红笛鲷IRAK-4基因cDNA全长的克隆及组织表达分析  

Full-length c DNA Cloning and Tissue Expression Analysis of IRAK-4 Gene from Humphead Snapper(Lutjanus sanguineus)

作　　者：黄郁葱[1,2,3,4] 鲁义善[3,4] 简纪常[3,4] 吴灶和[4,5] 

机构地区：[1]中国科学院南海海洋研究所,广东广州510301 [2]中国科学院大学,北京100049 [3]广东海洋大学水产学院,广东湛江524088 [4]广东省水产经济动物病原生物学及流行病学重点实验室,广东湛江524088 [5]仲恺农业工程学院,广东广州510225

出　　处：《广东海洋大学学报》2015年第1期18-27,共10页Journal of Guangdong Ocean University

基　　金：十二五国家科技支撑计划(2012BAD17B03);广东省海洋经济创新发展区域示范专项(GD2012-B01-004);国家自然基金(41240041)

摘　　要：白细胞介素-1受体相关激酶4（interleukin-1 receptor-associated kinase 4,IRAK-4）是一种参与机体先天性免疫和适应性免疫反应过程中的关键分子。采用RT-PCR和c DNA末端快速扩增（RACE-PCR）的方法从红笛鲷（Lutjanus sanguineus）头肾中克隆IRAK-4基因的c DNA全序列（登录号：KF279357）。该序列全长2 015 bp,包含5′非编码区（5′UTR）205 bp,3′非编码区（3′UTR）421 bp,开放阅读框（ORF）1 389 bp,编码462个氨基酸。根据推导的氨基酸序列预测其蛋白分子质量为52.0 ku,理论等电点为5.19。氨基酸序列比对结果显示,红笛鲷IRKA-4基因氨基酸序列与其他物种的同源性为54.2%~85.7%。系统进化分析结果显示,红笛鲷与斜带石斑鱼（Epinephelus coioides）聚为一支,两者有较近的亲缘关系。用荧光定量PCR分析红笛鲷基因的组织表达差异,红笛鲷IRKA-4基因在各组织中均有不同程度的表达,其中在皮肤、肝脏和胃中表达量最高,其次是胸腺、鳃、心脏、肠、肌肉和脾脏,在头肾、后肾和脑的表达量最低。Interleukin-1 receptor-associated kinase 4（IRAK-4） is a key molecular which participates in the innate immune and adaptive immune processes. In this paper, full length c DNA sequence of IRAK-4 gene was amplified by RT-PCR and RACE PCR from head kidney of humphead snapper, Lutjanus sanguineus（Gen Bank accession number： KF279357）. The total c DNA sequence of humphead snapper IRAK-4 was 2 015 bp, including 3＇ UTR of 421 bp,5＇ UTR of 205 bp,an open reading frame（ORF） of 1 389 bp encoding 462 amino acids with Molecule Mass of 52.0 ku and p I of 5.19. The deduced amino acid sequence of humphead snapper IRAK-4 shared 54.2%- 85.7% identities with other species IRAK-4. Phylogenetic analysis showed that humphead snapper was clustered closely with orange-spotted grouper. In addition, the m RNA expression levels in different tissues were analyzed by real time quantitative PCR. The result showed that humphead snapper IRAK-4 expressed in all examined tissues with highest levels in skin, liver and stomach, moderate levels in thymus, gill, heart, intestine, muscle and spleen, and lowest levels in head kidney, kidney and brain.

关 键 词：红笛鲷 IRAK-4 基因克隆 组织表达 

分 类 号：Q78[生物学—分子生物学] Q959.483