Journal of Microbiology and Biotechnology

  • 제15권4호
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  • Pages.873-879
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  • 2005
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  • 1017-7825(pISSN)
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  • 1738-8872(eISSN)
한국미생물·생명공학회 (The Korean Society for Microbiology and Biotechnology)
권호 표지

Development of DNA Vaccine Against Red Sea Bream Iridovirus (RSIV)

  • PARK SO-JIN (Department of Biotechnology and Bioengineering, Pukyong National University) ;
  • SEO HYO-JIN (Interdisciplinary Program of Marine Biotechnology, Pukyong National University) ;
  • SON JEONG HWA (Department of Biotechnology and Bioengineering, Pukyong National University) ;
  • KIM HYOUNG-JUN (Department of Aquatic Life Medicine, Pukyong National University) ;
  • KIM YUN-IM (Department of Biotechnology and Bioengineering, Pukyong National University) ;
  • KIM KI-HONG (Department of Aquatic Life Medicine, Pukyong National University) ;
  • NAM YOON-KWON (Department of Aquaculture, Pukyong National University) ;
  • KIM SUNG-KOO (Interdisciplinary Program of Marine Biotechnology, Pukyong National University)
  • 발행 : 2005.08.01
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초록

Red sea bream iridovirus (RSIV) obtained from infected rock bream was propagated by Bluegill fry-2 (BF-2) cell culture. The virus titer was determined as $10^{5.5}\;TCID_{50}/ml$ on confluent BF-2 cell monolayers. The integrin binding site of ORF 055L of infectious spleen and kidney necrosis virus (ISKNV) was selected for the construction of a primer to obtain the RSIV ORF 055L gene. The genes were amplified using RSIV gene lyzate by PCR. The homologies of the ORF 055L sequence of RSIV with ISKNV and rock bream iridovirus (RBIV) were approximately $96\%$ and $100\%$, respectively. DNA vaccine was constructed by cloning the ORF 055L of RSN into pcDNA 3.1 (+), containing a cytomegalovirus (CMV) promoter. For antibody production, pcDNA-055 DNA vaccine was injected to BALB/c mice. The production of antibodies against pcDNA-055 DNA vaccine was confirmed by the Western blot analysis. The antibodies produced by the pcDNA-055 DNA vaccine showed efficacy to neutralize the RSIV in the neutralization test in BF-2 cell culture.

키워드

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