大口黑鲈IRF3的分子特征及其免疫表达  

Molecular Characteristics of IRF3 Gene in Largemouth Bass(Micropterus salmoides)and Its Immune Expression

作　　者：荆鹏华 赵飞[2] 谭爱萍[2] 邓玉婷 邵蓬 朱雪晴 赖迎迢[2] 巩华[2] 黄志斌[2] JING Peng-hua;ZHAO Fei;TAN Ai-ping;DENG Yu-ting;SHAO Peng;ZHU Xue-qing;LAI Ying-tiao;GONG Hua;HUANG Zhi-bin(College of Fisheries,Tianjin Agricultural University,Tianjin 300392,China;Pearl River Fisheries Research Institute/Key Laboratory of Fishery Drug Development of Ministry of Agriculture and Rural Affairs/Key Laboratory of Aquatic Animal Immune Technology and Green Aquaculture of Guangdong Province,Pearl River Fisheries Research Institute,Chinese Academy of Fishery Sciences,Guangzhou 510380,China)

机构地区：[1]天津农学院水产学院,天津300392 [2]中国水产科学研究院珠江水产研究所/农业农村部渔用药物创制重点实验室/广东省水产动物免疫与绿色养殖重点实验室,广东广州510380

出　　处：《广东海洋大学学报》2023年第4期76-83,共8页Journal of Guangdong Ocean University

基　　金：中国水产科学研究院中央级公益性科研院所基本科研业务费专项(2022GH04,2021SJ-XT3,2020TD45);广州市农业和社会发展科技专题(2023B03J1305);佛山市南海区农业农村局政府采购项目(2967855)。

摘　　要：【目的】克隆大口黑鲈(Micropterus salmoides)干扰素调节因子3(interferon regulatory factor 3,IRF3)基因,命名为MsIRF3,探究该基因在大口黑鲈先天性免疫防御中的重要作用。【方法】根据大口黑鲈基因组中IRF3基因序列设计引物,克隆MsIRF3基因;用生物信息学方法分析MsIRF3的结构特征和理化性质;用鰤诺卡氏菌(Nocardia seriolae)或维氏气单胞菌(Aeromonas veronii)感染大口黑鲈,并用病原类似物脂多糖(LPS)、脂磷壁酸(LTA)和聚肌胞苷酸[Poly(I:C)]刺激大口黑鲈头肾白细胞,采用荧光定量PCR方法检测MsIRF3基因表达量的动态变化。【结果】MsIRF3的ORF全长为1404 bp,编码467个氨基酸。MsIRF3包含DNA结合结构域(DBD)、IRF关联结构域(IAD)及丝氨酸结构域(SRD)等3个典型结构域。同源性分析表明,MsIRF3与斜带石斑鱼(Epinephelus coioides)IRF3的亲缘性最近。qRT-PCR分析表明,MsIRF3在健康大口黑鲈各组织中广泛表达,在肠和鳃中表达量相对较高(P<0.05)。鰤诺卡氏菌或维氏气单胞菌感染后,MsIRF3在肝脏、头肾和脾中表达量均上调(P<0.01)。LPS、LTA和Poly(I:C)刺激后,大口黑鲈头肾白细胞中MsIRF3均被诱导表达。【结论】MsIRF3基因在大口黑鲈防御病原体感染的免疫应答中发挥重要作用。【Objective】To clone interferon regulatory factor 3 gene(IRF3)of largemouth bass(Micropterus salmoides),named MsIRF3,and explore its important function in the innate immune defense.【Method】Primers were designed based on the IRF3 gene from the genomic data of largemouth bass to clone the MsIRF3 gene.The structural characteristics and physicochemical properties were analyzed with bioinformatics methods.Quantitative real-time PCR(qRT-PCR)was used to detect the dynamic changes of MsIRF3 gene expression in largemouth bass tissues after infection with Nocardia seriolae or Aeromonas veronii,as well as the expression pattern of MsIRF3 in head kidney leukocytes after stimulation with three pathogenic analogs:lipopolysaccharides(LPS),lipoteichoic acid(LTA),and polyinosinic-polycytidylic acid[Poly(I:C)].【Result】The full-length ORF of MsIRF3 is 1404 bp in length,encoding a putative protein of 467 amino acids.MsIRF3 contains three typical domains:DNA binding domain(DBD),IRF association domain(IAD),and serine-rich domain(SRD).Homology analysis revealed that MsIRF3 was most closely related to IRF3 of Epinephelus coioides.The qRT-PCR analysis showed that MsIRF3 was commonly expressed in all the tested tissues of largemouth bass,with relatively higher expression in intestine and gills.After infection with N.seriolae and A.veronii,the expression of MsIRF3 was significantly up-regulated in the liver,head kidney,and spleen.Similarly,after stimulation with LPS,LTA,and Poly(I:C),the expression of MsIRF3 could also been induced in head kidney leukocytes.【Conclusion】MsIRF3 plays a crucial role in the immune response of largemouth bass to the pathogen infection.

关 键 词：大口黑鲈 干扰素调节基因3 鰤诺卡氏菌 维氏气单胞菌 免疫防御 

分 类 号：S917.4[农业科学—水产科学]