文献类型：期刊文献

中文题名：尼罗罗非鱼LCP1基因的克隆、表达及功能分析

英文题名：Molecular cloning of lymphocyte cytosolic protein 1(LCP1)gene in Nile Tilapia(Oreochromis niloticus)and its role in the immune response against Streptococcus agalactiae

作者：李伟富[1,2];袁嘉翊[1,2];张湘东[1,2];汪志文[1,2];夏洪丽[1,2];鲁义善[1,2]

机构：[1]广东海洋大学水产学院,广东省水产动物病害防控与健康养殖重实验室,广东湛江524088;[2]广东海洋大学深圳研究院,广东省水生动物健康评估工程技术研究中心,深圳市海水经济动物种苗健康评价公共技术服务平台,广东深圳518120

年份：2025

卷号：40

期号：5

起止页码：761

中文期刊名：大连海洋大学学报

外文期刊名：Journal of Dalian Ocean University

收录：北大核心2023、、北大核心

基金：广东省基础与应用基础研究基金(2021A1515110214)。

语种：中文

中文关键词：尼罗罗非鱼;LCP1基因;无乳链球菌;mRNA表达分析

外文关键词：Oreochromis niloticus;lymphocyte cytosolic protein(LCP1)gene;Streptococcus agalactiae;mRNA expression analysis

中文摘要：为研究尼罗罗非鱼(Oreochromis niloticus)LCP1基因的分子特征及其在抵抗无乳链球菌免疫应答中的潜在功能,从尼罗罗非鱼中克隆出了LCP1基因,命名为OnLCP1,并对该基因进行生物信息学分析和亚细胞定位分析;运用荧光定量PCR技术分析了经灭活无乳链球菌免疫刺激后OnLCP1在各组织中mRNA的表达情况;采用双荧光素酶报告系统检测了过表达LCP1对NF-κB、STAT1、ISRE、IFN-1和IFN-3信号通路活性的影响。结果显示,OnLCP1包含1857个碱基对,可编码一个618氨基酸的蛋白质,On LCP1蛋白被鉴定含有EF臂结构域(钙结合结构域)和两个串联的肌动蛋白结合结构域(ABD),每个肌动蛋白结合结构域均包含两个钙调蛋白同源(CH)结构域;On LCP1是一个无跨膜的细胞外蛋白,与人类LCP1蛋白的三维结构高度相似,表明其功能可能是保守的;荧光定量PCR分析显示,OnLCP1基因在健康尼罗罗非鱼的不同组织中均有表达,尤其在脾脏中表达量最高;研究发现,经灭活无乳链球菌免疫后,OnLCP1基因在免疫器官中的表达量显著增加,表明其可能参与了对病原菌的免疫应答;亚细胞定位显示,OnLCP1在HEK-293T细胞的细胞质中表达;双荧光素酶报告系统显示,OnLCP1显著提高了NF-κB信号通路的活性(P<0.05),表明该分子可能通过NF-κB通路介导罗非鱼抵御无乳链球菌感染。本研究结果为进一步研究LCP1基因在鱼类先天免疫中的功能提供了科学参考。

外文摘要：To investigate molecular characteristics of the Nile tilapia(Oreochromis niloticus)lymphocyte cytosolic protein(LCP1)gene and its potential function in the immune response against Streptococcus agalactiae,the gene encoding LCP1 from Nile tilapia was successfully cloned(OnLCP1).Bioinformatics analysis was conducted on this gene,and its subcellular localization was determined.The expression pattern of OnLCP1 mRNA in various tissues after S.agalactiae stimulation was analyzed by using fluorescence quantitative PCR.A dual luciferase reporter gene assay was used to detect the impact of transcription factors such as NF-κβ,STAT1,ISRE,IFN-1,and IFN-3 on promoter activity of the LCP1 gene.The results showed that OnLCP1 consists of 1857 base pairs and encodes a 618-amino acid protein.The OnLCP1 protein contains an EF-hand domain(calcium binding domain)and two tandem actin binding domains(ABD),with each actin domain containing two calmodulin homology(CH)domains.OnLCP1 is an extracellular protein without the transmembrane region,and it shares a highly similar three-dimensional structure with the human LCP1 protein,suggesting that its function may be conserved.OnLCP1 was expressed in various tissues of healthy Nile tilapia,with the highest expression level in the spleen.After stimulation by inactivated S.agalactiae,the expression of the OnLCP1 gene in immune organs significantly increased,indicating its potential involvement in the immune response against pathogenic bacteria.The subcellular localization experiment showed that OnLCP1 was expressed in the cytoplasm of HEK-293T cells.The dual luciferase reporter gene experiment showed that the OnLCP1 gene significantly activates the NF-κβsignaling pathway(P<0.05),suggesting that OnLCP1 may mediate defense against S.agalactiae infection through this pathway.The results of this study provide a scientific ground for further research of LCP1 function in the innate immunity of fish.