calcarifer, bur farther to mammals, amphibians, and birds. The fih-1 was strongly expressed in the testis, ovary, and muscle, followed by the gill, heart and liver, but weakly expressed in the brain. The expression of fih-1 gene in gill and heart increased significantly after 1-hour hypoxia stress, and it continued to increase significantly after 6 hours. After 4 hours of normal dissolved oxygen, the expression decreased significantly but it was still higher than the normal level(P<0.05). This study revealed that fih-1 gene plays an important role in the hypoxic signal conduction pathway and provides candidate genes for carrying out and explaining the genetic mechanism of S. sihama.

关 键 词：多鳞[鱼喜] 低氧诱导因子抑制因子-1 低氧胁迫 克隆表达 序列分析 

分 类 号：Q344.1[生物学—遗传学]