Aim: To construct a recombinant strain BL21(DE3)(pET-28a-OmpS(2);), and obtain a genetic engineering vaccine to provide protective immunity against diseases caused by Edwardsiella tarda.
Methods: According to the GenBank sequences (GenBank Accession No. AY078509), one pair of primers was designed and the outer membrance protein gene (OmpS(2);) of Edwardsiella tarda HB01 was amplified by PCR. The OmpS(2); gene was cloned in pET-28a vector and transformed into Escherichia coli BL21(DE3). The OmpS(2); protein was highly expressed when the recombinant strain BL21 (DE3) (pET-28a-OmpS(2);) was induced by IPTG. The expressed protein was 47 kD as estimated by 150 g/L SDS-polyacrylamide gel electrophoresis (SDS-PAGE).
Results: The immunogenicity of the expressed OmpS(2); protein was confirmed by Western blotting. Mice and Paralichthys olivaceus were immunized with the genetic engineering vaccines of Edwardsiella tarda and Aeromonas hydrophila, showing promise that all these vaccines have a high protective ability. And the protective ability to Edwardsiella tarda and Aeromonas hydrophila in Paralichthys olivaceus respectively reached 70% and 80%.
Conclusion: The recombinant strain BL21 (DE3)(pET-28a-OmpS(2);) could be candidate of genetic engineering vaccine to provide protective immunity against diseases caused by Edwardsiella tarda.
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