ESTABLISHMENT OF YEAST EXPRESSION SYSTEM FOR SPRING VIRERNIA OF CARP VIRUS GLYCOPROTEIN

In this study, open reading frame (ORF) of glycoprotein (1530 bp) was amplified by using RNA extracted from Spring Virernia of Carp Virus (SVCV). The SVCV G ORF was cloned into pYD1 vector to construct a recombi-nant plasmid pYD1-G and then transformed into competent yeast cells EBY100, and positive colonies were screened by colony PCR. The expression of G gene was induced by 2% glucose and detected by cell immunofluorescence and flow cytometry. The immunofluorescence staining observed increased EBY100-pYD1-G signal of induced yeast cells with increased induction time. Flow cytometry analysis observed significantly increased fluorescence intensities in prolonged induced EBY100-pYD1-G cells (P<0.05). These results indicated the SVCV G protein has been successfully expressed and localized on the surface of yeast cell. This study laid a foundation for the novel oral vaccine development against SVCV infection in carps in future.