赤眼鳟MyD88基因cDNA克隆与表达特性研究

MOLECULAR CLONING, CHARACTERIZATION AND EXPRESSION OF MyD88 IN SQUALIOBARBUS CURRICULUS

  • 摘要: 实验使用RACE-PCR技术获得了赤眼鳟(Squaliobarbus curriculus)髓样分化因子88 (Myeloid differentiationfactor 88, MyD88)的cDNA全长, 命名为ScMyD88。ScMyD88的cDNA全长为1779 bp, 其中开放阅读框855 bp, 共编码284个氨基酸残基, 推导的蛋白质分子量为33.053 kD, 理论等电点为5.66。赤眼鳟MyD88具有典型的MyD88结构特征, 包括死亡结构域和TIR结构域(Toll-IL-1 receptor domain, TIR), 其氨基酸序列和鲤科鱼类具有高度保守性, 相似性达到了90%以上, 特别是和武昌鱼相比, 相似性达到了98%。在检测的9个赤眼鳟组织和器官中均有MyD88表达, 其中肝脏、头肾和体肾中表达水平最高, 在脑中表达量最低。在草鱼呼肠弧病毒(Grass carp reovirus, GCRV)感染初期(12h内), MyD88在赤眼鳟免疫组织中表达水平急剧上升, 特别是在脾脏和体肾中尤为明显, 随后恢复正常水平。研究表明, MyD88在赤眼鳟抵抗GCRV入侵的免疫应答反应初期发挥了重要作用。

     

    Abstract: The full cDNA sequence of Myeloid differentiation factor 88 (MyD88) in the barbel chub Squaliobarbus curriculus (designated as ScMyD88) has been cloned by using RACE-PCRs method. The complete sequence of ScMyD88 cDNA was 1779 bp that contained 855 bp open read frame encoding 284 amino acid residues with calculated molecular weight of 33.053 kD and a theoretical isoelectric point (PI) of 5.66. The ScMyD88 protein has typical structural features of the MyD88 family with a Death domains and a TIR domain. According to the comparison with known MyD88 amino acid sequences, the putative protein of ScMyD88 showed the highest identities with cyprinid fishes (90%98%) and other bony fishes (67%74%) but low identities (60%) with mammalian. MyD88 expressed in all the nine tested tissues and organs with highly expressed in the liver, head kidney and trunk kidney, and lowly expressed in the brain. The ScMyD88 was obviously up-regulated in the immune tissue at 12h and returned to the normal level after 48h challenged with GCRV. These result showed that ScMyD88 might play important roles during the immune response of barbel chub against GCRV.

     

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