Molecular characterization and functional analysis of MyD88 in Chinese soft-shelled turtle Trionyx sinensis
Molecular characterization and functional analysis of MyD88 in Chinese soft-shelled turtle Trionyx sinensis
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DOI：
10.1016/j.fsi.2010.09.003
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发表时间：
2011-01-01
影响因子：
4.7
通讯作者：
Fang, Weihuan
中科院分区：
文献类型：
--
作者：
Li, Xiaoliang;Zhu, Binglin;Fang, Weihuan
髓样分化因子88 （MyD88）是触发先天免疫下游级联反应的信号转导的关键衔接蛋白之一。本研究首次从中华鳖（Trionyx sinensis）中鉴定出MyD88基因（tMyD88），这是爬行动物中首次鉴定出MyD88基因。tMyD88具有894 bp的ORF，编码297个氨基酸的多肽，包括n端典型的死亡结构域（DD）和c端保守的Toll/IL-1R （TIR）结构域。通过实时荧光定量PCR检测，该基因在嗜水气单胞菌活细胞攻毒的龟的脾脏、血液、肺和肝脏中表达量较高，但在肾脏和肠道中表达量较少。转染pcDNA-tMyD88的RAW 264.7细胞NF-kappa B活性高于载体对照（673.78 vs 410.72, P < 0.05）。转染pcDNA-tMyD88的RAW 264.7细胞中促炎因子IL-1 β和tnf - α的表达也显著高于转染pcDNA3.1的细胞（P < 0.01）。这些结果表明，tMyD88可能具有类似于哺乳动物的防御微生物感染的重要作用。(C) 2010年Elsevier Ltd.出版。
Myeloid differentiation factor 88 (MyD88) is one of the key adaptor proteins to signal transduction that triggers downstream cascades involved in innate immunity. In this study, the MyD88 gene from Chinese soft-shelled turtle (Trionyx sinensis) (tMyD88) was identified, representing the fist example from reptile species. The tMyD88 has a 894-bp ORF and encodes a polypeptide of 297 amino acids including a typical death domain (DD) at the N-terminus and a conservative Toll/IL-1R (TIR) domain at the C-terminus. It was expressed at high levels in spleen, blood, lungs and liver, but marginal in kidneys and intestines of turtles challenged with live cells of Aeromonas hydrophila, as determined by real-time PCR. RAW 264.7 cells transfected with pcDNA-tMyD88 showed higher NF-kappa B activity than the vector control (673.78 vs 410.72, P < 0.05). Expression of proinflammatory cytokines IL-1 beta and TNF-alpha was also significantly higher in RAW 264.7 cells transfected with pcDNA-tMyD88 than those having pcDNA3.1 control vector (P < 0.01). These results indicate that tMyD88 might possess an important role in defense against microbial infection in Chinese soft-shelled turtles similar to that in mammals. (C) 2010 Published by Elsevier Ltd.