鳜IRAK4基因的克隆、组织表达及病毒感染后表达分析  

Cloning and expression profiling of ScIRAK4 gene in mandarin fish(Siniperca chuatsi)in response to virus infections

作　　者：林强[1,2] 杨淞[3] 李宁求[1,2] 方翔[1] 付小哲[1,2] 刘礼辉[1,2] 郭慧芝[1,2] 李万里[3] 吴淑勤[1,3] 

机构地区：[1]中国水产科学研究院珠江水产研究所,农业部渔用药物创制重点实验室,广东省水产动物免疫技术重点实验室,广东广州510380 [2]淡水水产健康养殖湖北省协同创新中心,湖北武汉430070 [3]四川农业大学动物科技学院,四川成都611130

出　　处：《水产学报》2016年第3期344-354,共11页Journal of Fisheries of China

基　　金：国家科技支撑计划(2012BAD25B02);国家自然科学基金(31502201);广东省海洋渔业科技与产业发展专项(A201501B12)~~

摘　　要：为了研究鳜IRAK4生物学特性及其在抗病毒免疫应答中的作用,根据鳜转录组数据中筛选出的IRAK4 unigene序列设计引物,利用SMART-RACE技术克隆得到CDS全长为1389 bp的c DNA(命名为Sc IRAK),编码462个氨基酸,含有1个N端死亡结构域和1个保守的中央蛋白激酶结构域。采用荧光定量RT-PCR方法分析了Sc IRAK4在健康鳜各组织中的表达差异及病毒感染后在脾脏中的表达变化,结果显示,健康鳜中Sc IRAK4在肝脏中表达量最大,与其他组织差异显著,而在血液、脑和胃中表达量最低;传染性脾肾坏死病毒(infectious spleen and kidney necrosis virus,ISKNV)感染鳜后Sc IRAK4的表达量呈现下调趋势,24 h脾脏中的表达量达到最低,为对照组的45%;而鳜弹状病毒(siniperca chuatsi rhabdovirus,SCRV)感染鳜后Sc IRAK4的表达量呈现上调趋势,12 h脾脏中Sc IRAK4的表达量达到最高,为对照组的8.17倍,表明Sc IRAK4在抗ISKNV和SCRV的免疫应答中可能发挥不同的作用。本研究为进一步揭示Sc IRAK4的抗病毒免疫反应机制提供了依据。As a pivotal signaling mediator of Toll-like receptor（TLR） and interleukin（IL）-1 receptor（IL-1R）signaling cascades, the IL-1R-associated kinase 4（IRAK4） is engaged in the activation of host immunity. To study its biological function in mandarin fish（Siniperca chuatsi）, the expression profiling of the gene and its role in immune responses to the infection of infectious spleen and kidney necrosis virus（ISKNV） and S. chuatsi Rhabdovirus（SCRV） were investigated. Based on the unigene sequences of IRKA4 gene which was obtained from the transcriptomic results of S. chuatsi, specific primers were designed and the complete IRKA4 gene（named Sc IRAK） was cloned and sequenced by SMART-RACE. Bioinformatics analysis demonstrated that the CDS of Sc IRAK4 gene was 1389 bp, encoding a 462 amino acid with an N-terminal death domain（DD） and a central protein kinase domain（PKc）. The transcription profiles of IRAK4 in the tissues of S. chuatsi were characterized by fluorescent quantitative RT-PCR. The results showed that the highest m RNA expression was found in the liver（P〈0.05）, followed by that in the muscle, blood, brain and stomach（P〈0.05）. The transcriptions of the IRAK4 in the spleen of mandarin fish infected with ISKNV or SCRV were furtherly analyzed. The m RNA of Sc IRAK4 was down-regulated in the mandarin fish infected with ISKNV and the lowest transcription was observed at 24 h post infection（P〈0.01）. By contrast, the m RNA of Sc IRAK4 was up-regulated in the mandarin fish infected with SCRV and the higest transcription was observed at 12 h post of the infection（P〈0.01）. These findings suggest that Sc IRAK4 plays a crucial and different role in the immune responses of Mandarin fish infected with different viruses.

关 键 词：鳜 SC IRAK4 基因克隆 组织表达 

分 类 号：Q785[生物学—分子生物学] S941[农业科学—水产养殖]