水产学报2024，Vol.48Issue(4)：346-357,12.DOI:10.11964/jfc.20230213910
斑石鲷irf3基因鉴定及其在虹彩病毒感染下的表达模式
Identification of irf3 gene in Oplegnathus punctatus and its expression pattern analysis under iridescent virus infection
摘要
Abstract
Interferon regulatory factor 3,a member of the interferon regulator factor(IRF)family,are major tran-scriptional 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 kid-ney 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 stimula-tion with different concentrations of poly I:C,and the relative expression level of Opirf3 was the highest at a con-centration 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.关键词
斑石鲷/irf3/基因表达/siRNAKey words
Oplegnathus punctatus/irf3/gene expression/siRNA分类
生物科学引用本文复制引用
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):346-357,12.基金项目
国家重点研发计划(2022YFF1000303) （2022YFF1000303）
山东省重点研发计划(2021LZGC028,2023ZLYS02) （2021LZGC028,2023ZLYS02）
山东省良种工程项目(2022LZGC016) （2022LZGC016）
中国水产科学研究院基本科研业务费专项(2023TD20) （2023TD20）
山东省泰山学者攀登计划 National Key R&D Program of China(2022YFF1000303) （2022YFF1000303）
Key Research and Development Project of Shandong Province(2021LZGC028,2023ZLYS02) （2021LZGC028,2023ZLYS02）
Shandong Province Breeding Project(2022LZGC016) （2022LZGC016）
Central Public-interest Scientific Institute Basal Research Fund,CAFS(2023TD20) （2023TD20）
Taishan Scholar Climbing Program of Shandong Province,China （）
