利用荧光标记的T7噬菌体研究配体/受体的相互作用

将鸡传染性法氏囊病病毒(IBDV)衣壳蛋白VP2展示到T7噬菌体表面,以FITC标记纯化的重组噬菌体,通过荧光显微镜观察与流式细胞仪检测,研究标记噬菌体与病毒受体细胞--法氏囊B细胞的相互作用.结果展示有IBDV VP2蛋白的噬菌体经FITC标记后仍然具有与受体细胞结合的特性,荧光显微镜下可见绿色荧光,流式数据显示其平均荧光强度明显高于阴性对照,且IBDV疫苗株TAD可明显阻断其结合.由此得出结论,FITC标记与噬菌体展示技术相结合,可进行配体/受体间相互作用的研究.

Use of Fluoresceinated Recombinant Phage to Study the Receptor/Ligand Interaction

VP2 protein of infectious bursal disease virus(IBDV) was displayed on T7 phage surface, and this recombinant phage was then purified and labeled with FITC. The interaction between fluorescigenic phage and IBDV host cells was detected by fluorescent microscope and flow cytometer(FCM). Data shows that after displayed on labeled T7 phage suface, VP2 protein still remains its binding activities with IBDV host cells, and this interaction can be blocked by IBDV vaccine strain TAD in a does dependent manner, while no interaction was observed in negative control. The described method should find widespread application in the rapid in vivo research of interactions between ligands and receptors.