团头鲂NOS2基因的克隆和组织表达分析  

Cloning and tissue expression analysis of inducible nitric oxide synthase gene from Megalobrama amblycephala

作　　者：王玉青 邬长松 马洁乐 陈会杰 刘小玲 WANG Yu-qing;WU Chang-song;MA Jie-le;CHEN Hui-jie;LIU Xiao-ling(Department of Aquatic Animal Medicine,College of Fisheries,Huazhong Agricultural University,Hubei Engineering Technology Research Center for Aquatic Animal Diseases Control and Prevention,National Demonstration Center for Experimental Aquaculture Education,Wuhan 430070,China)

机构地区：[1]华中农业大学水产学院水生动物医学系湖北省水生动物病害防控工程技术研究中心水产养殖国家级实验教学示范中心

出　　处：《淡水渔业》2019年第5期16-21,30,共7页Freshwater Fisheries

基　　金：国家自然科学基金(31772879);华中农业大学自主科技创新基金(2662017PY087)

摘　　要：为了探究诱导型一氧化氮合酶(NOS2)在嗜水气单胞菌(Aeromonas hydrophila)感染团头鲂(Megalobrama amblycephala)时发挥的作用,克隆并分析了团头鲂NOS2基因。结果显示,NOS2基因的cDNA序列全长4 215 bp,编码1 080个氨基酸。氨基酸序列同源性分析表明,团头鲂NOS2基因与草鱼NOS2基因的相似性最高,达95%。进化关系分析表明,团头鲂NOS2基因与鲤科鱼类NOS2基因聚在一支。实时荧光定量PCR检测的结果表明,在嗜水气单胞菌感染团头鲂后,NOS2基因在肝脏和头肾中被诱导表达;在脾脏中NOS2基因在3 h被诱导表达,其余时间点表达量显著下调;在肠道组织中NOS2基因在3 h下调表达,在6 h有微弱的上调,之后趋于稳定。研究结果为进一步探索NOS2基因在嗜水气单胞菌感染团头鲂后免疫应答中发挥的作用打下基础。Inducible nitric oxide synthase(NOS2)gene of Megalobrama amblycephala was cloned and characterized in order to study the function of inducible NOS2 gene from M.amblycephala after challenge of Aeromonas hydrophila.The full-length cDNA of NOS2 gene was 4 215 bp,encoding 1080 amino acids.Multiple sequence alignment revealed that NOS2 gene of M.amblycephala shared 95%similarity with that of Ctenopharyngodon idellus.Phylogenetic analysis showed that NOS2 amino acid sequence of M.amblycephala was relatively conserved in fish,and more closely related to Cypriniformes.The mRNA expression levels of the NOS2 gene were induced in the liver and head kidney after challenge of A.hydrophila.The expression in the spleen was up-regulated at 3h post injection(hpi).Subsequently,it was significantly down-regulated.The expression in the intestine was up-regulated at 6 hpi of A.hydrophila after a down-regulation at 3 hpi,and then tended to stabilization.Our findings will lay a foundation for the study of the potential role of the NOS2 gene in the host defense against A.hydrophila.

关 键 词：诱导型一氧化氮合酶(NOS2) 团头鲂(Megalobrama amblycephala) 嗜水气单胞菌(Aeromonas hydrophila) 组织表达 

分 类 号：S917.4[农业科学—水产科学]