• Journal of fish pathology
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• v.24 no.2
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• pp.75-84
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• 2011

The amino acid sequence of glycoprotein of Korean VHSV isolate (KR'01-1) was analyzed using the DNAStar Protean system. Based on the flexibility, hydrophilicity, antigenic index and surface probability, three regions (Gp1, Gp2 and Gp3) were selected as potential antigenic determinants. Three oligopeptides containing the amino acid sequences of the three regions were synthesized and polyclonal antibodies were raised against them. The activities of the antibodies were analyzed by Western blotting and virus neutralization test. The results showed that antibodies raised against oligopeptides Gp1 and Gp2 neutralized the infectivity of VHSV, suggesting that they can be possible candidates for subunit vaccines against VHS diseases in olive flounder.

• Proceedings of the Korean Society of Fisheries Technology Conference
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• 2000.05a
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• pp.446-446
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• 2000

Specific polyclonal and/or monoclonal antibodies (MAbs) to immunoglobulins (Igs) and their subunits have proved to be valuable tools in immunological research and in immunological assays. In this study, we developed and characterized MAbs against flounder serum Igs. To obtain the pure flounder serum Igs, mouse IgG (mIgG) was immunized to flounder. Flounder Igs were purified by using mIgG-agarose affinity column chromatography. The structure of purified flounder Ig was observed, on denatured SDS-PAGE, to be composed of two heavy chains (77 and 72 kd) and two light chains (28 and 26 kd). MAbs were produced by fusion of myeloma cells (SP2/0) with Balb/c mouse spleen cells previously primed with the flounder Igs. Finally, three hybridoma clones, FIM 511, FIM 519 and FIM 562 were established to recognize both 2 heavy chains, 26 kd of light chain and 28 kd of light chain, respectively. On the other hand, the flounder immune sera collected on the weekly basis were tested on ELISA and immunoblot analysis whether boosting effect is present in flounder humoral immune system. As a result, the secondary immune response in flounder was ascertained on ELISA, but not on immunoblot analysis. Further, we observed an alteration of serum protein levels following immunization. Our MAbs and basic information on flounder humoral immune system obtained in this study will be helpful to control and monitor the efficiency of fish vaccines and therapeutic process of flounder diseases.

• Journal of Fisheries and Marine Sciences Education
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• v.26 no.6
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• pp.1366-1372
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• 2014

This study was conducted to select the media for the formalin killed vaccine (FKC) production of Vibrio harveyi and its application for olive flounder, Paralichthys olivaceus. For this, we have investigated the immune effects of Vibrio harveyi FKC vaccines grown in 3 different media i.e. Tryptic Soy Broth (TSB), TSB containing 2-2'-dipyridyl (TSB-D), Brain Heart Infusion Broth (BHIB) on the production of agglutinating antibody and protection against Vibrio harveyi in olive flounder. Additionally, a dual vaccine was prepared by combining Streptococcus parauberis vaccine to V. harveyi vaccine and its efficacy was also analyzed with the determination of optimal administration dosage. Consequently, olive flounder immunized with FKC grown in TSB-D showed the same protection with the vaccine grown in BHIB and the optimal dose of the vaccine was 10mg/kg of body weight. Indeed the dual vaccine showed higher agglutination titer and protection than control fish. The optimal dose for dual vaccine was 10mg/kg body weight for each of two vaccines.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.18 no.5
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• pp.464-470
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• 1985

Some eels Anguilla japonica, ranging from 16 to 23 g in their weight(average: 20 g), were sampled at the private eel farming company equipped with water recycling system, located at Kimhae city, Kyungnam Province, Korea. Three kinds of vaccine were prepared with Vibrio anguillarum (EPM-8406) isolated at National Fisheries University in Korea for the immune response experiment against eels; those vaccines were made by inactivating the strain with $0.3\%$ formalin for 24 hrs at $25^{\circ}C$, heating for 3 mins or for 15 mins at $121^{\circ}C$, respectively. The various optimal vaccination conditions for the control of vibriosis in the fish were investigated based on the cultivation temperature, vaccination concentration and booster effect. The maximum titer rapidly increased with higher temperature up to $23^{\circ}C$, but there were little differences between $23^{\circ}C\;and\;28^{\circ}C$. The formalin-killed vaccine showed good efficacy at the injection concentration of above $10^8$ cells per fish and little effect at the below $10^7$ cells. The booster effect on the vaccination showed good efficacy above twice-injections with little difference between the numbers of injection.

• Journal of Life Science
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• v.30 no.4
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• pp.402-409
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• 2020

Nervous necrosis virus (NNV) contains a bi-segmented viral genome, RNA1 (3.4 kb, RdRp), and RNA2 (1.4 kb, capsid protein) in a small particle (25 nm). Despite its extremely compact size, NNV has caused serious damage by infecting approximately 120 fish species worldwide since it was first reported in the late 1980s. In order to minimize the damage caused by NNV infection and develop effective vaccines, it is necessary to understand the intra cellular signaling system according to NNV infection. NNV infection induces cell cycle arrest at the G1 phase via the p53-dependent pathway to use the cellular system for its replication. Otherwise, host cells recognize NNV infection through the RIG-1-like receptor (RLR) signaling pathway to control the virus and infected cells, and then ISGs required for antiviral action are activated via the IFN signaling pathway. Moreover, apoptosis of infected cells is triggered by the unfolded protein response (UPR) through ER stress and mitochondria-mediated cell death. Cell signaling studies on the NNV infection mechanisms are still at an early stage and many pathways have yet to be identified. Understanding the various disease-specific cellular signaling systems associated with NNV infection is essential for rapid and accurate diagnosis and vaccine development.