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Molecular cloning and functional analysis of duck ubiquitin-specific protease 18 (USP18) gene

机译:鸭泛素特异性蛋白酶18(USP18)基因的分子克隆和功能分析

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摘要

In mammals, ubiquitin-specific protease 18 (USP18) is an interferon (IFN)-inducible gene and is a negative regulator of Toll-like receptor-mediated nuclear factor kappa B (NF-kappa B) activation. The role of USP18 in ducks (duUSP18) remains poorly understood. In the present study, we cloned and characterized the full-length coding sequence of duUSP18 from duck embryo fibroblasts (DEFs). In healthy ducks, duUSP18 transcripts were broadly expressed in different tissues, with higher expression levels in the spleen, lung and kidney. Quantitative real-time PCR (qRT-PCR) analysis revealed that duUSP18 could be induced by treatment with Poly(I:C) or LPS. Overexpression of duUSP18 inhibited NF-kappa B and IFN-beta expression. Furthermore, deletion mutant analysis revealed that the duUSP18 region between as 75 and 304 was essential for inhibiting NF-kappa B. In addition, overexpression of duUSP18 also suppressed the secretion of NF-kappa B-dependent proinflammatory cytokines. Taken together, these results suggest that duUSP18 regulates duck innate immune responses. (C) 2016 Elsevier Ltd. All rights reserved.
机译:在哺乳动物中,泛素特异性蛋白酶18(USP18)是干扰素(IFN)诱导型基因,并且是Toll样受体介导的核因子κB(NF-κB)激活的负调节剂。 USP18在鸭子中的作用(duUSP18)仍然知之甚少。在本研究中,我们从鸭胚胎成纤维细胞(DEFs)中克隆并鉴定了duUSP18的全长编码序列。在健康的鸭子中,duUSP18转录本在不同组织中广泛表达,在脾脏,肺脏和肾脏中的表达水平更高。实时定量PCR(qRT-PCR)分析表明,duUSP18可以通过Poly(I:C)或LPS处理诱导。 duUSP18的过表达抑制了NF-κB和IFN-β的表达。此外,缺失突变体分析显示,duUSP18区域介于75和304之间对于抑制NF-κB是必不可少的。此外,duUSP18的过表达还抑制了NF-κB依赖性促炎细胞因子的分泌。两者合计,这些结果表明duUSP18调节鸭先天免疫反应。 (C)2016 Elsevier Ltd.保留所有权利。

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