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.
作者
叶晨倩
李智丽
叶曼青
秦文珍
杨心雨
翟雪滢
郑浩
童武
李国新
于海
童光志
单同领
娄华
孔宁
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)
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)。
关键词
猪德尔塔冠状病毒
N蛋白
原核表达
多克隆抗体
Porcine deltacoronavirus
N protein
prokaryotic expression
polyclonal antibody
作者简介
叶晨倩,女,硕士研究生,兽医专业;通信作者:孔宁,E-mail:kongning@shvri.ac.cn;娄华,E-mail:13929960788@163.com。
