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卵形鲳鲹(Pampus argenteus)MyD88 的克隆、鉴定及其对细菌刺激的反应功能。

Cloning, characterization, and function of MyD88 in silvery pomfret (Pampus argenteus) in response to bacterial challenge.

机构信息

Key Laboratory of Marine and Estuarine Fisheries, Ministry of Agriculture, East China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Shanghai 200090, People's Republic of China.

Key Laboratory of Marine and Estuarine Fisheries, Ministry of Agriculture, East China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Shanghai 200090, People's Republic of China.

出版信息

Int J Biol Macromol. 2017 Oct;103:327-337. doi: 10.1016/j.ijbiomac.2017.05.039. Epub 2017 May 11.

Abstract

Myeloid differentiation factor 88 (MyD88) is a key and universal downstream adapter for most Toll-like receptors (TLRs) and plays an important role in both the innate and adaptive immune response. In this study, the full-length cDNA of MyD88 (PAMyD88) from silvery pomfret (Pampus argenteus) was cloned and characterized. The PAMyD88 cDNA is 1545bp in length and contains an 876bp open reading frames (ORF). Multiple sequence alignment and phylogenetic tree analyzes revealed that the amino acid sequence of PAMyd88 was homologous to a variety of previously MyD88 molecules characterized from other species. The quantitative real-time reverse transcription-PCR analysis showed that the PAMyD88 mRNA was broadly expressed in all examined tissues, with higher levels observed in the immune-relevant organs. The results showed a significant up-regulation of the TLR2 and PAMyD88 transcript levels in response to L. plantarum and C. butyricum and a substantial expression level of TLR4 and PAMyD88 induced by V. anguillarum. Additionally, a challenge with V. anguillarum resulted in significant apoptosis, whereas the L. plantarum and C. butyricum induced only low levels of apoptosis. These data provide insight into the roles of PAMyD88 in the TLR signaling pathway in response to probiotic and pathogenic bacteria in silvery pomfrets.

摘要

髓样分化因子 88(MyD88)是大多数 Toll 样受体(TLR)的关键和通用下游衔接子,在先天和适应性免疫反应中都发挥着重要作用。在这项研究中,我们克隆并鉴定了银鲳(Pampus argenteus)MyD88(PAMyD88)的全长 cDNA。PAMyD88 cDNA 长 1545bp,包含一个 876bp 的开放阅读框(ORF)。多序列比对和系统发育树分析表明,PAMyd88 的氨基酸序列与多种先前从其他物种鉴定的 MyD88 分子同源。定量实时逆转录-PCR 分析显示,PAMyD88 mRNA 在所有检测的组织中广泛表达,在免疫相关器官中表达水平较高。结果表明,TLR2 和 PAMyD88 的转录水平在受到植物乳杆菌和丁酸梭菌刺激时显著上调,而 TLR4 和 PAMyD88 的表达水平在受到鳗弧菌刺激时显著上调。此外,鳗弧菌的攻击导致明显的细胞凋亡,而植物乳杆菌和丁酸梭菌仅诱导低水平的细胞凋亡。这些数据为 PAMyD88 在 TLR 信号通路中对益生菌和病原菌的反应中的作用提供了新的认识。

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