罗非鱼多聚免疫球蛋白受体的生物学特性及对罗非鱼湖病毒增殖的影响  

Biological characteristics of tilapia pIgR and its effect on the proliferation of tilapia lake virus

作　　者：李波 郑树城 曾伟伟 王庆[2] 李莹莹[2] 尹纪元 王雅慧 吕月凤 石存斌[2] 赵志英 姜志勇 王英英[2] LI Bo;ZHENG Shu-cheng;ZENG Wei-wei;WANG Qing;LI Ying-ying;YIN Ji-yuan;WANG Ya-hui;LV Yue-feng;SHI Cun-bin;ZHAO Zhi-ying;JIANG Zhi-yong;WANG Ying-ying(College of Fisheries and Life Sciences,Shanghai Ocean University,Shanghai 201306,China;Pearl River Fishery Research Institute,Chinese Academy of Fishery Sciences/Key Laboratory of Fishery Drug Development,Ministry of Agriculture and Rural Affairs,Guangdong Key Laboratory of Aquatic Animal Immune Technology,Guangzhou 510380,China;College of Life Science and Engineering,Key Laboratory of Animal Molecular Design and Precision Breeding,Foshan University,Foshan 440605,Guangdong,China;College of Ocean and Earth Sciences,Xiamen University,Xiamen 361100,Fujian,China;Hainan Academy of Ocean and Fisheries Sciences,Haikou 571126,China;Guangdong Provincal Agricultural Technology Extension Center,Guangzhou 510030,China)

机构地区：[1]上海海洋大学水产与生命科学学院,上海201306 [2]中国水产科学研究院珠江水产研究所,农业农村部渔用药物创制重点实验室,广东省水产动物免疫技术重点实验室,广州510385 [3]佛山科学技术学院,动物分子设计与精准育种重点实验室,普通高校动物分子设计与精准育种重点实验室,生命科学与工程学院,广东佛山440605 [4]厦门大学海洋与地球学院,福建厦门361100 [5]海南省海洋与渔业科学院,海口571126 [6]广东省农业技术推广中心,广州510030

出　　处：《淡水渔业》2022年第4期85-91,共7页Freshwater Fisheries

基　　金：国家重点研发计划(2019YFD0900101);广东省现代农业产业技术体系创新团队建设专项(2021KJ150;2021KJ119);国家大宗淡水鱼产业技术体系(CARS-45);海南省本级部门预算项目(2018)。

摘　　要：罗非鱼湖病毒TiLV是一种新发RNA病毒,对罗非鱼养殖业造成巨大威胁。为明确罗非鱼(Oreochromis niloticus)多聚免疫球蛋白受体基因(OnpIgR)的序列和结构特征及其对TiLV增殖的影响,本研究首先对PCR扩增获得的OnpIgR完整的ORF,进行了结构域预测、三维结构预测以及遗传进化树分析,然后采用荧光定量PCR方法分析了罗非鱼感染TiLV后该基因在各组织的表达模式,最后构建了OnpIgR过表达质粒,初步研究了OnpIgR对TiLV增殖的影响。结果表明,PCR扩增获得了完整的OnpIgR ORF序列,长度为1023 bp,该ORF序列具有一个信号肽,两个Ig样结构域,同源建模结果显示,pIgR蛋白整体呈扭曲的“L”型,主要由反平行β片层和无规卷曲组成;系统发育树显示OnpIgR与大口黑鲈(Micropterus salmoides)以及斜带石斑鱼(Epinephelu coioides)的pIgR聚为一支;感染TiLV后,罗非鱼皮肤、脾脏以及肾脏中pIgR均呈现先上升后下降的趋势;成功构建了pEGFP-OnpIgR并在TiB细胞中过表达OnpIgR后感染TiLV,发现TiLV的S8和S10节段mRNA相对表达量显著增多,TiLV拷贝数也显著增多,初步表明OnpIgR对TiLV的增殖具有促进作用。Polymeric immunoglobulin receptor(pIgR)plays an essential role in pathogen infection and invasion.Tilapia lake virus(TiLV)was a novel RNA virus that harms the tilapia and its hybrids and cause a great threat to Tilapia culture industry.Our previous study showed that pIgR expression levels in tilapia tissues changed significantly after TiLV infection.This experiment was conducted to understand the biological characteristics of tilapia(Oreochromis niloticus)pIgR(OnpIgR)and its effects on TiLV infection and proliferation.Firstly,the complete open reading frame of OnpIgR was amplified,and its structure domain prediction,three-dimensional structure prediction and phylogenetic tree construction were carried out.Secondly,the pIgR expression level in spleen,kidney and skin tissues of tilapia infected with TiLV was analyzed by quantitative fluorescence.Finally,the recombinant eukaryotic expression vector pEGFP-OnpIgR was constructed and transfected into Tilapia brain cell(TiB)to study overexpression OnpIgR on TiLV proliferation.The results showed that the complete OnpIgR ORF sequence was obtained by PCR amplification and the full length of ORF was 1023bp.Tilapia pIgR was consisted of a signal peptide,two Ig-like domains and a transmembrane domain.The phylogenetic tree showed that pIgR was clustered with the pIgR of Micropterus salmoides and Epinephelus coioides.Fluorescence quantitative PCR analysis showed that pIgR increased first and then decreased in skin,spleen and kidney tissues of tilapia.TiB cells were infected with TiLV after overexpressing OnpIgR,and it was found that the relative expression levels of RNA in S8 and S10 segments of TiLV were significantly increased,as well as the copy number of TiLV genome,which preliminarily indicated that OnpIgR could promote the proliferation of TiLV.In conclusion,results showed that TiLV might stimulate the expression of OnpIgR and the overexpression of pIgR in TiB cell can promote the proliferation of TiLV.This study may provide an important reference for revealing the mechan

关 键 词：罗非鱼湖病毒 黏膜免疫 多聚免疫球蛋白受体 过表达 

分 类 号：S917.4[农业科学—水产科学]