【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。更多还原

【Abstract】 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 McAb,which could be competitively inhibited by A.paragallinarum.One of the peptide sequences,YSPHQWWLSGAV,which was itself expressed with the flagella on the Escherichia coli GI826 cell surface was selected and inserted into the multiple cloning sites of pFliTrx vector.Intramuscular injection of the epitope-containing 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.更多还原