传染性支气管炎病毒抗原模拟表位在大肠杆菌鞭毛上的展示

The Expression of Infectious Bronchitis Virus Mimotopes on E.coli Flagellin

目的:进一步验证通过噬菌体肽库筛选得到的2个传染性支气管炎病毒(IBV)抗原模拟表位在脱离噬菌体后仍具有与IBV抗体结合的生物学活性。方法:应用基因工程技术合成2个IBV抗原模拟表位(KSPKHSSSALHF和SⅡQMNLHR PTS)的编码碱基序列,将其分别插入质粒pFliTrx,构建重组展示载体p1和p2,并分别转化大肠杆菌GI826,形成展示IBV模拟抗原表位肽的重组菌F1和F2,进行体外抗原抗体反应。结果:体外抗原抗体反应试验表明,重组菌F1与F2可以与IBV阳性鸡血清特异性结合。结论:提示得到的2个抗原模拟表位在脱离噬菌体后,仍具有与IBV抗体结合的生物学活性,在研制IBV新型疫苗和诊断试剂方面具有潜在的应用价值。

Objective： To attest if the two infectious bronchitis virus（IBV） mimotopes which were obtained from phage display peptide library still have specific affinity to corresponding IBV antibodies when they were departed from phages. Methods： The DNA fragments that coded the peptide sequences KSPKHSSSALHF and SIIQMNLHRPTS were cloned into the expression vector pFliTrx by means of genetic engineering technique, respectively. The recombinant plasmids pl and p2 were constructed and transformed into an expression host E.coli GI826 respectively. And two recombinant E.coli F1 and F2 which displaying the IBV mimotopes on the bacterial flagellum were constructed successfully. Results： The recom- binant bacteria showed actual positive reactivity with IBV antiserum by antigen-antibody recognition in vitro. Conclusion： The results indicate that two IBV mimotopes still have specific affinity to corresponding IBV antibodies when they were departed from phages. It may provide a foundation for developing a new type vaccine and diagnostic reagents for IBV.