虾肝肠胞虫Septin的生物信息学分析及其在增殖中的潜在作用  

Bioinformatics Analysis of Enterocytozoon hepatopenaei Septin and Its Potential Role in Proliferation

作　　者：张晟 乔毅[2] 曹晓慧 张雷霆 成婕 吴海龙 沈辉[2] Zhang Sheng;Qiao Yi;Cao Xiaohui;Zhang Leiting;Cheng Jie;Wu Hailong;Shen Hui(School of Marine Science and Fisheries,Jiangsu Ocean University,Lianyungang 222000,China;Jiangsu Marine Fisheries Research Institute,Nantong 226007,China;School of Marine and Engineering,Nanjing Normal University,Nanjing 210023,China)

机构地区：[1]江苏海洋大学海洋科学与水产学院,江苏连云港222000 [2]江苏省海洋水产研究所,江苏南通226007 [3]南京师范大学海洋科学与工程学院,江苏南京210023

出　　处：《南京师大学报(自然科学版)》2025年第1期66-74,共9页Journal of Nanjing Normal University(Natural Science Edition)

基　　金：江苏省“333”人才专项[(2022)]21;江苏海洋生物资源创新中心科研项目计划专项资金(2022YHTZZZ04);江苏省渔业专项(2023-SJ-072)。

摘　　要：虾肝肠胞虫(Enterocytozoon hepatopenaei,EHP)是对虾养殖业的重要病原体之一,其裂殖增殖的分子机制尚不明确.本研究通过克隆和分析EHP裂殖期Septin蛋白EhpSeptin的基因序列、表达及定位,初步阐释了该基因在EHP裂殖中的潜在作用.结果表明,EhpSeptin基因序列编码265个氨基酸的蛋白质,分子量约30.4 kDa,等电点4.75.构建EhpSeptin原核表达载体,获得高纯度表达蛋白.Western blotting验证了表达蛋白与天然蛋白一致.此外,本研究建立了EhpSeptin基因显色原位杂交技术(chromogenic in situ hybridization,CISH),分析感染对虾样本中EhpSeptin的表达模式.结果显示,EhpSeptin的表达水平随EHP感染强度增加而升高,表明其参与了EHP的裂殖过程.本研究首次报道了EHP裂殖相关蛋白EhpSeptin的克隆、表达、定位及其在EHP感染过程中的变化规律,为探究EHP裂殖分子机制和防治该病原体的感染提供了理论基础和技术手段.Enterocytozoon hepatopenaei(EHP)is one of the significant pathogens in the shrimp aquaculture industry,but the molecular mechanisms of its division and proliferation remain unclear.This study aimed to elucidate the potential role of the EhpSeptin protein during EHP division by cloning and analyzing the gene sequence,expression,and localization of EhpSeptin during the division period of EHP.The results indicated that the EhpSeptin gene sequence encoded a protein with 265 amino acids,a molecular weight of approximately 30.4 kDa,and an isoelectric point of 4.75.An EhpSeptin prokaryotic expression vector was constructed,and high-purity expressed protein was obtained.Western blotting confirmed the consistency of the expressed protein with the native protein.Additionally,this study established chromogenic in situ hybridization(CISH)for the EhpSeptin gene to analyze its expression pattern in infected shrimp samples.The results demonstrated that the expression level of EhpSeptin increased with the intensity of EHP infection,suggesting its involvement in the EHP division process.This study is the first to report the cloning,expression,localization,and variations in the EhpSeptin protein during EHP infection,providing a theoretical foundation and technical means for exploring the molecular mechanisms of EHP division and preventing infection by this pathogen.

关 键 词：虾肝肠胞虫 SEPTIN 原核表达 显色原位杂交 

分 类 号：S945.4[农业科学—水产养殖]