HE基因受体结合域变异对牛冠状病毒感染的影响  

Effect of HE Gene Receptor Binding Domain Variation on Bovine Coronavirus Infection

作　　者：姜慧华 赵龙 郭抗抗[1] JIANG Huihua;ZHAO Long;GUO Kangkang(College of Veterinary Medicine,Northwest A&F University,Yangling 712100,China)

机构地区：[1]西北农林科技大学动物医学院,杨凌712100

出　　处：《畜牧兽医学报》2025年第3期1336-1343,共8页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：宁夏回族自治区重点研发计划(2021BEF03005);青海省科技计划项目(2022-NK-118);西藏自治区科技计划项目(XZ202101YD0001C)。

摘　　要：为研究HE基因受体结合域变异对牛冠状病毒(BCoV)感染的影响,构建了pEGFP-HE420、pEGFP-HE424和pEGFP-HE428重组质粒。三种重组质粒分别转染至HCT-8细胞,然后感染NX-2021-B2-HE424分离株通过RT-qPCR检测病毒mRNA在吸附、内化和复制阶段的表达水平以及通过TCID 50检测胞内和胞外的病毒滴度。结果显示,pEGFP-HE420、pEGFP-HE424和pEGFP-HE428三种重组质粒均能在HCT-8细胞中表达。与pEGFP-HE424相比,HE基因受体结合域缺失或插入不影响病毒的吸附、内化、复制及胞内病毒滴度;HE基因受体结合域缺失降低了BCoV胞外病毒滴度;HE基因受体结合域插入不影响BCoV胞外病毒滴度。本研究通过构建仅受体结合域变异的三种HE基因的重组质粒在HCT-8细胞上探究了其对牛冠状病毒感染的影响,为研究牛冠状病毒的感染机制提供了数据支撑。To study the effect of HE gene receptor binding domain variation on bovine coronavirus(BCoV)infection,recombinant plasmids pEGFP-HE420,pEGFP-HE424 and pEGFP-HE428 were constructed.Three recombinant plasmids were transfected into HCT-8 cells respectively and then infected with NX-2021-B2-HE424 to detect viral mRNA expression levels at adsorption,internalization and replication stages by RT-qPCR,and to detect viral intracellular and extracellular titers by TCID 50.The results showed that pEGFP-HE420,pEGFP-HE424 and pEGFP-HE428 could be expressed in HCT-8 cells.Compared with pEGFP-HE424,deletion or insertion of HE gene receptor binding domain did not affect the adsorption,internalization,replication and intracellular virus titer.Deletion of HE gene receptor binding domain decreased extracellular titers of BCoV.HE gene receptor binding domain insertion did not affect the BCoV extracellular virus titer.In this study,three recombinant plasmids of HE genes with only receptor binding domain variation was constructed to explore its effect on bovine coronavirus infection in HCT-8 cells,providing data support for the study of the infection mechanism of BCoV.

关 键 词：牛冠状病毒 HE基因 受体结合域变异 病毒感染 

分 类 号：S852.659.6[农业科学—基础兽医学]