【摘要】 哺乳动物中, TRAF家族相关的NF-κB激活因子(TRAF family member-associated NF-κB activator, TANK)参与调控核因子κB (nuclear factor-κB, NF-κB)信号通路,在硬骨鱼中,最新研究表明TANK在青鱼抗病毒天然免疫反应中具有重要的功能,但在模式动物斑马鱼(Danio rerio)中还没有有关TANK的报道。本研究率先在斑马鱼开展TANK功能研究,克隆并获得了斑马鱼TANK基因(Danio rerio TANK, DrTANK),其编码区(coding sequence, CDS)包括1 047个核苷酸,编码349个氨基酸。免疫印迹实验显示Dr TANK蛋白大小约50 kD;免疫荧光实验证明Dr TANK主要定位于细胞质中,表明其为一种胞质蛋白;双荧光素酶报告基因研究结果显示,在EPC细胞中,过表达Dr TANK并不能显著上调干扰素启动子的表达,但聚肌胞苷酸[poly (I:C)]和草鱼呼肠孤病毒(grass carp reovirus, GCRV)的刺激能显著增强其诱导干扰素启动子的表达活性。本研究结果表明Dr TANK在斑马鱼抗病毒天然免疫反应中可能具有重要作用,为后续Dr TANK功能研究奠定了基础。更多还原

【Abstract】 In mammals, the TRAF family member-associated NF-κB activator(TANK) is involved in the regulation of the nuclear factor-κB(NF-κB) signaling pathway. Recent studies have showed that TANK plays an important role in the antiviral innate immune activation of black carp. However, in one of the most widely used animal model zebrafish, there is no report on its function. Herein, zebrafish(Danio rerio) TANK(DrTANK)was cloned and characterized. The DrTANK comprises 1 047 nucleotides and the predicted protein contains349 amino acids. The DrTANK migrated to approximately the 50 kD position in immunoblotting assay. The DrTANK was identified as a cytoplasmic protein by immunofluorescence staining. DrTANK could not induce the activity of interferon(IFN) promoter in luciferase reporter assay in EPC cells; however, the IFN-activation ability of DrTANK was obviously enhanced when the cells were treated with poly(I:C) or grass carp reovirus(GCRV). In summary, our data suggest that the DrTANK may play an important role in zebrafish antiviral in-nate immune response. This result benefits the functional study of the DrTANK in future.更多还原