副鸡禽杆菌抗原表位的筛选与鉴定  

Screening and identification of an epitope of Avibacterium paragallinarum from a phage display peptide library

作　　者：王宏俊[1] 龚玉梅[1] 高亚萍[2] 柳川[2] 张培君[1] 

机构地区：[1]北京市农林科学院畜牧兽医研究所,北京100097 [2]军事医学科学院基础研究所,北京100850

出　　处：《中国兽医科学》2009年第7期587-592,共6页Chinese Veterinary Science

基　　金：国家高技术研究发展计划(863)项目(2005AA246050);北京市科技新星计划项目(2005B35)

摘　　要：用C型副鸡禽杆菌(Apg)单克隆抗体作为靶标,筛选噬菌体展示随机12肽库,从中获得与之特异性结合的噬菌体克隆,建立了一种筛选副鸡禽杆菌表位的方法。测序结果显示,第3轮筛选后,80%的噬菌体克隆具有氨基酸共同序列Y-P-Q(A)WW,含有上述共有序列的噬菌体克隆不仅可以与靶抗体特异结合,还可与C型Apg细菌竞争结合靶抗体;将编码YSPHQWWLSGAV的DNA片段插入质粒pFliTrx的多克隆位点,转化E.coliGI826并在鞭毛成功展示;该重组菌能与靶抗体特异结合;用重组菌免疫试验鸡能产生C型Apg细菌的特异性抗体;用C型668株Apg攻毒,免疫接种鸡获得37.5%保护。提示该表位肽用于研制鸡传染性鼻炎疫苗具有广阔的应用前景。Using the monoclonal antibody（McAb） directed to Avibacterium paragallinarum serotype C as the target protein,phage clones binding to the target protein were screened in the 12-mer random peptide library. More than 80% of the phage clones selected in the third round carried the consensus peptide motif sequence Y-P-Q（A）WW. The phage clones with the peptide motif could react with the target MeAb, which could be competitively inhibited by A. paragallinarum. One of the peptide sequences, YSPHQW WLSGAV,which was itself expressed with the flagella on the Escherichia coli GI826 cell surface was se lected and inserted into the multiple cloning sites of pFliTrx vector. Intramuscular injection of the epitopecontaining bacteria into chickens resulted in production of the specific antibody against A. paragallinarum serotype C in chickens and 37.5% of them were protected post-challenge with A. paragallinarum serotype C strain 668. The results indicated a prospect of applications of the epitope in the development of peptide vaccines against poultry infectious coryza.

关 键 词：副鸡禽杆菌 表位 噬菌体展示 

分 类 号：S852.612[农业科学—基础兽医学]