中国畜牧兽医2016，Vol.43Issue(8)：1967-1974,8.DOI:10.16431/j.cnki.1671-7236.2016.08.006
鸡传染性支气管炎病毒 HH06株囊膜蛋白多克隆抗体制备及生物学功能检测
Preparation of Polyclonal Antibody against Recombinant E Protein of IBV HH06 Strain and Its Biological Function
摘要
Abstract
In this study ,IBV HH06 complete E gene was firstly cloned and sequenced .According to the hydrophilicity and antigenic index analysis ,its partial gene (193 to 327 bp) was subcloned into prokaryotic expression vector pET‐32a(+ ) and eukaryotic expression vector PVAX1 .The recombinant plasmid pET‐32a‐E1 was transformed into E .coli Rosetta (DE3) and induced with IPTG .The recombinant IBV truncated E1 protein with molecular weight of 23 ku was observed as expected .It could be recognized by positive IBV antisera in Western blotting with high reactivity . Then the purified recombinant protein was used as antigen for immunization of rabbit to prepare polyclonal antibody .Indirect ELISA showed that the titer of polyclonal antibody was 220 ,and it had high reactivity and specialty with recombinant protein .Furthermore ,IFA test demonstrated that this polyclonal antibody could react with Hela cells transfected with PVAX‐E1 plasmid and IBV‐infected CEK cells .T he IBV E polyclonal antibody obtained in this study laid a foundation for further functional research of E protein in IBV pathogenesis .关键词
传染性支气管病毒/囊膜蛋白/多克隆抗体/免疫印迹试验/间接免疫荧光试验Key words
IBV/envelope protein/polyclonal antibody/Western blotting/indirect fluorescence an-tibody assay分类
农业科技引用本文复制引用
白妍,赵蕾,杜威威,潘龙,黄小丹,杨贵君,李广兴..鸡传染性支气管炎病毒 HH06株囊膜蛋白多克隆抗体制备及生物学功能检测[J].中国畜牧兽医,2016,43(8):1967-1974,8.基金项目
国家自然科学基金项目 （）
