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鹅2'-5'-寡腺苷酸合成酶样基因的鉴定:基因结构、表达模式及对新城疫病毒的抗病毒活性

Identification of 2'-5'-Oligoadenylate Synthetase-Like Gene in Goose: Gene Structure, Expression Patterns, and Antiviral Activity Against Newcastle Disease Virus.

作者信息

Yang Chao, Liu Fei, Chen Shun, Wang Mingshu, Jia Renyong, Zhu Dekang, Liu Mafeng, Sun Kunfeng, Yang Qiao, Wu Ying, Chen Xiaoyue, Cheng Anchun

机构信息

1 Institute of Preventive Veterinary Medicine, Sichuan Agricultural University , Chengdu, China .

2 Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University , Chengdu, China .

出版信息

J Interferon Cytokine Res. 2016 Sep;36(9):563-72. doi: 10.1089/jir.2015.0167.

Abstract

2'-5'-oligoadenylate synthetase-like (OASL) is a kind of antiviral protein induced by interferons (IFNs), which plays an important role in the IFNs-mediated antiviral signaling pathway. In this study, we cloned and identified OASL in the Chinese goose for the first time. Goose 2'-5'-oligoadenylate synthetase-like (goOASL), including an ORF of 1527bp, encoding a protein of 508 amino acids. GoOASL protein contains 3 conserved motifs: nucleotidyltransferase (NTase) domain, 2'-5'-oligoadenylate synthetase (OAS) domain, and 2 ubiquitin-like (UBL) repeats. The tissue distribution profile of goOASL in 2-week-old gosling and adult goose were identified by Real-Time quantitative PCR, which revealed that the highest level of goOASL mRNA transcription was detected in the blood of adult goose and gosling. The mRNA transcription level of goOASL was upregulated in all tested tissues of duck Tembusu virus (DTMUV)-infected 3-day-old goslings, compared with control groups. Furthermore, using the stimulus Poly(I: C), ODN2006, R848, and lipopolysaccharide (LPS) as well as the viral pathogens DTMUV, H9N2 avian influenza virus (AIV), and gosling plague virus (GPV) to treat goose peripheral blood mononuclear cells (PBMCs) for 6 h, goOASL transcripts level was significantly upregulated in all treated groups. To further investigate the antiviral activity of goOASL, pcDNA3.1(+)-goOASL-His plasmid was constructed, and goOASL was expressed by the goose embryo fibroblast cells (GEFs) transfected with pcDNA3.1(+)-goOASL-His. Our research data suggested that Newcastle disease virus (NDV) replication (viral copies and viral titer) in GEFs was significantly reduced by the overexpression of goOASL protein. These data were meaningful for the antiviral immunity research of goose and shed light on the future prevention of NDV in fowl.

摘要

2'-5'-寡腺苷酸合成酶样蛋白(OASL)是一种由干扰素(IFN)诱导产生的抗病毒蛋白,在IFN介导的抗病毒信号通路中发挥重要作用。在本研究中,我们首次在中国鹅中克隆并鉴定了OASL。鹅2'-5'-寡腺苷酸合成酶样蛋白(goOASL),包含一个1527bp的开放阅读框,编码一个由508个氨基酸组成的蛋白质。GoOASL蛋白包含3个保守基序:核苷酸转移酶(NTase)结构域、2'-5'-寡腺苷酸合成酶(OAS)结构域和2个泛素样(UBL)重复序列。通过实时定量PCR鉴定了goOASL在2周龄雏鹅和成年鹅中的组织分布情况,结果显示在成年鹅和雏鹅的血液中检测到goOASL mRNA转录水平最高。与对照组相比,鸭坦布苏病毒(DTMUV)感染的3日龄雏鹅所有检测组织中goOASL的mRNA转录水平均上调。此外,使用多聚肌苷酸:多聚胞苷酸(Poly(I:C))、ODN2006、R848和脂多糖(LPS)以及病毒病原体DTMUV、H9N2禽流感病毒(AIV)和小鹅瘟病毒(GPV)处理鹅外周血单个核细胞(PBMCs)6小时,所有处理组中goOASL转录本水平均显著上调。为进一步研究goOASL的抗病毒活性,构建了pcDNA3.1(+)-goOASL-His质粒,并通过用pcDNA3.1(+)-goOASL-His转染的鹅胚成纤维细胞(GEFs)表达goOASL。我们的研究数据表明,goOASL蛋白的过表达显著降低了GEFs中新城疫病毒(NDV)的复制(病毒拷贝数和病毒滴度)。这些数据对鹅的抗病毒免疫研究具有重要意义,并为未来家禽NDV的预防提供了线索。

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