血清4型禽腺病毒纤突蛋白的真核表达及其免疫反应性  

Eukaryotic Expression of Fiber Gene of Serotype 4 Fowl Adenovirus and Their Immunoreactivity

作　　者：王伟康 梁广成[1,2] 管雪冰 张俊 王萍 张建军 郑文铝 邵红霞 秦爱建[1,2] 叶建强 WANG Weikang;LIANG Guangcheng;GUAN Xuebing;ZHANG Jun;WANG Ping;ZHANG Jianjun;ZHENG Wenlu;SHAO Hongxia;QIN Aijian;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;Sinopharm Yangzhou VAC Biological Engineering Co., Ltd, Yangzhou, Jiangsu 225127;Joint International Research Laboratory of Agriculture and Agri-Product Safety, the Ministry of Education of China, 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]国药集团扬州威克生物工程有限公司,江苏扬州225127 [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》2017年第23期14-17,共4页China Poultry

基　　金：江苏高校优势学科建设工程资助项目;江苏省双创团队项目;扬州大学大学生科技创新项目(X20160704)

摘　　要：研究对血清4型禽腺病毒(FAdV-4)表面纤突蛋白基因F1及F2进行了克隆及真核表达载体的构建,分别命名为pcDNA3.1-F1以及pcDNA3.1-F2。间接免疫荧光以及Western blot分析证明,所构建的表达载体表达的F1及F2均能与抗FAdV-4的阳性血清进行良好免疫反应。在间接免疫荧光试验中,转染pcDNA3.1-F1及pcDNA3.1-F2的293T细胞内均可见特异性亮绿色荧光;在Western blot中分别出现F_1及F_2特异性蛋白条带。纤突蛋白F1及F2真核表达载体构建、表达及其良好的免疫反应性,为进一步建立FAdV-4快速血清学诊断方法、探究纤突蛋白F_1及F_2在病毒感染致病中作用奠定了基础。In this study, fiberl and fiber2 genes of serotype 4 fowl adenovirus （FAdV-4）were cloned, and their eukaryotic expression vectors were constructed, designated pcDNA3.1-F1 and pcDNA3.1-F2 respectively. Indirect immunofluoreseent assay （IFA）and western blot analysis revealed that the expression product of F1 and F2 could react well with sera against FAdV-4. In IFA, the specific bright fluorescence could be found in the cells transfected with pcDNA3.1-F1 and peDNA3.1-F2 respectively. And a specific band of Fiberl and Fiber2 protein could be found in cells transfected with pcDNA3.1-F1 and pcDNA3.1-F2 respectively in western blot analysis. Overall, the construction and expression of fiberl and fiber2 of FAdV-4, and their good immunoreactivity pave the ways for further establishing a serological diagnostic method for FAdV-4 and exploring the roles of Fibers in the infection and pathogensis of FAdV-4.

关 键 词：血清4型禽腺病毒 纤突蛋白基因 克隆表达 免疫反应性 

分 类 号：S855.3[农业科学—临床兽医学]