摘要
研究首先对血清8型禽腺病毒(FAd V-8)的纤突蛋白基因F进行了克隆并分别构建了原核表达载体和真核表达载体,经测序验证后分别命名为p GEX-6P-1-F和pc DNA3.1-F。将原核表达质粒pGEX-6P-1-F转化BL21,经IPTG诱导,获得了FAd V-8 F蛋白的GST-F可溶性融合表达产物。将纯化的GST-F蛋白免疫BALB/c小鼠制备了抗FAd V-8 F蛋白的多克隆抗体。Western blot结果证明,制备的抗FAd V-8 F蛋白的多克隆抗体能特异性地识别转染pc DNA3.1-F或感染FAd V-8病毒细胞中57 ku大小的蛋白条带。间接免疫荧光试验同样证明,抗FAd V-8的F蛋白的多克隆抗体具有良好的反应性及特异性。研究结果为进一步建立FAd V-8快速血清学诊断方法、亚单位疫苗研制及探究纤突蛋白F在病毒感染致病中作用奠定了基础。
In this study, fiber gene of serotype 8 fowl adenovirus （FAdV-8）was cloned, and inserted into prokaryotic expression vector and eukaryotic expression vector, designated pGEX-6p-1-F and pcDNA3.1-F respectively. The pGEX-6p-1-F was transformed into E. coli BL21 and induced by IPTG to obtain the recombinant fused protein GST-F. The polyclonal antibodies against the fiber protein of FAdV-8 were generated through immunizing BALB/c mice with the purified GST-F protein. Western blot showed that the polyclonal antibodies against F protein of FAdV-8 could specifically recognize 57 ku protein bands in the cells transfected with pcDNA3.1-F or infected with FAdV-8 respectively. Indirect immunofluorescence assay also confirmed the reactivity and specificity of polyclonal antibodies against F protein of FAdV-8. All these paved the ways for further establishing rapid serological diagnostics tools, developing subunit vaccine for FAdV-8, and exploring the role of the fiber in the infection and pathogenesis of FAdV-8.
作者
路浩
张伟
王伟康
梁广成
王萍
张建军
郑文铝
邵红霞
秦爱建
邹海涛
叶建强
LU Hao;ZHANG Wei;WANG Weikang;LIANG Guangcheng;WANG Ping;ZHANG Jianjun;ZHENG Wenlv;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, Ministry of Education, Yangzhou University, Yangzhou, Jiangsu 225009)
出处
《中国家禽》
北大核心
2018年第4期16-20,共5页
China Poultry
基金
江苏高校优势学科建设工程资助项目
关键词
血清8型禽腺病毒
纤突蛋白
克隆表达
多克隆抗体
Serotype 8 fowl adenovirus
fiber gene
cloning and expression
polyclonal antibodies
