| Edwardsiella tarda has become an important fish pathogenicity bacterium, which has seriously harmed to aquacuture around the world. Based on the reverse vaccinology, genome sequence of E. tarda EIB202 was analyzed by a variety of bioinformatic methods and 275 candidate antigens were selected in this work. Among them,193 antigens were successfully cloned and 85 recombinant proteins were highly expressed in Escherichia coli and successfully purified.16 antigens led to a high relative percent survival (RPS> 50%) when challenged with E. tarda EIB202.Meanwhile, combined vaccination was performed with GAPDH and FlgD which possessed high relative percent survivals and were different in antigen-presenting. Immune protections were carried out on experimental animal zebrafish and clinical animal turbot. Mechanism of the immune response was also studied with RT-qPCR. And the results showed that the humoral immunity and cellular immunity were obviously induced in the vaccinated turbot with the mixture of rGAPDH and rFlgD. Thus it provided a good idea for multiple antigens combined vaccination and will be helpful for the vaccine design of aquaculture in the future. |
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