斑石鲷irf3基因鉴定及其在虹彩病毒感染下的表达模式
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Q 786;S 941.41

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国家重点研发计划 (2022YFF1000303);山东省重点研发计划 (2021LZGC028,2023ZLYS02);山东省良种工程项目 (2022LZGC016);中国水产科学研究院基本科研业务费专项 (2023TD20);山东省泰山学者攀登计划


Identification of irf3 gene in Oplegnathus punctatus and its expression pattern analysis under iridescent virus infection
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    摘要:

    干扰素调节因子 (irf3)是干扰素调节因子 (IRF)家族的一员,是I型干扰素依赖性免疫反应的主要转录调节因子,在针对DNA和RNA病毒的先天免疫反应中发挥重要作用。本研究中,斑石鲷irf3基因 (Oplegnathus punctatus interferon regulatory factors 3,Opirf3)来自实验室斑石鲷基因组数据库,经分析鉴定Opirf3的CDS序列全长1 362 bp,可编码453个氨基酸,预测其蛋白分子量为50.0 ku,理论等电点为4.97,有一个IRF结构域和一个IRF-3结构域。荧光定量分析结果显示,Opirf3在斑石鲷肝脏、鳃、心脏、皮肤、脾脏、肠、脑、肾脏、胃和头肾组织均有表达;虹彩病毒感染7 d时,免疫组织肝脏、脾脏和肾脏中Opirf3的表达水平显著上调。斑石鲷肾细胞系体外刺激实验显示,不同浓度poly I:C刺激后,Opirf3的表达量较对照组均显著升高,poly I:C浓度为100 μg/mL时,肾脏细胞中Opirf3的相对表达水平升高,为对照组的86.8倍。siRNA干扰后,斑石鲷肾细胞系中Opirf3表达水平显著下调30%,下游基因IFN-αCD40、CD80和IL-1β显著下调,IL-6显著上调。以上结果可能表明Opirf3基因参与了I型IFN在斑石鲷抗虹彩病毒过程中的先天免疫反应。本研究可为斑石鲷抗病分子育种提供理论依据。

    Abstract:

    Interferon regulatory factor 3, a member of the interferon regulator factor (IRF) family, are major transcriptional regulators of type I interferon-dependent immune responses and play an important role in innate immune responses against DNA and RNA viruses. In this study, The irf3 gene (interferon regulatory factors 3, Opirf3) was obtained from the Oplegnathus punctatus genome database, and analyses found that the CDS sequence of Opirf3 has a total length of 1 362 bp, encoding 453 amino acids. The molecular weight of predicted protein is 50.0 ku, the theoretical isoelectric point is 4.97, and has an IRF domain and an IRF-3 domain. RT-PCR showed that Opirf3 was expressed in liver, gills, heart, skin, spleen, intestine, brain, kidney, stomach and head kidney of O. punctatus. When the fish were infected with SKIV-SD for 7 days, the expression levels of Opirf3 in the liver, spleen and kidney of immune tissues were significantly upregulated. In vitro stimulation experiments of the kidney cell line of O. punctatus showed that the expression of Opirf3 was significantly upregulated after stimulation with different concentrations of poly I:C, and the relative expression level of Opirf3 was the highest at a concentration of 100 μg/mL, which was 87.9 times higher than that of the control group. After siRNA interference, the expression level of Opirf3 was significantly reduced by 30%, the Opirf3 related genes IFN-α, CD40, CD80 and IL-1β were significantly downregulated, and IL-6 was significantly upregulated. These results may indicate that the Opirf3 gene is involved in the innate immune response of type I IFN in the resistance to SKIV-SD of O. punctatus. This study can provide a theoretical basis for the molecular breeding of disease resistance of O. punctatus.

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李开敏,王磊,巩志宏,王清滨,李华,杨桂文,黄友华,陈松林.斑石鲷irf3基因鉴定及其在虹彩病毒感染下的表达模式[J].水产学报,2024,48(4):049428

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  • 收稿日期:2023-02-15
  • 最后修改日期:2023-06-09
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  • 在线发布日期: 2024-04-25
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