表达鸡传染性贫血病毒VP1蛋白的新型重组血清4型禽腺病毒的构建  

Construction of a Novel Recombinant Serotype 4 Fowl Adenovirus Expressing VP1 Protein of Chicken Infectious Anaemia Virus

作　　者：徐贞琦 蒋蕙如 郭艺文 谢泉[1,2,3,4] 殷楠 李拓凡 万志敏 邵红霞 秦爱建[1,2,3,4] 张俊 叶建强[1,2,3,4] XU Zhenqi;JIANG Huiru;GUO Yiwen;XIE Quan;YIN Nan;LI Tuofan;WAN Zhimin;SHAO Hongxia;QIN Aijian;ZHANG Jun;YE Jianqiang(Key Laboratory of Jiangsu Preventive Veterinary Medicine,Key Laboratory for Avian Preventive Medicine Ministry of Education,Yangzhou University,Yangzhou,Jiangsu 225009;Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses,Yangzhou,Jiangsu 225009;Institutes of Agricultural Science and Technology Development,Yangzhou University,Yangzhou,Jiangsu 225009;Joint International Research Laboratory of Agriculture and Agri-Product Safety,Ministry of Education,Yangzhou University,Yangzhou,Jiangsu 225009)

机构地区：[1]扬州大学兽医学院,禽类预防医学教育部重点实验室,江苏省动物预防医学重点实验室,江苏扬州225009 [2]江苏高校动物重要疫病与人兽共患病防控协同创新中心,江苏扬州225009 [3]扬州大学农业科技发展研究院,江苏扬州225009 [4]教育部农业与农产品安全国际合作联合实验室,江苏扬州225009

出　　处：《中国家禽》2024年第4期47-53,共7页China Poultry

基　　金：大学生创新创业训练计划项目(202111117014);江苏高校优势学科建设工程资助项目。

摘　　要：为研制新型鸡传染性贫血(CIA)高效疫苗以解决国内商品鸡群中普遍存在的鸡传染性贫血病毒(CIAV)感染问题,试验利用CRISPR-Cas9技术和Cre-LoxP系统,以血清4型禽腺病毒(FAdV-4)为载体,构建表达CIAV VP1蛋白的重组FAdV-4。通过间接免疫荧光试验和蛋白免疫印迹试验对重组病毒进行验证,通过体外病毒生长曲线来研究其复制效率。结果显示:该重组病毒构建成功,能有效表达CIAV VP1蛋白,命名为rFAdV-4-CIAV-VP1,且发现rFAdV-4-CIAV-VP1在LMH细胞中的复制效率远低于野生型FAdV-4。研究表明构建的重组病毒rFAdV-4-CIAV-VP1为CIAV和FAdV-4的联防联控提供了二联候选疫苗株。To develop a novel highly effective vaccine against chicken infectious anaemia(CIA)to solve the universal infec⁃tion of chicken infectious anaemia virus(CIAV)in commercial chickens in China,a recombinant serotype 4 fowl adenovirus(FAdV-4)expressing VP1 protein of CIAV,was constructed by using FAdV-4 as vector through CRISPR-Cas9 and Cre-LoxP system in this study.The recombinant virus was verified by indirect immunofluorescence assay(IFA)and Western blot(WB),and the replication efficiency of the recombinant virus was studied by viral growth curves in vitro.The results showed that the recombi⁃nant virus was successfully constructed and could effectively express VP1 protein of CIAV,designated as rFAdV-4-CIAV-VP1.The viral growth curve showed that the replication efficiency of rFAdV-4-CIAV-VP1 in LMH cells was much lower than that of wild-type FAdV-4.The results indicated that the construction of the recombinant virus rFAdV-4-CIAV-VP1 expressing VP1 protein of CIAV provided a dual vaccine candidate for the joint prevention and control of CIAV and FAdV-4.

关 键 词：鸡传染性贫血病毒 VP1蛋白 CRISPR-Cas9 重组血清4型禽腺病毒 表达 

分 类 号：S855.3[农业科学—临床兽医学]