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鳗鲡髓样分化因子 88 基因的克隆与表达分析及其在鳗鲡抵御细菌感染中的作用

Molecular cloning and expression analysis of myd88 from oriental weatherfish (Misgurnus anguillicaudatus) in response to bacterial challenge.

机构信息

Jiangsu Key Laboratory of Marine Biotechnology/College of Marine Life and Fisheries, Jiangsu Ocean University, Lianyungang, China.

Fisheries Research Institute of Jiangxi Province, Nanchang, China.

出版信息

J Fish Biol. 2020 Jun;96(6):1341-1348. doi: 10.1111/jfb.14289. Epub 2020 Mar 12.

Abstract

Myeloid differentiation factor 88 (Myd88) plays an important role in both innate and adaptive immune response. In this study, the full-length complementary DNA (cDNA) of myd88 from Misgurnus anguillicaudatus was characterized. The myd88 cDNA is 1333 bp in length and contains an 855 bp open reading frame encoding a predicted protein of 284 amino acids. The predicted protein possesses typical Myd88 domain structural features including a death domain in the N-terminus, and box 1, 2, and 3 motifs of the Toll/IL-1 receptor domain in the C-terminus. Quantitative real-time PCR (qRT-PCR) revealed that myd88 messenger RNA (mRNA) was ubiquitously expressed in all examined tissues, especially highly in brain, kidney, blood, intestines and liver. qRT-PCR and western blotting were used to determine the mRNA and protein levels of Myd88 after Aeromonas veronii challenge, respectively. The Myd88 was remarkably upregulated in response to infection of A. veronii. These results suggested that Myd88 may play a vital role during the immune response of M. anguillicaudatus against bacterial infection.

摘要

髓样分化因子 88(Myd88)在先天和适应性免疫反应中都发挥着重要作用。本研究对泥鳅(Misgurnus anguillicaudatus)的 myd88 全长 cDNA 进行了鉴定。myd88 cDNA 全长 1333bp,包含一个编码 284 个氨基酸的预测蛋白的 855bp 开放阅读框。预测蛋白具有典型的 Myd88 结构域特征,包括 N 端的死亡结构域和 C 端 Toll/IL-1 受体结构域的框 1、2 和 3 基序。定量实时 PCR(qRT-PCR)显示,myd88 mRNA 在所有检测组织中广泛表达,尤其是在脑、肾、血、肠和肝中表达水平较高。qRT-PCR 和 Western blot 分别用于检测鳗弧菌感染后 Myd88 的 mRNA 和蛋白水平。Myd88 对鳗弧菌的感染反应显著上调。这些结果表明,Myd88 在泥鳅抵抗细菌感染的免疫反应中可能发挥重要作用。

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