鲤RNF14基因的生物学功能及其对病原刺激的免疫响应  

Biological function of RNF14 gene in Cyprinus carpio and its immune response to pathogen stimulation

作　　者：张美娜 张子豪 洪佳乐 郭朝辉 李海洁 于光晴 董鹏生 黄小城 杨振江 王延晖 刘变枝 ZHANG Meina;ZHANG Zihao;HONG Jiale;GUO Chaohui;LI Haijie;YU Guangqing;DONG Pengsheng;HUANG Xiaocheng;YANG Zhenjiang;WANG Yanhui;LIU Bianzhi(College of Animal Science and Technology,Henan Agricultural University,Zhengzhou 450046,China;Hualan Biological Engineering,INC,Xinxiang 453003,China;Laboratory of Aquaculture and Genetic Breeding,Henan Academy of Fishery Science,Zhengzhou 450044,China)

机构地区：[1]河南农业大学动物科技学院,河南郑州450046 [2]华兰生物工程股份有限公司,河南新乡453003 [3]河南省水产科学研究院水产养殖与遗传育种研究室,河南郑州450044

出　　处：《河南农业大学学报》2025年第2期255-265,共11页Journal of Henan Agricultural University

基　　金：国家自然科学基金项目(31702359);河南省科技研发联合基金项目(222103810017);河南省高等学校重点科研项目(23A240001);河南省现代农业产业技术体系(HARS-22-16-T)。

摘　　要：【目的】探究鲤环指蛋白14(ring finger protein 14,RNF14)基因的生物学功能,为阻断鲤春季病毒血症病毒(spring viremia of Cyprinus carpio virus, SVCV)感染提供新思路。【方法】采用PCR技术、生物信息学分析和实时荧光定量PCR方法,对鲤RNF14基因功能进行初步分析,研究其在病毒刺激后的表达规律。【结果】鲤RNF14基因编码区全长1 362 bp,编码453个氨基酸;鲤RNF14与犀角金线鲃亲缘关系较近,与原鸽亲缘关系较远;该蛋白属酸性不稳定亲水性蛋白;功能结构域预测显示该蛋白为典型的RBR E3s;泛素化底物分析结果显示,该蛋白与RBCK1、Nedd4l、Notch1a等蛋白相互作用的置信度较高。组织表达谱分析表明,鲤RNF14基因在各组织或器官中广泛表达,以心脏中相对表达量最高。与对照组相比,高剂量SVCV在体感染鲤24 h后,RNF14基因的mRNA表达水平在肌肉中显著升高;多肌胞苷酸(polyinosinic acid-polycytidylic acid,ploy(I:C))在体刺激鲤24 h后,RNF14基因的mRNA表达水平在肾脏和头肾中显著升高。与对照组相比,低剂量SVCV和poly(I:C)离体刺激下,鲤RNF14基因mRNA表达水平在各取样时间点均极显著升高。在体和离体病原刺激结果表明,鲤RNF14基因参与了病原刺激后的免疫应答过程,提示鲤RNF14基因可能是阻断SVCV感染的潜在靶点。【结论】成功克隆了鲤RNF14基因的基因编码区,揭示了鲤RNF14基因在SVCV和poly(I:C)刺激下免疫响应规律。【Objective】This study aims to explore the biological function of ring finger protein 14 gene(RNF14)in Cyprinus carpio,and to provide new ideas for blocking spring viremia of Cyprinus carpio virus(SVCV)infection.【Method】The function of RNF14 gene in Cyprinus carpio was preliminarily analyzed by PCR,bioinformatics analysis and real-time fluorescence quantitative PCR,and its expre-ssion pattern after virus stimulation was studied.【Result】RNF14 gene in Cyprinus carpio was 1362 bp in length,encoding 453 amino acids.The RNF14 protein of Cyprinus carpio had closer relationship with Sinocyclocheilus rhinocerous and farther relationship with Columba livia.The protein is an acidic unstable hydrophilic protein;Functional domain prediction showed that the protein was a typical RBR E3s.The results of ubiquitination substrate prediction showed that the protein interacted with RBCK1,Nedd4l,Notch1a and other proteins with higher confidence.Tissues expression profile analysis showed that RNF14 gene was widely expressed in various tissues or organs,with the highest expression in the heart.Compared with the control group,the mRNA expression level of RNF14 gene was significantly increased in muscle at 24 h after high-dose SVCV infection.The mRNA expression level of RNF14 gene was significantly increased in kidney and head kidney after 24 h of Ploy(I:C)stimulation in vivo.Compared with the control group,the mRNA expression level of RNF14 gene in Cyprinus carpio was significantly increased at each sampling time point after low-dose SVCV and Ploy(I:C)infection in vitro.The results of pathogen stimulation in vivo and in vitro showed that the RNF14 gene of Cyprinus carpio was involved in the immune response process after pathogen stimulation,suggesting that the RNF14 gene of Cyprinus carpio may be a potential target for blocking SVCV infection.【Conclusion】The CDS region of RNF14 gene in Cyprinus carpio was successfully cloned,and the immune response of RNF14 gene in Cyprinus carpio under SVCV and Ploy(I:C)stimulation was

关 键 词：鲤 环指蛋白14 生物信息学 鲤春季病毒血症病毒 多肌胞苷酸 病原刺激 免疫响应 

分 类 号：S917.4[农业科学—水产科学]