草鱼hepcidin基因cDNA克隆、序列分析与表达  

Cloning,sequence analysis and expression of hepcidin in grass carp Ctenopharyngodon idellus

作　　者：韦友传[1] 黄荣俊[1] 陆专灵[1] 陈代建[1] 姬永杰[1] 程光平[1] 

机构地区：[1]广西大学动物科学技术学院,广西南宁530004

出　　处：《上海海洋大学学报》2012年第4期495-501,共7页Journal of Shanghai Ocean University

基　　金：广西大学科研基金(XBZ111496;XBZ111482);现代农业产业技术体系建设专项资金(GXLFY-02);科技部科技支撑计划项目(2008BADB9B05)

摘　　要：运用RT-PCR和快速扩增cDNA末端(rapid amplication ofcDNA Ends,RACE)技术获得草鱼(Ctenopharyngodon idellus)hepcidin基因全长cDNA,为774 bp,包括ORF 282 bp、5'UTR 117 bp和3'UTR 383 bp,3'UTR存在1个多聚腺苷酸加尾信号(AATAAA)和1个mRNA不稳定基序(ATTTA)。推定编码93个氨基酸,与其它鱼类hepcidin的序列同一性为27.9%～51.6%;SignalP 4.0软件预测信号肽位于1～24位。在邻接(neighbor-joining,NJ)法构建的系统进化树中,草鱼hepcidin前体肽和其他已报导的鱼类hepcidin前体肽聚为一枝。实时定量PCR(quantitative real-time polymerasechain reaction,qPCR)检测结果显示hepcidin基因mRNA主要表达于肝脏、脾脏、头肾和眼等组织;柱状黄杆菌注射后4～48 h,hepcidin基因在肝脏、脾脏和头肾中表达均显著上调。研究亮点:克隆到草鱼抗菌肽hepcidin一个新基因的全长cDNA,阐明了其所编码的hepcidin与其它脊椎动物hepcidin具有类似的结构与功能;证明其广泛分布于各组织中,参与了对细菌的免疫应答,是固有免疫的重要组成部分。The full cDNA sequence of hepcidin in grass carp Ctenopharyngodon idellus has been isolated by using RT-PCR and RACE method. The hepcidin consists of 774 bp, including an 282 bp open reading frame （ORF）, a 117 bp 5＇ UTR and a 383 bp 3＇ UTR . The 3＇ UTR contains a polyadenylation-tailed signal （AATAAA） and 1 mRNA instable motifs （ATTTA）. The deduced hepcidin protein comprised 93 amino acid residuces with a signal peptide predieded by using SignalP 4.0 program. Putative amino acid sequence of hepcidin shared 27.9% - 51.6% identity with its counterparts from other fish species. Phylogenetic tree constructed by using neighbor-joining method revealed that prohepeidin was clustered with that from other teleost fish reported previously. The transcription of hepcidin was examined by quantitative real-time PCR, and its mRNA was mainly expressed in liver, spleen and head kidney, and increased significantly in liver, spleen and head kidney between 4 h and 48 h after injection with Flavobacterium columnare.

关 键 词：草鱼 HEPCIDIN 表达 免疫应答 

分 类 号：S917.4[农业科学—水产科学] Q78[生物学—分子生物学]