猪德尔塔冠状病毒N蛋白原核表达及多克隆抗体制备  

Prokaryotic Expression of N Protein of Porcine Deltacoronavirus and Preparation of Polyclonal Antibodies

作　　者：叶晨倩 李智丽 叶曼青 秦文珍 杨心雨 翟雪滢 郑浩[2] 童武[2] 李国新[2] 于海[2] 童光志[2] 单同领[2] 娄华 孔宁[2] YE Chenqian;LI Zhili;YE Manqing;QIN Wenzhen;YANG Xinyu;ZHAI Xueying;ZHENGHao;TONG Wu;LI Guoxin;YU Hai;TONG Guangzhi;SHAN Tongling;LOU Hua;KONG Ning(Foshan University,Foshan 528000,China;Shanghai Veterinary Research Institute,CAAS,Shanghai 200241,China)

机构地区：[1]佛山大学,佛山528000 [2]中国农业科学院上海兽医研究所,上海200241

出　　处：《中国动物传染病学报》2025年第1期164-170,共7页Chinese Journal of Animal Infectious Diseases

基　　金：国家重点研发项目(2021YFD1801102);中央级公益性科研院所基本科研业务费专项(Y2022QC28)。

摘　　要：猪德尔塔冠状病毒(PDCoV)是一种新发的病原体,引起仔猪腹泻、脱水和呕吐等症状。本研究以PDCoV的RNA为模板,扩增PDCoV N基因,将其插入原核表达载体pColdⅠ中,构建PDCoV N-pColdⅠ质粒;通过BL21(DE3)原核表达和纯化获得高纯度的PDCoV N蛋白;免疫BALB/c小鼠,制备PDCoV N蛋白多克隆抗体。结果显示:原核表达的PDCoV N蛋白为可溶性蛋白,分子量约为40 kDa;通过ELISA、Western blot和IFA鉴定可知,本研究制备的PDCoV N多克隆抗体均具有良好的效价,可为深入开展PDCoV相关的基础和应用研究提供工具。Porcine deltacoronavirus(PDCoV)is an emerging pathogen that causes diarrhea,dehydration and vomiting in pigs.In this study,the RNA of PDCoV was used as a template to amplify its N gene,which was then inserted into the prokaryotic expression vector pColdⅠ.The PDCoV N-pColdⅠplasmid was constructed for transformation into the competent BL21(DE3)cells.The PDCoV N protein was obtained from the prokaryotic expression and purified.The purified PDCoV N protein was used to immunize BALB/c mice for preparation of polyclonal antibodies.The results showed that the PDCoV N protein was in the soluble form with molecular weight of about 40 kDa.The PDCoV N polyclonal antibodies prepared in this study had good reactivity in ELISA,Western blot and IFA and provided tools for in-depth basic and applied research related to PDCoV.

关 键 词：猪德尔塔冠状病毒 N蛋白 原核表达 多克隆抗体 

分 类 号：S852.651[农业科学—基础兽医学]