中华鲟ASDIGIRR与ASTRAF6基因的克隆与表达

Cloning and expression of ASDIGIRR and ASTRAF6 genes of Acipenser sinensis

摘要:

中华鲟是处在硬骨鱼类与软骨鱼类之间的中国古老的珍稀鱼类之一，但在人工繁殖和养殖过程中，容易受到不同疾病的危害，因此，需要深入研究其免疫调控，为其疾病预防提供理论依据。单个免疫球蛋白白细胞介素-1受体相关分子(SIGIRR)和肿瘤坏死因子受体相关因子6(TRAF6)是toll样受体(TLR)信号通路的2个重要的信号转导元件。本研究鉴定了中华鲟的SIGIRR和TRAF6的同源物，分别命名为ASDIGIRR (Acipenser sinensis DIGIRR)和ASTRAF6 (A. sinensis TRAF6)，并研究它们在正常组织中的表达和Poly(I:C)诱导后的表达模式。结果显示，ASDIGIRR和ASTRAF6在健康鲟的10个组织中均有表达，且分别在肠道和头肾组织中表达量最高；用Poly (I:C)孵育中华鲟脾脏细胞后，ASDIGIRR在3 h时的表达量显著下调，在6 h时恢复至正常水平，后在24 h显著上调达到最高值，随后开始下调，至48 h时仍显著高于对照组，而ASTRAF6在6 h达到最高表达量，维持到24 h后再下调，48 h时下降至最低水平。研究表明，ASDIGIRR和ASTRAF6在中华鲟抵御病原入侵的免疫防御过程中起着重要作用。

关键词:中华鲟;ASDIGIRR;ASTRAF6;Poly (I:C);基因克隆;基因表达

Abstract:

Acipenser sinensis is one of the ancient and rare fishes in China between teleost fish and cartilaginous fish, but it is vulnerable to different diseases in the process of artificial reproduction and culture, so it is necessary to deeply study the immune regulation of A. sinensis to provide theoretical basis for its prevention of diseases. A single Ig-IL-1R-related molecule (SIGIRR) and tumor necrosis factor receptor-associated factor 6 (TRAF6) are two important signal transduction elements in the signal transduction pathway of toll-like receptors. In this study, homologues of SIGIRR and TRAF6 of A. sinensis, named ASDIGIRR and ASTRAF6, were identified, and their expression levels in normal tissues and Poly (I:C) induced expression were investigated. Results show that ASDIGIRR and ASTRAF6 were expressed in 10 tissues of healthy sturgeon, and the highest expression was found in intestinal tract and head kidney, respectively. After incubating A. sinensis spleen cells with Poly(I:C), the expression of ASDIGIRR was significantly down-regulated at 3 h, and returned to the normal level at 6 h, then increased to the highest level at 24 h and then decreased, and at 48 h still higher than the control group. While the expression of ASTRAF6 reached the highest level at 6 h, and maintained till 24 h, then decreased to the lowest level at 48 h. The results show that ASDIGIRR and ASTRAF6 play an important role in the immune defense of A. sinensis against pathogen invasion.

Key words:Acipenser sinensis;ASDIGIRR;ASTRAF6;Poly(I:C);gene cloning;gene expression