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. 2022 Jul 29;13:932722. doi: 10.3389/fimmu.2022.932722

Figure 2.

Figure 2

Schematic diagram of the yeast vaccine and experimental strategy. (A) pYD1-ORF25 recombinant plasmids displayed on the yeast cell surface through attachment of Aga2 to Aga1 via disulfide bonds. (B) Positive clones of pYD1-ORF25 identification by agarose gel electrophoresis. (C) Immunofluorescence assay of EBY100/pYD1-ORF25 fusion protein Expression. Scale bars, 20 μm. (D) Strategy to vaccinate gibel carp. Briefly, vaccine and bo-vaccine fish were orally administered with CyHV-2 yeast vaccine at 1, 2, and 3 days (vaccine) and 1, 2, 3, 18, 19, and 20 days (booster vaccine), respectively. Take samples at the 10, 17, 27, and 34 days to test the safety of the yeast vaccine (cumulative survival after vaccination and morphological changes of gut), body fluid biochemistry indices, and immune-related gene expression. Challenge with CyHV-2 at the 41 days to test intestinal pathological changes, viral load, and relative percent survival.