Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Evaluation of Optimal Culture Conditions for Recombinant Ghost Bacteria Vaccine Production with the Antigen of Streptococcus iniae GAPDH

  • Ra, Chae-Hun (Departments of Biotechnology, Pukyong National University) ;
  • Kim, Yeong-Jin (Departments of Biotechnology, Pukyong National University) ;
  • Park, So-Jin (Natural Products Research Institute, Binex Co. Ltd) ;
  • Jeong, Chang-Wha (Natural Products Research Institute, Binex Co. Ltd) ;
  • Nam, Yoon-Kwon (Departments of Aquaculture, Pukyong National University) ;
  • Kim, Ki-Hong (Departments of Aquatic Life Medicine, Pukyong National University) ;
  • Kim, Sung-Koo (Departments of Biotechnology, Pukyong National University)
  • Published : 2009.09.30
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Abstract

For the production of ghost bacteria vaccine to prevent the streptococcal disease in aquaculture fish species, a double cassettes vector was constructed and cloned in Escherichia coli DH5${\alpha}$. Ghost bacteria vaccine production from Escherichia coli DH5${\alpha}$/pHCE-InaN-GAPDH-Ghost 37 SDM (SIG) was maximized at a glucose concentration of 1 g/l, agitation of 300 rpm, and aeration of 1 vvm. The maximal efficiency of ghost bacteria formation was obtained at the mid-exponential phase ($OD_{600}=2.0$) with the concentration of 0.77 g/l for SIG. The molecular mass of GAPDH was detected at 67 kDa with the insoluble fraction, by SDS-PAGE and Western blot. The protective efficacy of ghost bacteria vaccine was evaluated by challenge test using olive flounder. The cumulative mortalities of the positive control, formalin-killed cell (FKC) vaccine, and SIG vaccine immunized groups were 91%, 74%, and 57%, respectively. These results suggest that SIG vaccine showed efficacy as a vaccine and had a higher potential to induce protective antibodies than did FKC vaccine.

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References

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