Tandem and Prokaryotic Expression of VP1 Gene of Foot-and-Mouth Disease Virus Type C

Objective To construct a prokaryotic expression system for the VP1 gene of foot-and-mouth disease virus(FMDV) type C and study the immunoreactivity of expressed product.Methods Synthesize three epitopes of VP1 gene of C-S8C1 strain of FMDV and clone into pMD18-T vector.Digest the recombinant plasmid with restriction endonuclease and link the digested fragments to construct a double copy gene fragment VP1C.A recombinant plasmid pETCKS-VP1C was constructed by inserting VP1C gene fragment into prokaryotic expression vector pET-CKS and transformed to E.coli BL21 for expression.Identify the expressed product by SDS-PAGE and analyze its immunoreactivity by Western blot.Purify the expressed protein,in a form of inclusion body,by washing.Results VP1C gene was expressed in E.coli.The expressed product was recognized by the standard antiserum against FMDV type C and contained 45% of total somatic protein.The purity of recombinant VP1C protein after purification by the washing of inclusion body reached 84.1%.Conclusion A prokaryotic expression system for the VP1 gene of FMDV type C was successfully constructed.The expressed VP1C showed good immunoreactivity and might be used as a candidate gene for detecting FDMV type C.