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草鱼呼肠孤病毒(GCRV)是引起我国大面积草鱼幼鱼出血病暴发的主要病原,其外衣壳蛋白VP5和VP7在病毒入侵宿主细胞过程中起着至关重要的作用。研究以原核表达的VP7、全长VP5、VP5的N端片段及C端片段为靶蛋白,利用已构建的噬菌体展示单链抗体文库进行淘选。经过3轮淘选后,共获得7个针对VP7、VP5、VP5N和VP5C的单链抗体。经过验证,识别原核表达的VP7的两个单链抗体能够成功识别天然GCRV病毒。此结果对于进一步研究GCRV与宿主细胞的相互作用机理奠定了基础。
Grass carp reovirus (GCRV) is the major causative agent of outbreaks of large-scale grass carp juveniles in China. Its capsid proteins VP5 and VP7 play crucial roles in virus invasion of host cells. The prokaryotic expression of VP7, full-length VP5, VP5 N-terminal fragment and C-terminal fragment as the target protein, the use of constructed phage displayed single chain antibody library panning. After 3 rounds of panning, a total of seven single chain antibodies against VP7, VP5, VP5N and VP5C were obtained. After validation, two single chain antibodies that recognize prokaryotic expressed VP7 are able to successfully recognize the native GCRV virus. This result laid the foundation for further study of the interaction mechanism between GCRV and host cells.