Rescue and identification foot-and-mouth disease virus Asia1/JS/China/2005 strain with Arg-Gly-Asp RGD receptor recognition site

Abstract:

Abstract: [Objective] To construct an infectious full-length cDNA clone of Asia1/JS/China/2005 strain with Arg-Gly-Asp (RGD) receptor recognition site. [Methods] We constructed foot-and-mouth disease virus type Asia1 full-length cDNA clone pFMDV-RGD by using site-directed mutagenesis. The plasmid pFMDV-RGD contained the desired mutation. The plasmids pFMDV-RGD were linearized with NotI enzyme. Linearized plasmid and pcDNAT7P plasmids expressing T7 RNA polymerase cotransfected into BHK-21 cells to rescue FMDV-RGD. [Results] We constructed FMDV Asia1/JS/China/2005 strain full-length cDNA clone with Arg-Gly-Asp receptor recognition site by sequence. We obtained rescued virus by plasmid cotransfection. The results of sequencing, indirect immunofluorescence, electron microscope and sulk mice pathogenicity analysis showed foot-and-mouth disease virus containing Arg-Gly-Asp receptor recognition site was successfully rescued. [Conclusion] The results lay a foundation for further study of biology characteristic diversity of rescued virus with Arg-Gly-Asp and Arg-Asp-Asp (RDD) receptor recognition site.