Yeast surface display system of infectious hematopoietic necrosis virus glycoprotein

Glycoprotein is the most important surface antigen of infectious haematopoietic necrosis virus (IHNV), and it has been served as the major gene for virus detection and vaccine development. The specific PCR primers for IHNV glycoprotein gene replication were designed based on the plasmid containing IHNV-Sn1203 glycoprotein gene. The PCR product was ligated to yeast surface display vector pYD1 and the recombinant plasmid pYD1-G was transformed into EBY100 cell. The transformed EBY100/pYD1-G cell was induced by galactose and the protein expression was detected by cell immunofluorescence, Western Blotting and flow cytometry. The cell immunofluorescence result showed that the transformed EBY100 cell presented a red fluorescence, and Western blotting analysis obtained a specific glycoprotein band. Both of these two results indicated that the glycoprotein was successfully displayed on the surface of yeast cell. The flow cytometry analysis showed that the glycoprotein expression was increased with the galactose induced time, and the maximum expression was obtained at 48 h. The successful display of glycoprotein on the yeast surface laid a foundation for live oral vaccine development.