日本鳗鲡TBK-1基因的克隆及免疫刺激的表达模式

Cloning and Expression Pofile Analysis After Immune Stimulation of TBK-1 Gene in Japanese Eel(Anguilla japonica)

利用PCR技术克隆了日本鳗鲡(Anguilla japonica)TBK-1基因(AjTBK-1),其开放阅读框为2193 bp,编码731个氨基酸。序列结构分析结果显示,AjTBK-1含有4个保守结构域,分别为氨基端激酶结构域,泛素样结构域和羧基端两个卷曲-螺旋结构域。系统发育分析表明,鱼类与四足类的TBK-1各自聚为一枝。实时定量PCR(qPCR)结果显示,AjTBK-1在日本鳗鲡各组织中均有表达。Poly I:C刺激6 h后,日本鳗鲡脾脏组织中AjTBK-1的上调倍数最高,为对照组的1.63倍;迟缓爱德华氏菌(Edwardsiella tarda)感染24 h后,日本鳗鲡肝脏组织中AjTBK-1的上调倍数最高,为对照组的2.2倍:表明AjTBK-1参与了日本鳗鲡抗病毒、抗细菌免疫反应应答。

The TBK-1 gene from Japanese eel(Anguilla japonica)named AjTBK-1 was cloned using polymerase chain reaction(PCR)technology,which has an open reading frame of 2193 bp encoding 731 amino acids.Sequence and structure analysis showed that AjTBK-1 contains four conserved domains,including an N-terminal kinase domain,a ubiquitin-like domain,and two coiled-coil domains at the C-terminus.Phylogenetic analysis indicated that fish and tetrapod TBK-1 formed separate branches.qPCR results showed that AjTBK-1 is expressed in various tissues of Japanese eel.After stimulation with Poly I:C for 6 h,the maximal expression of AjTBK-1 was detected in the spleen,which was 1.63-fold high than that of the control group.After infection with Edwardsiella tarda for 24 h,the maximal expression of AjTBK-1 was detected in the liver,which was 2.2-fold hign than that of the control group.Taken together,our results indicated that AjTBK-1 is involved in the antibacterial and antiviral immune responses of Japanese eel.