Development of DNA vaccines for fish

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We have constructed fish DNA vaccines using G-protein genes of hirame rhabdovirus (HIRRV), viral haemorrhagic septicemia (VHS) virus, major capsid protein gene and attachment protein gene of red seabream iridovirus, three different major antigenic protein genes of Pasteurdla piscidda, and Antigen85 genes of Mycobacterium sp. The expression of these genes was controlled under a virus promoter for expression in fish cells. We also constructed a DNA adjuvant using Japanese flounder interleukin-1 gene.The DNA vaccine which encoded the G-protein gene of KERRY had high protective activity against HERRV infection when this DNA vaccine was injected over 1|jg to 1 to 2 g Japanese flounder. The DNA vaccines which encoded the major casid protein gene or attachment protein gene also have protective activity against red seabream iridovirus. However, in case of iridovirus DNA vaccine, the protective activities were not so high and the RPS values were about 50 %.We have cloned and characterized some Japanese flounder gene promoters for the construction of an all fish DNA vaccine vector. We cloned the Japanese flounder complement component C3, keratin, β-actin, gelatinase and TNF-ct gene promoters on up stream of the green fluorescent (GFP) gene. We constructed transgenic zebrafish for characterization of the promoter activities. The promoter activity analysis suggest that the keratin gene promoter was most suitable for DNA vaccine because its promoter activity was detected through out the body of zebrafish.We developed techniques for quantification of rnRNA from a number of defense and immune-related gene of Japanese flounder.