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USING OF METHOD ELECTROPORATION FOR TRANSFER OF DNA VNN (VIRAL NERVOUS NECROSIS) VACCINES IN GROUPER FISH

Wiwien Mukti A.1), Sri Murtini2), Surachmi2), Alimuddin3)
 
1Brackishwater Aquaculture Development Center, Situbondo, East Jawa. Indonesia.
3Faculty of Veterinary Science, IPB, Bogor, Indonesia
2Department of Aquaculture, Faculty of Fisheries and Marine Science,
Bogor Agricultural University (IPB), Bogor 16680, Indonesia

Viral nervous necrosis (VNN) was a disease that often attacks the grouper. One way of preventing infectious diseases with DNA VNN vaccination. In this study, the method used to transfer DNA VNN vaccine was electroporation using a series of electric shocks to open the pores of fish sperm cell membranes, and causing the DNA vaccine can enter the cell. The purpose of this study was to examine the vaccination potential of DNA VNN in larvae through electroporation method of sperm and do confirmation test ability of DNA VNN vaccines with different promoter on grouper. In These research used machines BioRad electroporation system, at a voltage of 50 volts, time of shock 30 ms, the number of shock 5, shock interval of 0.1 μs, and the size of cuvette 0.4 cm electroporation of sperm using DNA vaccines VNN was containing different promoter construction, namely  pJfKer-PCV, and pmBA-PCV. Parameters measured were mRNA of VNN coat protein in larvae the age of one day (D1), three day (D3) and seven day (D7) as a regulator of the immune system on the viability of grouper. The results showed that the DNA vaccine VNN delivery on sperm effective using electroporation methods, indicated by have been detected of mRNA gene expression VNN at larvae age one day (D1). The product of DNA PCR was shown at size 195 bp. Based on the results of this study concluded that effective electroporation method and can be used in DNA vaccine delivery VNN through sperm on grouper.

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