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大黄鱼( Larimichthys crocea ) CRFB13 基因的克隆与表达分析
杨思司,母尹楠,丁连帅,陈新华,敖敬群
0
(国家海洋局第三海洋研究所、国家海洋局海洋生物遗传资源重点实验室, 福建 厦门 361005)
摘要:
IFN-γ是哺乳动物的一个关键免疫调节因子,通过特异性地结合IFN-γ受体1 (IFNGR1)和IFN-γ受体2 (IFNGR2) 组成的异源四聚受体复合物,来发挥其生物学功能.目前,在鱼类中已经克隆出IFNGR1基因的2个亚型:细胞因子受体家族B 13( CRFB13 )和细胞因子受体家族B17( CRFB17 ).本研究从大黄鱼( Larimichthys crocea )中克隆得到了一个 CRFB13 基因( LycCRFB13 ),其开放阅读框全长1 158个核苷酸,编码由385个氨基酸组成的1个蛋白.LycCRFB13蛋白具有典型的II型细胞因子受体特征,包括1个信号肽、1个胞外的FN III-like 结构域,1个单次跨膜结构域,以及胞内的JAK1和STAT1结合位点.氨基酸序列分析表明鱼类的CRFB13都具有JAK1 和 STAT1结合位点.系统进化分析表明鱼类的2种IFNGR1,CRFB13和CRFB17形成2个独立的分支,LycCRFB13和鱼类的CRFB13聚在一起,与雀鲷( Stegastes partitus )CRFB13的亲缘关系最近.组织分布显示 LycCRFB13 为组成型表达,在大黄鱼鳃中的表达量最高,而在小肠中的表达量最低.Poly(I:C)刺激后,大黄鱼鳃和头肾中 CRFB13 的mRNA转录水平都显著升高.此外,poly(I:C)和LPS还能够诱导大黄鱼头肾白细胞中 LycCRFB13 的表达,这些结果表明LycCRFB13可能在大黄鱼抗病毒、抗细菌免疫反应中发挥重要作用.
关键词:  海洋生物学  大黄鱼  IFN-γ受体  CRFB13  基因克隆  表达分析
DOI:
基金项目:国家自然科学基金资助项目(31372556)
Cloning of large yellow croaker ( Larimichthys crocea ) CRFB13 gene and expression analysis
YANG Si-si,MU Yin-nan,DING Lian-shuai,CHEN Xin-hua,AO Jing-qun
(Key Laboratory of Marine Biogenetic Resources, Third Institute of Oceanography, SOA,Xiamen 361005, China)
Abstract:
Interferon gamma (IFN-γ) is a major immunomodulatory factor in mammals. IFN-γmediates its effects by binding a tetrameric receptor complex which is comprises of two IFN-γ receptor (IFNGR) 1 chains and two IFNGR2 chains. Two isoforms of IFNGR1, cytokine receptor family member B (CRFB) 13 and CRFB17, have been isolated from some teleost fish. In this study, a CRFB13 homologue ( LycCRFB13 ) was identified and characterized in large yellow croaker ( Larimichthys crocea ). The open reading frame (ORF) of LycCRFB13 is 1 158 bp, encoding a protein consisting of 385 amino acids. The deduced LycCRFB13 protein exhibits a typical domain architecture of type II cytokine receptors, including a signal peptide, an extracellular FN III-like domain, a transmembrane region and intracellular JAK1 and STAT1 docking sites. Amino acid sequences alignment also revealed that all fish CRFB13 have potential JAK1 and STAT1 docking sites. Phylogenetic analysis showed that LycCRFB13 is closely related to CRFB13 of damselfish ( Stegastes partitus ) and falls into a clade formed by all fish CRFB13 sequences, separated from CRFB17 clade. LycCRFB13 mRNA was constitutively expressed in all tissues tested, with higher expression level in gills and lower in intestine. The transcription levels of LycCRFB13 mRNA were significantly up regulated in gills and head kidney after stimulated by poly(I:C). Moreover, both LPS and poly(I:C) also could induce expression of LycCRFB13 in head kidney Leukocytes of large yellow croaker. In all, the results indicate that LycCRFB13 may play roles in antiviral and antibacterial immune responses in large yellow croaker.
Key words:  marine biology  large yellow croaker ( Larimichthys crocea )  interferon-γ receptor  CRFB13  gene cloning  expression analysis

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