Generation of a temperature-sensitive Edwardsiella tarda mutant and its potential as a prophylactic vaccine in olive flounder (Paralichthys olivaceus)

Journal of Applied Microbiology
S H ChoiKi Hong Kim

Abstract

The aim of this study was to generate temperature-sensitive Edwardsiella tarda mutant and to evaluate potential of the mutant as a vaccine in olive flounder (Paralichthys olivaceus). A temperature-sensitive E. tarda mutant was generated by replacement of alr gene promoter with cI857-λP(R) promoter system plus another CI857 expression cassette that was driven by a constitutive promoter of E. tarda (EtPR C28-1). Growth of the mutant strain was not different to that of wild-type E. tarda under conditions of culture at 39°C. However, growth of the mutant strain was retarded by culturing at 25 or 20°C without d-alanine. To further inhibit leakage of λP(R) promoter, the mutant strain was transformed with a vector harbouring an EtPR C28-1-driven cI857 cassette (pEtPR-cI857), which resulted in more limited growth compared to the mutant without the plasmids. The level of alr gene transcription in the mutant E. tarda and the mutant harbouring pEtPR-cI857 was well coincided with the result of bacterial growth. Olive flounder fingerlings immunized with the E. tarda mutant showed decreased mortality, and a boost immunization induced 100% protection against E. tarda infection. The protection rate of fish was proportional to the serum aggluti...Continue Reading

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Citations

Nov 12, 2015·Frontiers in Immunology·Hetron Mweemba Munang'anduØystein Evensen

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