大鳍鳠IL-1β基因cDNA克隆及表达分析  

Molecular cloning and expression analysis of the IL-1β gene in Mystus macropterus

作　　者：杨莹莹[1] 张其中[1,2] 李春涛[1] 陈霞[1] 朱成科[1] 李超[1] 

机构地区：[1]西南大学生命科学学院,淡水生物生殖与发育教育部重点实验室,三峡库区生态环境教育部重点实验室,水产科学重庆市市级重点实验室,重庆400715 [2]暨南大学水生生物研究所,广东广州510632

出　　处：《水产学报》2012年第3期321-328,共8页Journal of Fisheries of China

基　　金：国家“八六三”高技术研究发展计划(2011AA10A216);重庆市科技攻关计划项目(CSTC.1010AC1116);三峡库区生态环境教育部重点实验室基金资助项目

摘　　要：白细胞介素1β(interleukin-1β,IL-1β)作为炎症反应的关键介导物,通过开启基因的表达来调控免疫反应。为了丰富鱼类该基因的研究,实验采用RT-PCR和RACE-PCR技术,首次从鲇形目鱼类大鳍鳠中获得IL-1β基因cDNA的全序列。大鳍鳠IL-1β基因cDNA全长1 194 bp,包括3'非编码区域(UTR)为224 bp,5'UTR为82 bp,开放阅读框(ORF)为888 bp,编码296个氨基酸。预测的蛋白质分子量为33.843 43 ku,等电点为5.00。与斑点叉尾等其他8种脊椎动物进行同源性分析发现,大鳍鳠IL-1β氨基酸序列与斑点叉尾相似性最高,为79.0%,与原鸡的相似性最低,为22.0%。进化树分析表明,大鳍鳠IL-1β与斑点叉尾聚为一支。半定量PCR显示,大鳍鳠IL-1β基因在肌肉、脑、脾脏、头肾、体肾、胃、皮肤、肝脏及肠等组织中都有不同程度的表达;经嗜水气单胞菌刺激后大鳍鳠IL-1β基因在头肾和脾脏的转录表达显示,1 d后该基因在头肾与脾脏中的表达量都达到最大,15 d后恢复到刺激前的水平。研究显示,大鳍鳠IL-1β在抵御病原感染过程中发挥重要作用。Interleukin-1β（IL-1β）is a key mediator of inflammation and orchestrates a variety of immune responses by initiating gene expression.IL-1β was not identified from non-mammalian species until recently.However,no IL-1β genes have been reported from Siluriformes fishes class in China.In this study,we have cloned and sequenced the IL-1β in Mystus macropterus by RT-PCR and RACE-PCR methods.It contains 1 194 nucleotides,including 3′UTR of 224 nucleotides,5′UTR of 82 nucleotides,and an open reading frame with 888 nucleotides encoding a 296 amino acid peptide with MW of 33.84 ku and PI of 5.00.Compared with eight vertebrates species,IL-1β in M.macropterus shared the highest identity（79.0%）with that in Ictalurus punctatus,and the lowest identity（22.0%）with that in Gallus gallus.Phylogenetic tree analysis based on some IL-1β amino acids indicated that IL-1β amino acids in M.macropterus were clustered closely with Ictalurus punctatus.Expression analysis indicated that the IL-1β gene of M.macropterus was detected in muscle,brain,spleen,head kidney,kidney,stomach,skin,liver and intestine in varying degrees,and increased significantly in head kidney and spleen on 1 day after injection with Aeromonas hydrophila and returned to the normal level 15 d later.The result indicated that M.macropterus IL-1β plays a critical role in the host-pathogen interaction.

关 键 词：大鳍鳠 白细胞介素1Β 基因 克隆 组织分布 

分 类 号：S917[农业科学—水产科学] Q786[生物学—分子生物学]