草鱼hepcidin基因cDNA克隆、序列分析与表达
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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)


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

    运用RT-PCR和快速扩增cDNA末端 (rapid amplication of cDNA 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 polymerase chain 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。

    Abstract:

    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 predicded 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 prohepcidin 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.

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韦友传,黄荣俊,陆专灵,陈代建,姬永杰,程光平.草鱼hepcidin基因cDNA克隆、序列分析与表达[J].上海海洋大学学报,2012,21(4):495-501.
WEI You-chuan, HUAN Rong-jun, LU Zhuan-ling, CHEN Dai-jian, JI Yong-jie, CHENG Guang-ping. Cloning, sequence analysis and expression of hepcidin in grass carp Ctenopharyngodon idellus[J]. Journal of Shanghai Ocean University,2012,21(4):495-501.

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  • 在线发布日期: 2012-07-18
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