脊尾白虾Duox基因鉴定及其免疫功能研究  

Identification of the Duox gene in Exopalaemon carinicauda and preliminary study on its immune function

作　　者：刘德雪 范梦昊 程伟涛 曹宴榕 韩婉钰 胡广伟 LIU Dexue;FAN Menghao;CHENG Weitao;CAO Yanrong;HAN Wanyu;HU Guangwei(Jiangsu Key Laboratory of Marine Bioresources and Environment,Jiangsu Key Laboratory of Marine Biotechnology School of Marine Science and Fisheries,Jiangsu Ocean University,Lianyungang 222005,China;Co-Innovation Center of Jiangsu Marine Bio-Industry Technology,Jiangsu Ocean University,Lianyungang 222005,China)

机构地区：[1]江苏海洋大学江苏省海洋生物资源与环境重点实验室,江苏省海洋生物技术重点实验室,江苏连云港222005 [2]江苏海洋大学江苏省海洋生物产业技术协同创新中心,江苏连云港222005

出　　处：《大连海洋大学学报》2025年第1期34-43,共10页Journal of Dalian Ocean University

基　　金：江苏高校“青蓝工程”资助项目;连云港市重点研发项目(SF2303);连云港市“521高层次人才培养工程”(2021-1021);江苏省大学生创新创业训练项目(202311641038Z)。

摘　　要：为了探究Duox基因在脊尾白虾(Exopalaemon carinicauda)中的免疫功能,从脊尾白虾(平均体质量为1.39 g±0.39 g)中成功克隆获得Duox基因cDNA序列,并对其编码的氨基酸序列进行了分析;在此基础上采用实时荧光定量PCR技术(qRT-PCR),分析了该基因的组织表达特征及其在副溶血弧菌(Vibrio parahaemolyticus)感染后的表达变化情况;最后基于RNA干扰技术分析了脊尾白虾Duox基因敲降后感染副溶血弧菌的死亡率。结果表明:脊尾白虾Duox基因编码区序列长度为4518 bp,编码1505个氨基酸,具有双氧化物酶典型结构特征;Duox基因在脊尾白虾不同组织中均有表达,且在鳃组织中表达最高;副溶血弧菌感染后,Duox基因在鳃组织中的表达量显著上升(P<0.05);注射siRNA试剂后用副溶血弧菌感染,与对照组相比,干扰组脊尾白虾的存活率显著降低(P<0.05)。研究表明,Duox基因在脊尾白虾抵御病原菌感染的免疫反应中发挥作用,该研究结果为进一步探究Duox在脊尾白虾中的免疫功能和作用机制提供了有益参考。Dual oxidase(Duox)is a member of the nicotinamide adenine dinucleotide(NADPH)oxidase(NOX)family,which plays a crucial role in various biological processes,such as the production of reactive oxygen species,regulation of the biological clock,and the immune response.To explore the immune function of the Duox gene in Exopalaemon carinicauda,the cDNA sequence of the Duox gene was successfully cloned from E.carinicauda,and the amino acid sequence which it encoded was analyzed.Real-time fluorescence quantitative PCR technology(qRT-PCR)was used to analyze the tissue expression characteristics of gene and its expression changes infected with Vibrio parahaemolyticus.Finally,the mortality rate of E.carinicauda infected with V.parahaemolyticus after Duox gene knockdown was analyzed by using the RNA interference technique.The result shows that the coding region sequence length of the Duox gene in E.carinicauda is 4518 bp,encoding 1505 amino acids,reflecting typical structural characteristics of the dual oxidase.The Duox gene is expressed in all of the E.carinicauda tissues tested,with the highest expression level in gill tissues.After infected with V.parahaemolyticus,the Duox gene expression level in gills increases significantly(P<0.05).Infection with V.parahaemolyticus after injection of siRNA reagent significantly reduces the survival rate of E.carinicauda in the RNA interference group compared with the control group(P<0.05).This study shows that the Duox gene plays a role in the immune response of E.carinicauda against pathogenic infection,and provides an favorable reference for further exploration into the immune function and mechanism of Duox in E.carinicauda.

关 键 词：脊尾白虾 Duox基因 免疫功能 RNAI 

分 类 号：S917.4[农业科学—水产科学]