猪德尔塔冠状病毒E蛋白真核表达及遗传进化分析  

PDCoV E Protein prokaryotic expression and phylogenetic analysis

作　　者：江乐 高佳龙 玉山·艾合麦特 乃斯如拉·麦麦提明 妥小丽 弓超 JIANG Le;GAO Jialong;YU Shan·Aihemaite;NAI Sirula·Maimaitiming;TUO Xiaoli;GONG Chao(College of Veterinary Medicine,Xinjiang Agricultural University,Urumqi 830052,China)

机构地区：[1]新疆农业大学,新疆乌鲁木齐830052

出　　处：《现代牧业》2024年第3期17-22,共6页Modern Animal Husbandry

基　　金：新疆农业大学校级大学生创新项目(DXSCX2023215)。

摘　　要：对猪德尔塔冠状病毒毒株的E蛋白进行真核表达并进行系统进化分析,为下一步探索E基因在PDCoV的生物学特性和诊断方法的建立奠定试验基础。本研究以PDCoV-HNZMD的RNA为模板,扩增PDCoV E基因,将其插入真核表达载体pcDNA3.1(+)并构建真核表达重组质粒pcDNA3.1-E。之后转染HEK-293T细胞,采用IFA和western blot法检测基因的表达结果。成功获得了PDCoV E蛋白目的基因,并成功构建真核载体pcDNA3.1-E。IFA和Western blot检测结果表明,构建的重组真核表达质粒正确表达氨基酸同源性的比例为92.9%~100%,遗传进化树分析表明,PDCoV-HNZMD与中国广西毒株(OQ736717)亲缘关系最为相近。成功构建了PDCoV-E蛋白的真核表达质粒,并进行了IFA和WB的验证,为后续致病机制的研究提供参考。The eukaryotic expression of E protein in Porcine Delta Coronavirus strain was analyzed and phylogenetic analysis was performed,which laid a foundation for the further exploration of the biological characteristics of E gene in PDCoV and the establishment of diagnostic methods.In this study,PDCoV E gene was amplified by reverse transcription using PDCoV-HNZMD RNA as template,ligated and transformed to construct the eukaryotic expression recombinant plasmid pcDNA3.1-E.After transfected with HEK-293T cells,the gene expression was detected by IFA and western blot.The target gene of PDCoV E protein was successfully obtained,and the eukaryotic vector pcDNA3.1-E was successfully constructed.The results of IFA and Western blot showed that the recombinant eukaryotic expression plasmid could correctly express,the proportion of amino acid homology was 92.9%-100%.The genetic evolution tree analysis showed that PDCOV-HNZMD was most closely related to the Guangxi strain(OQ736717).In this study,the eukaryotic expression plasmid of PDCoV-E protein was successfully constructed and verified by IFA and WB,which provides a reference for the subsequent research on the pathogenic mechanism.

关 键 词：猪德尔塔冠状病毒 E蛋白 真核表达 遗传进化分析 

分 类 号：S482.3[农业科学—农药学]