摘要
旨在为解决国内禽白血病净化周期长以及净化鸡群J亚群禽白血病病毒(ALV-J)感染频发等问题提供新型抗ALV-J策略。本研究利用前期构建好的增强型绿色荧光蛋白(EGFP)与血清4型禽腺病毒(FAdV-4)纤突蛋白Fiber-2融合的重组FAdV-4为载体,并使用CRISPR/Cas9和Cre-LoxP技术构建表达ALV-J囊膜蛋白(Env)的重组FAdV-4,通过间接免疫荧光试验和Western blot验证重组病毒表达的Env蛋白,测定重组病毒在鸡肝癌细胞系(LMH)细胞中的增殖特性。结果:成功拯救了能表达ALV-J Env蛋白的重组病毒FAdV-4-ALV-J-env,该重组病毒在LMH细胞中的复制效率远低于野生型FAdV-4。本研究构建的重组病毒FAdV-4-ALV-J-env为ALV-J和FAdV-4感染的联防联控提供了二联候选疫苗株。
A novel anti-ALV-J strategy is urgently needed to help to solve the long eradication time-course and frequent re-emerging of ALV-J infection in eradicated chickens.This study utilized a previously constructed Enhanced green fluorescent protein(EGFP)and partially used Fiber-2 recombinant serotype 4(FAdV-4)avian adenovirus(FAdV-4)as vector,and further constructed a recombinant FAdV-4 expressing ALV-J Env protein using CRISPR/Cas9 and Cre-LoxP techniques.The Env protein expressed by the recombinant virus was detected through indirect immunofluorescence and Western blot,and the proliferation characteristics of the recombinant virus were evaluated in LMH cells.Our results showed that the recombinant virus FAdV-4-ALV-J-env expressing ALV-J Env protein was successfully rescued and the replication efficiency of this recombinant virus in LMH cells was much lower than that of the wild-type FAdV-4.All these indicated that the FAdV-4-ALV-J-env constructed here provided a bivalent vaccine candidate for the prevention and control of ALV-J and FAdV-4.
作者
徐超
谢泉
武佳妍
王胜男
万志敏
邵红霞
秦爱建
叶建强
李拓凡
XU Chao;XIE Quan;WU Jiayan;WANG Shengnan;WAN Zhimin;SHAO Hongxia;QIN Aijian;YE Jianqiang;LI Tuofan(College of Veterinary Medicine Yangzhou University/inistry of Education Key Laboratory for Avian Preventive Medicine/Key Laboratory of Jiangsu Preventive Veterinary Medicine,Yangzhou 225009,China;Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses,Yangzhou 225009,China)
出处
《畜牧与兽医》
北大核心
2025年第3期66-73,共8页
Animal Husbandry & Veterinary Medicine
基金
国家自然科学基金青年项目(32302839)
江苏省自然科学基金青年项目(BK20220579)
江苏省农业科技自主创新资金项目(CX(23)3075)。
