Capacity of the structural protein VP1 of foot-and-mouth disease virus potentially accommodating insertion of different foreign tags

Abstract:

[Objective] To identify the capacity that the structural protein VP1 of foot-and-mouth disease virus potentially accommodate insertion of different foreign tags, we constructed recombinant FMDVs containing foreign tags using FMDV reverse genetics system. [Methods] Using overlap extension PCR method, we introduced V5, TC12, KT3, 3×FLAG tag genes into G-H loop of VP1 capsid protein of FMDV. Linearized recombinant plasmids were transfected into BSR/T7 cells expressing T7 RNA polymerase to rescue the recombinant viruses. The recombinant viruses were analyzed by RT-PCR, indirect immunofluorescence, plaque phenotype and one-step growth curves. [Results] We successfully rescued the recombinant FMDVs expressing V5 and KT3 tag but could not rescue the recombinant FMDV containing TC12 and 3×FLAG tags. The introduction of V5 and KT3 tags both affected the replication capacity of FMDV. [Conclusion] Our results will lay foundations to study marker vaccine and FMDV vector in future.

Key words:foot-and-mouth disease virus;structural proteins;foreign tags