斜带石斑鱼干扰素调节因子8基因的鉴定与表达模式分析  

Identification and Expression Pattern Analysis of Interferon Regulatory Factor 8 Gene in Orange Spotted Grouper,Epinephelus coioides

作　　者：贾芹芹[1,2] 黄贝[1,2] 熊静[1,2] 段利朋 徐继松[1,2] 聂品[1] 黄文树[1,2,3] JIA Qinqin HUANG Bei(Fishery College,Jimei University, 2.Engineering Research Center of the Modern Technology for Eel Industry, Ministry of Education, Jimei University, 3.Fujian Collaborative Innovation Center for Exploitation and Utilization of Marine Biological Resources, Xiamen 361021,China)

机构地区：[1]集美大学水产学院 [2]鳗鲡现代产业技术教育部工程研究中心(集美大学) [3]福建省海洋生物资源开发利用协同创新中心,福建厦门361021

出　　处：《厦门大学学报（自然科学版）》2017年第2期178-187,共10页Journal of Xiamen University：Natural Science

基　　金：国家自然科学基金(U205123;31172438;31402329);国家高技术研究发展计划(2012AA092201);福建省自然科学基金(2012J06008;2014J05042);厦门南方海洋研究中心科技项目(14PYY050SF03)

摘　　要：干扰素调节因子8(IRF8)是IRF家族转录因子中的一个成员.利用简并PCR和cDNA末端快速克隆(RACE)技术获得了斜带石斑鱼(Epinephelus coioides)IRF8基因(EcIRF8)全长cDNA序列,其编码422个氨基酸残基的蛋白质,含有脊椎动物IRF家族的特征性结构域,即N端保守的DNA结合结构域(DBD)和C端多样化的干扰素调控因子结合结构域(IAD),其中DBD中包含5个特征性的色氨酸残基和1个核定位信号.与绝大多数脊椎动物的IRF8基因相似,EcIRF8基因组序列也是9个外显子和8个内含子的结构.EcIRF8在斜带石斑鱼肝脏中高水平表达.活体聚肌胞苷酸刺激后6h,EcIRF8在头肾、中肾、胸腺、肝脏和肠中的表达量显著上调(p<0.05),其中在中肾中上调幅度最大,是对照组的106倍;副溶血弧菌(Vibrio paraheamolyticus)刺激后24h,EcIRF8在肝脏中的表达量也显著上调(p<0.05),但上调倍数仅为对照组的9.37倍.结果表明,EcIRF8基因可能主要参与斜带石斑鱼的抗病毒免疫反应.Interferon regulatory factor 8 （IRF8） is a transcription factor in IRF family,and plays critical roles in the regulation of lineage commitment and in myeloidcell maturation including the decision for a common myeloid progenitor to differentiate into a monocyte precursor cell.In this study,an IRF8 gene was cloned from orange spotted grouper,Epinephelus coioides,using degenerate PCR and RACE. The full-length cDNA of EcIRF8 encodes 422 amino acid residues. The predicted peptide possesses two characteristic domains of IRF family,including a conserved DNA-binding domain with one nuclear localization signal and five conserved tryptophan residues in the N-terminal region,and a divergent C-terminal region that serves as the regulatory domain.EclRF8 shares a nine-exon/eight-intron structure with its vertebrate homologues, with one intron in 5＇-UTR. In normally cultured orangespotted grouper,EcIRF8 was highly expressed in liver.In vivo,the transcriptional expression of EclRF8 was significantly up-regulated in head kidney, middle kidney, thymus, liver and intestine, at 6 h post iniection with Poly I： C, and the maximum up-regulated expression of EclRF8 was in middle kidney, which reached up to 106 folds compared with the control.In addition, a 9.3-fold increasing expression of EcIRF8 was found in liver at 24 h post injection with Vibrio parahemolyticus. The results suggested that EclRF8 might be involved in both antiviral and antibacterial immune response of E. coioides,and mainly in defense from viral infection.

关 键 词：干扰素调节因子8 斜带石斑鱼 活体聚肌胞苷酸 

分 类 号：Q17[生物学—水生生物学]