Molecular characterization, expression patterns, and subcellular localization of RIG-I in the Jinding Duck (Anas platyrhynchos domesticus)
Molecular characterization, expression patterns, and subcellular localization of RIG-I in the Jinding Duck (Anas platyrhynchos domesticus)
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DOI：
10.1016/j.dci.2013.07.018
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发表时间：
2013-12-01
影响因子：
2.9
通讯作者：
Chen, Guohong
中科院分区：
文献类型：
--
作者：
Chen, Yang;Zhang, Yang;Chen, Guohong
视黄酸诱导基因I （RIG-I）样受体（RLRs）最近被鉴定为RNA病毒的细胞质传感器。最近的研究表明，rig - 1是该家族的一员，在先天免疫中起重要作用。本研究采用逆转录聚合酶链反应（RT-PCR）和cDNA末端快速扩增（RACE）技术克隆金顶鸭rig - 1基因。我们确定durig - 1的cDNA包含一个14bp的5' UTR，一个2802-bp的开放阅读框，以及两个3' UTR (295-bp和927-bp)，编码933个氨基酸的多肽。基于该序列，预测durig - 1蛋白具有典型rlr的保守结构域。此外，通过间接免疫荧光发现durig - 1分布在DF1细胞中，与预测一致。半定量RT-PCR （sqRT-PCR）在健康组织中几乎检测不到durig - 1 mRNA。为了研究rig - 1在先天免疫中的作用，我们利用合成的双链RNA模拟体内病毒感染，并通过实时荧光定量PCR （qRT-PCR）检测脾脏和肝脏中durig - 1转录物。在注射后8 h durig - 1 mRNA的表达显著升高（P < 0.05），与其他时间点的对照水平无明显差异（P < 0.05）。这些结果表明durig - 1在对双链RNA病毒的先天免疫应答中起重要作用，值得进一步研究以揭示其可能的机制。(C) 2013 Elsevier Ltd.版权所有。
Retinoic acid-inducible gene I (RIG-I)-like receptors (RLRs) have recently been identified as cytoplasmic sensors for RNA virus. Recent research has shown that RIG-I, a member of this family, play an important role in innate immunity. In this study, we cloned the RIG-I gene from Jinding duck by reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE). We determined that the cDNA of duRIG-I contains a 14-bp 5' UTR, a 2802-bp open reading frame, and alternative 3' UTRs (295-bp and 927-bp) and encodes a polypeptide of 933 amino acids. Based on this sequence, the duRIG-I protein is predicted to have conserved domains typical of RLRs. In addition, duRIG-I was found to be distributed throughout DF1 cells by indirect immunofluorescence, as predicted. duRIG-I mRNA was scarcely detected in healthy tissues by semi-quantitative RT-PCR (sqRT-PCR). To study the role of RIG-I in innate immunity, we used synthetic double-stranded RNA to mimic viral infection in vivo and detected duRIG-I transcripts in spleen and liver by quantitative real-time PCR (qRT-PCR). The expression of duRIG-I mRNA was significantly elevated at 8 h post-injection (P < 0.05) and was indistinguishable from control levels at other time points (P > 0.05). These results suggest that duRIG-I plays an important role in innate immune responses to double-stranded RNA viruses and warrant further studies to reveal the possible mechanism. (C) 2013 Elsevier Ltd. All rights reserved.