• Journal of fish pathology
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• v.22 no.3
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• pp.353-366
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• 2009

We constructed a rock bream Oplegnathus fasciatus leukocyte cDNA library and a total of 795 expressed sequence tag (EST) clones were generated. Gene annotation procedures and homology searches of the sequenced ESTs were locally done by BLASTX for amino acid similarity comparisons. Of the 795 EST clones, 491 different ESTs showed significant homology to previously described genes while 304 ESTs were unidentified, hypothetical, or unnamed proteins. Encoding 121 different sequences were identified as putative bio-defense genes or genes associated with immune response.

• Journal of fish pathology
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• v.21 no.2
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• pp.129-137
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• 2008

We constructed a black rockfish (Sebastes schlegeli) leukocyte cDNA library and a total of 386 expressed sequence tag (EST) clones were generated. Gene annotation procedures and homology searches of the sequenced ESTs were locally done by BLASTX for amino acid similarity comparisons. Of the 386 EST clones, 199 different ESTs showed significant homology to previously described genes while 97 ESTs were unidentified, hypothetical, or unnamed proteins. Encoding 38 different sequences were identified as putative bio-defense genes or genes associated with immune response.

• Journal of fish pathology
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• v.33 no.2
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• pp.103-109
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• 2020

In order to use nervous necrosis virus (NNV) virus-like particles (VLPs) as a delivery tool for heterologous antigens or plasmids, we attempted to produce red-spotted grouper nervous necrosis virus (RGNNV) VLPs displaying a partial region of viral hemorrhagic septicemia virus (VHSV) glycoprotein at the surface and VLPs that are harboring DNA vaccine plasmids within the VLP. A peptide encoding 105 amino acids of VHSV glycoprotein was genetically inserted in the loop region of NNV capsid gene, and VLPs expressing the partial part of VHSV glycoprotein were successfully produced. However, in the transmission electron microscope analysis, the shape and size of the partial VHSV glycoprotein-expressing NNV VLPs were irregular and variable, respectively, indicating that the normal assembly of capsid proteins was inhibited by the relatively long foreign peptide (105 aa) on the loop region. To encapsulate by simultaneous transformation with both NNV capsid gene expressing plasmids and DNA vaccine plasmids (having an eGFP expressing cassette under the CMV promoter), NNV VLPs containing plasmids were produced. The encapsulation of plasmids in the NNV VLPs was demonstrated by PCR and cells exposed to the VLPs encapsulating DNA vaccine plasmids showed fluorescence. These results suggest that the encapsulation of plasmids in NNV VLPs can be done with a simple one-step process, excluding the process of disassembly-reassembly of VLPs, and NNV VLPs can be used as a delivery tool for DNA vaccine vectors.

• Journal of Microbiology and Biotechnology
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• v.32 no.1
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• pp.64-71
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• 2022

The discarding of wastes into the environment is a significant problem for many communities. Still, food waste can be used for lactic acid bacteria (LAB) growth. Here, we evaluated three growth media equivalent to de Mann Rogosa Sharpe (MRS), using apple bagasse, yeast waste, fish flour, forage oats, and cheese whey. Cell-free supernatants of eight LAB strains were tested for antimicrobial activity against nine indicator microorganisms. The supernatants were also evaluated for protein content, reducing sugars, pH, and lactic acid concentration. Cell-free supernatants from fish flour broth (FFB) LAB growth were the most effective. The strain Leuconostoc mesenteroides PIM5 presented the best activity in all media. L. mesenteroides CAL14 completely inhibited L. monocytogenes and strongly inhibited Bacillus cereus (91.1%). The strain L. mesenteroides PIM5 consumed more proteins (77.42%) and reducing sugars (56.08%) in FFB than in MRS broth (51.78% and 30.58%, respectively). Culture media formulated with agroindustrial wastes positively improved the antimicrobial activity of selected LAB, probably due to the production of antimicrobial peptides or bacteriocins.

• Journal of fish pathology
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• v.35 no.1
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• pp.121-128
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• 2022

Recent studies revealed that histone proteins are involved in innate immune responses during pathogen invasion as well as DNA packing. This study characterized the histone genes (H2A.V) of sevenband groupers and analyzed gene expression in NNV-infected sevenband groupers. The open reading frame (ORF) of H2A.V is 387 bp which encoded 128 amino acid residues. The deduced amino acid sequence of H2A.V harbor a highly conserved domain for H2A/H2B/H3 and H2A_C binding domain. Quantitative real-time PCR analysis showed that H2A.V had a high gene expression level in the brain and blood after being NNV-infected. An increase in extracellular histone protein in the blood has been identified as a biomarker for vascular function in humans. More research is required to understand histone's immune response at the protein level or in aquatic animals.

• Journal of Life Science
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• v.26 no.12
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• pp.1360-1366
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• 2016

Heat shock proteins (HSPs) are induced in response to various physiological or environmental stressors. However, the transcriptional activation of HSPs is regulated by a family of heat shock factors (HSFs). Fish models provide an ideal system for examining the biochemical and molecular mechanisms of adaptation to various temperatures and water environments. In this study, we examined the pattern differentials of heat shock factor 1 (HSF1) and expression of heat shock protein 70 (HSP70) in response to thermal stress in goldfish and mouse hepatocyte cultures by immune-blot analysis. Goldfish HSF1 (gfHSF1) changed from a monomer to a trimer at $33^{\circ}C$ and showed slightly at $37^{\circ}C$, whereas mouse HSF1 (mHSF1) did so at $42^{\circ}C$. This experiment showed similar results to a previous study, indicating that gfHSF1 and mHSF1 play different temperature in the stress response. We also examined the activation conditions of the purified recombinant proteins in human HSF1 (hmHSF1) and gfHSF1 using CD spectroscopy and immune-blot analysis. The purified recombinant HSF1s were treated from $25^{\circ}C$ to $42^{\circ}C$. Structural changes were observed in hmHSF1 and gfHSF1 according to the heat-treatment conditions. These results revealed that both mammal HSF1 (human and mouse HSF1) and fish HSF1 exhibited temperature-dependent changes; however, their optimal activation temperatures differed.

• Journal of Life Science
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• v.25 no.2
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• pp.214-222
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• 2015

Aeromonas hydrophila is the most common water fish pathogen and cause diseases such as hemorrhagic septicemia, dropsy, ulceration and asymptomatic septicemia. A. hydrophila secretes many extracellular products (ECPs) which contribute to effective infection, wide distribution and great adaptability to environmental changes. Crude ECPs of A. hydrophila CK257, a strain used in this study, exhibits a toxic activity to the animals including mouse, rabbit and fish. Toxic symptoms were indicated by tissue damage and skin injuries in animal. When ECPs were subcutaneously injected to animals, skin damages were observed, appearing like necrosis. Preliminary research demonstrated that the active factors are protein component. The crude ECPs were collected after ammonium sulfate precipitation of cell-free culture supernatant. ECPs were fractionated with the use gel filtration chromatography. Five ECP fractions were obtained, of which one fraction was found to be toxic to goldfish. MALDI-TOF analyses provided two interesting proteases called M35 and M28. Both M35 and M28 are known as metalloprotease. Accordingly, proteins in an active fraction exhibited caseinolytic activity. These proteins were difference of caseinolytic activity under different metallic ions. Also active fraction has elastolytic activity. These results suggested that peptidase M28 and M35 may be a candidate factor for tissue necrosis activity about infection with A. hydrophila.

• Korean Journal of Ichthyology
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• v.18 no.2
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• pp.65-77
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• 2006

Gene transcripts potentially responsive to the heat stress were surveyed by cDNA microarray analysis in mud loach (Misgurnus mizolepis). Transcriptional profiles of hepatic tissue in the fish exposed to either $23^{\circ}C$ or $32^{\circ}C$ for 4 weeks were compared each other by 3 replicated hybridization assays using 1,124 unigene clones selected from mud loach liver expressed sequence tags (ESTs). A total of 93 clones showed the substantially increased mRNA levels (>2-fold) in $32^{\circ}C$-exposed group when compared in $23^{\circ}C$control group. It includes various enzymes and proteins involved in energy pathway, protease/protein metabolisms, immune/antioxidant functions, cytoskeleton/cell structure, transport and/or signal transduction. Maximum level of increase was up to 15-fold relative to $23^{\circ}C$ treatment. Heat exposure also resulted in the significant decrease (less than 50% relative to $23^{\circ}C$-exposed fish) of the transcriptional activities in 85 genes. Besides the above categories, yolk protein (vitellogenin) and ribosomal proteins were notably down regulated in the fish exposed to heat stress. A number of novel gene transcripts were also detected in both up-regulated and down-regulated groups.

• Journal of the Korean Society of Food Science and Nutrition
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• v.20 no.5
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• pp.455-460
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• 1991

In order to identify hard distinct 12 fish species(shiba shrimp Metapenaeus joyneri, fleshy shrimp Penaeus orientalis, ridgetail prawn Palaemon carinicauda, yellow croaker Pseudosciaena manchurica, croaker Niber albiflora, Colichthyes fragilis, brown sole Limanda herzensteini, frog flounder Pleuronichthys cornutrs, Areliscus rhomaleus, stone flounder Kareius bicoloratus, harvest fish Pampus argenteus, flag fish Goniistius zonatus) by seeing with naked eye in Kunsan coastal area, sarcoplasmic protein in the supernatant was used for isoelectric focusing. For getting supernatant, fish muscle tissue was blended with two times deionized water and centrifuged (at $4^{\circ}C$, 12,000rpm for 15min). Isoelectric focusing of sarcoploasmic protein carried out on a LKB Multiphor II using polyacrylamide gel plate (2mm thickness, pH $3.5~10^{\circ}C$, pH 5~8 gradient, at $10^{\circ}C$ for 1.5, 3 hours). In case of uncertain protein pattern, pH gradient was modified to narrow pH gradiet, and excuted 2-D electrophoresis using conventional polyacrylamide gel electrophoresis. Most of fishes except yellow croaker and Collichthyes fragilis were distingushed by isoelectric focusing. The protein maps of 2-D electrophoresis for analyzing two protein bands at aimilar positions(pH 5, 6) between the two fish species showed the diffeences of the estimated molecular weights, 11,700(pH5.0) and 87,000(pH6.0)

• Korean Journal of Environmental Biology
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• v.33 no.1
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• pp.7-18
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• 2015

In this study, we studied the effect of dissolved cement powder on Carassius auratus by analysis of the morphophysiological changes. The gill exposed to dissolved cement powder showed the thickened primary lamellae and the activity of chloride cells and mucous cells was also significantly increased and the proliferation, separation and clubbing of gill filament was observed in the secondary lamellae. In the kidney tissue, the space in Bowman's capsule was widen and the arrangement of dermis was irregular due to the thinned epidermis in the integument tissue. The activities of antioxidant enzymes and LDH tended to increase with the duration of cement exposure. It was confirmed that the up-regulated proteins were identified as involved in glycolysis and energy metabolism and down-regulated proteins were myofibrillar proteins which were involved in muscle contraction by the cement exposure to the integument. With these results, dissolved cement powder was thought to be a big threat to the survival of the fish because it causes the morphological changes and weakens the physiological activity in C. auratus tissues.