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.
作者
徐贞琦
蒋蕙如
郭艺文
谢泉
殷楠
李拓凡
万志敏
邵红霞
秦爱建
张俊
叶建强
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)
出处
《中国家禽》
北大核心
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。
