鲤鱼NOD1基因克隆与组织特异性表达分析

Molecular Cloning and Expression Analysis of NOD1 in Cyprinus Carpio

利用RT-PCR技术在鲤鱼(Cyprinus carpio)中首次克隆得到了NOD1基因编码序列全长。生物信息学分析发现,鲤鱼NOD1具有3个保守结构域和相应的保守位点,在进化中与草鱼亲缘关系最近;半定量RT-PCR分析NOD1基因在鲤鱼不同组织中分布发现,NOD1在头肾和血液中表达最高,在肌肉、皮肤和性腺中表达量最少;鳗弧菌(V.anguillarum)刺激后NOD1基因在肝脏、脾脏、头肾和前肠中表达量都有明显上升。研究结果显示NOD1在鲤鱼应对鳗弧菌刺激天然免疫过程中可能发挥着重要作用。

NOD1, a member of NLR family of pattern recognition receptors, plays a crucial role in the induction of defense reactions of the immune system against pathogenic bacterial and viral in infections. In this study, We firstly cloned ORF sequences of NOD1 gene in cyprinus carpio with RT-PCR. Three conserved domains and three conserved sites were identified by bioinformation analysis. Phyloge- netic analysis shows that the common carp NOD1 has the closest relationship with grass carp NOD1. Semi-quantitative RT-PCR analysis shows that common carp NOD1 gene is expressed at high level in head kidney and blood, but lower in muscle, skin and gonad. In vitro stimulation of V. anguillarum increases NOD1 gene expression in liver, spleen, head kidney and foregut. These results suggest taht NOD1 may play an important role in cyprinus carpio innate immune response.