池蝶蚌IRAK4基因的克隆及功能  

Molecular structure characteristics and function analysis of HsIRAK4 in Hyriopsis schlegelii

作　　者：周叶 王小敏 洪一江[1,2] ZHOU Ye;WANG Xiaomin;HU Beijuan;HONG Yijiang(School of Life Sciences,Nanchang University,Nanchang 330031;Key Laboratory of Aquatic Animal Resources and Utilization of Jiangxi,Nanchang University,Nanchang 330031,China)

机构地区：[1]南昌大学生命科学学院,南昌330031 [2]江西省水产动物资源与利用重点实验室,南昌330031

出　　处：《南昌大学学报（理科版）》2021年第4期383-393,共11页Journal of Nanchang University(Natural Science)

基　　金：科技部国家重点研发计划(2018YFD0901400);国家自然科学基金资助项目(31660337&31860737);国家现代农业产业技术体系(CARS-49)。

摘　　要：在优质淡水珍珠育珠贝池蝶蚌(Hyriopsis schlegelii)中获得白细胞介素受体相关激酶IRAK4基因cDNA全长序列,命名为HsIRAK4,其全长为2617 bp,5’端非翻译区(5′UTR)为146 bp,3′端非翻译区(3′UTR)为797 bp,ORF框1674 bp,编码557个氨基酸,包含2个保守结构域,即N端的死亡结构域(Death,DD)和激酶结构域(KD)。系统进化树分析显示,HsIRAK4与青蛤的同源性最高,与软体动物聚在一支,和鱼类、灵长类不在一支。采用Real time-quantitative PCR技术检测脂多糖LPS诱导后的HsIRAK4及其在TLR信号通路中的上游因子HsMyD88的mRNA表达水平,结果显示,HsMyD88和HsIRAK4在所有组织都有表达,并且HsMyD88和HsIRAK4都在肝胰腺、鳃、肾和肠等免疫相关组织中高表达,二者在LPS刺激后不同组织达到峰值的时间明显存在差异,说明HsMyD88和HsIRAK4很有可能参与贝类动物的免疫防御。池蝶蚌GST-Pulldown结果显示,HsMyD88的Death结构域分别能和HsIRAK4-Death和HsIRAK4-ORF发生相互作用。In this study,for the first time,the full-length cDNA sequence of interleukin receptor-related kinase IRAK4 gene was obtained in Hyriopsis schlegelii.The full-length cDNA sequence was named HsIRAK4,which was 2617 bp in length.The length 5′UTR of HsIRAK4 was146 bp.The length 3′UTR of HsIRAK4 was797 bp.The length ORF of HsIRAK4 was1674bp,encoding 557 amino acids,contains two conserved domains,the N-terminal death domain(Death,DD),and the kinase domain(KD).Phylogenetic tree analysis shows that HsIRAK4 has the highest homology with Cyclina sinensis,clustered with mollusks,and not with fish and primates.Real time-quantitative PCR was used to detect the mRNA expression level of HsIRAK4 and HsMyD88 induced by lipopolysaccharide LPS.The results showed that HsMyD88 and HsIRAK4 were expressed in all tissues,and HsMyD88 and HsIRAK4 were highly expressed in hepatopancreas,gill,kidney,intestine and other immune-related tissues.There were significant differences in the peak time of the two in different tissues after LPS stimulation,indicating that HsMyD88 and HsIRAK4 are likely to participate in the immune defense of shellfish.GST-Pulldown results show that the Death domain of HsMyD88 can interact with HsIRAK4-Death and HsIRAK4-ORF respectively.

关 键 词：池蝶蚌 HsIRAK4 HsMyD88 脂多糖LPS 荧光定量PCR GST-Pulldown 

分 类 号：Q959.223[生物学—动物学]