摘要
Toll样受体家族信号通路参与了机体特异性免疫和先天性免疫过程,白介素1受体相关激酶4 (IRAK4)是Toll样受体家族信号通路中的重要成员之一。该研究克隆了花鲈(Lateolabrax maculatus) lmirak4序列,并分析了其序列特征、表达模式及亚细胞定位。Lmirak4基因cDNA开放阅读框(ORF)全长942 bp,编码313个氨基酸,含有1个N端死亡结构域和1个中央蛋白激酶结构域。推导的Lmirak4氨基酸序列与其他鱼类Irak4氨基酸序列具有高度一致性。组织分布结果显示,lmirak4在各组织中表达具有差异性,在头肾中表达量最高。感染无乳链球菌(Streptococcus agalactiae)与哈维弧菌(Vibrio harveyi)后,花鲈头肾、肝脏和脾脏中lmirak4 mRNA的表达水平显著增加。亚细胞定位显示,外源性Lmirak4主要分布于HEK-293T细胞质中,在花鲈头肾原代培养细胞中成功表达但无明显分布差异。
Toll-like receptors(TLRs)play an indispensable role in the immune response of fish for pathogen recognition and triggering immune reaction,and interleukin-1 receptor-associated kinase 4(IRAK4)is one of the important components of TLR signaling pathway.We cloned the sequence of lmirak4 from Lateolabrax maculatus and analyzed its expression pattern and intracellular localization.The total cDNA sequence of lmirak4 ORF was 942 bp,encoding 313 amino acids with an N-terminal death domain and a central protein kinase domain.The Blast analysis shows that the deduced amino acid sequence of Lmirak4 shared high identity with other fishes.The tissue distribution analysis indicates that lmirak4 expressed in all the examined tissues with various expression levels and the highest expression was detected in head-kidney.Besides,the expression level of lmirak4 increased significantly in head-kidney,spleen and liver after infection of Vibrio harveyi and Streptococcus agalactiae.The eukaryotic expression plasmid of Lmirak4 was primarily expressed in the cytoplasm and membrane of HEK293T cells.In contrast,Lmirak4 showed a uniform presence of existence in L.maculatus primary kidney cells.
作者
李富祥
王鹏飞
闫路路
邱丽华
LI Fuxiang;WANG Pengfei;YAN Lulu;QIU Lihua(Key Laboratory of South China Sea Fishery Resources Exploitation&Utilization,Ministry of Agriculture and Rural Affairs;South China Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences,Guangzhou 510300,China;College of Fisheries and Life Science,Shanghai Ocean University,Shanghai 201306,China;Key Laboratory of Aquatic Genomics,Ministry of Agriculture and Rural Affairs,Beijing 100141,China)
出处
《南方水产科学》
CAS
CSCD
北大核心
2018年第5期70-79,共10页
South China Fisheries Science
基金
中国水产科学研究院中央级公益性科研院所基本科研业务费专项资金(2016PT04)
中国水产科学研究院南海水产研究所中央级公益性科研院所基本科研业务费专项资金资助(2016YD01,2017YB03,2017YB28,2018ZD01)
国家自然科学基金项目(31802281)
广东省自然科学基金项目(c18140500000602)。
关键词
花鲈
irak4
无乳链球菌
哈维弧菌
细胞转染
Lateolabrax maculatus
irak4
Vibrio harveyi
Streptococcus agalactiae
cell transfection
