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当前位置: 首页 > 文章 > 大鳍鳠IL-1β基因cDNA克隆及表达分析 水产学报 2012,36 (3) 321-328
Position: Home > Articles > Molecular cloning and expression analysis of the IL-1β gene in Mystus macropterus Journal of Fisheries of China 2012,36 (3) 321-328

大鳍鳠IL-1β基因cDNA克隆及表达分析

作  者：
杨莹莹;张其中;李春涛;陈霞;朱成科;李超
单  位：
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;暨南大学水生生物研究所
关键词：
大鳍鳠;白细胞介素1β;基因;克隆;组织分布
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。
译  名：
Molecular cloning and expression analysis of the IL-1β gene in Mystus macropterus
作  者：
YANG Ying-ying1,ZHANG Qi-zhong1,2,LI Chun-tao1,CHEN Xia1,ZHU Cheng-ke1,LI Chao1(1.Key Laboratory of Freshwater Fish Reproduction and Development,Key Laboratory of Eco-environments in Three Gorges Reservoir Region,Ministry of Education,Key Laboratory of Aquatic Science of Chongqing,School of Life Science,Southwest University,Chongqing 400715,China; 2.Hydrobiology Institute of Jinan University,Guangzhou 510632,China)
关键词：
Mystus macropterus;IL-1β;genes;clone;tissue distribution
摘  要：
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.

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