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KMID : 0545120050150040873
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Journal of Microbiology and Biotechnology
2005 Volume.15 No. 4 p.873 ~ p.879
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Development of DNA Vaccine Against Red Sea Bream Iridovirus (RSIV)
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Park SJ
Seo HJ/Son JH/Kim HJ/Kim YI/Kim KH/Nam YK/Kim SK
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Abstract
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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 105.5 TCID50/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 RSIV 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.
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