• Development and Reproduction
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• v.19 no.3
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• pp.135-144
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• 2015

Heat shock protein (HSP) 70, the highly conserved stress protein families, plays important roles in protecting cells against heat and other stresses in most animal species. In the present study, we identified and characterized four Hsp70 (RuHSP4, RuHSC70, RuHSP12A, RuGRP78) family proteins based on the expressed sequence tag (EST) analysis of the Korean rose bitterling R. uyekii cDNA library. The deduced RuHSP70 family has high amino acid identities of 72-99% with those of other species. Phylogenetic analysis revealed that RuHsp70 family clustered with fish groups (HSP4, HSC70, HSP12A, GRP78) proteins. Quantitative RT-PCR analysis showed the specific expression patterns of RuHsp70 family members in the early developmental stages and several tissues in Korean rose bitterling. The expression of 4 groups of Hsp70 family was detected in all tested tissue. Particularly, Hsp70 family of Korean rose bitterling is highly expressed in hepatopancreas and sexual gonad (testis and ovary). The expression of Hsp70 family was differentially regulated in accordance with early development stage of Rhodeus uyekii.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.15 no.8
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• pp.5412-5418
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• 2014

The red seabream iridovirus (RSIV), which belongs to the iridoviridae, causes infectious fish diseases in many Asian countries, leading to considerable economic losses to the aquaculture industry. Using the yeast surface display (YSD) technique, a new experimental system was recently developed for the detection and identification of a variety of marine viruses. In this study, a coat protein gene of RSIV was synthesized based on the nucleotide sequence database and subcloned into the yeast expression vector, pCTCON2. The expression of viral coat proteins in the yeast strain, EBY100, was detected by flow cytometry and Western blot analysis. Finally, they were isolated from the yeast surface through a treatment with ${\beta}$-mercaptoethanol. The data suggests that the YSD system can be a useful method for acquiring coating proteins of marine viruses.

• The Korean Journal of Zoology
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• v.38 no.4
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• pp.557-564
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• 1995

The 90 kDa-heat shock protein (HSP90) is one of the major ubiquitous heat shock proteins induced by a vadety of ceilular stresses. HSP90 Is constitutively synthesized even under nonstressed condidons and found In association with several regulatory and structural proteins such as protein kinases and steroid hormone receptors. In the present study, to facilitate its biochemical characterization, HSP90 was pudfied from chick muscle by sequential column chromatography steps including DEAE- cellulose, hydroxyapatite, and Sephacryl S-300 gel filtration and monoclonal antillodies specific to HSP90 were produced by the inurine hybridomal technique. We report the production of 4 posItive hybridoma clones, named as A204, C112, C302 and A204, C112, C302. Among these MoAbs, Cl 12 strongly reconnized chick HSP90 in Western blot and native immunoprocipitation. In addition, C112 showed the crossreactivitles against HSP90 from human, rabbit, mouse, fish and chick but not from Drosophila and E. coil.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2003.10a
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• pp.111-111
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• 2003

Telomeres are the end of chromosomes and consist of a tandem repeat sequence of (TTAGGG)n and associated proteins. Telomerase is a ribonucleoprotein which act as a template for the synthesis of telomeric DNA. Telomeres are essential for chromosome stability and are related with cell senescence, apoptosis and cancer. Even though telomeres and telomerase have been studied extensively, very little is known about telomere dynamics in embryonic cells. This study was carried out to analyze the telomeres distribution and telomerase activity of chicken cells during embryonic and developmental stages. The target cells for analysing were sperms, ovulated ova, early embryonic cells and the cells from brain, heart, liver, kidney and germinal tissue in fetus. Telomeres distribution on target cells was analyzed by Q-FISH (Quantitation-Fluorescence in situ Hybridization) techniques using a chicken telomere repeat probe. Telomerase activity was performed by TRAP assay (Telomeric repeat Amplification Protocol) with target DNA. In results, the telomeres of chicken were found at the ends of all chromosomes. In addition, chicken had interstitial telomeres on chromosomes 1, 2 and 3. Telomerase activity was highly detectable in early embryonic cells, germinal tissues and kidney cells. Whereas telomerase activity was gradually down-regulated when the organs, including brain, heart, and liver, were developed from embryos. In the distribution of telomeric DNA on the embryonic and developmental stages, most of the cells was gradually decreased in telomere quantity during ontogenesis.

• Development and Reproduction
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• v.14 no.2
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• pp.107-113
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• 2010

The olive flounder (Paralichthys olivaceus) is one of the most widely cultured flatfish in Korea and Japan. During development, in a process known as metamorphosis, this fish reorients itself to lie on one side, the body flattens, and the eye migrates to the other side of the body. However, few studies have focused on molecule regulation mechanism of eye development in olive flounder. To reveal the molecular mechanism of eye development, we performed the studies on identification of genes expressed in the eye of olive flounder using EST and RT-PCR strategy. A total of 270 ESTs were sequenced, and 178 (65.9%) clones were identified as known genes and 92 (34.1%) as unknown genes. Among the 178 EST clones, 29 (16.3%) clones were representing 9 unique genes identified as homologous to the previously reported olive flounder ESTs, 131 (73.6%) clones representing 107 unique genes were identified as orthologs of known genes from other organisms. We also identified a kind of eye development associated proteins, indicating EST as a powerful method for identifying eye development-related genes of fish as well as identifying novel genes. Further functional studies on these genes will provide more information on molecule regulation mechanism of eye development in olive flounder.

• Korean Journal of Veterinary Research
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• v.38 no.2
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• pp.371-378
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• 1998

Five strains of gram-negative and yellow-pigmented bacteria were recently isolated from diseased freshwater fishes in Korea. All isolates were confirmed as a known fish pathogen of columnaris disease, Cytophaga columnaris based on their colonial and cellular morphology, and on physiological, biochemical and antigenic characteristics. Although the isolates were from different fish species, their characteristics of them were very similar to those of the reference strains of C columnaris (NCMB $2248^T$ and EK 28). Also, profiles of OMPs of Korean isolates were similar to those of the reference strain, C columnaris NCMB $2248^T$ when analyzed by SDS-PAGE.

• Journal of Marine Bioscience and Biotechnology
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• v.2 no.1
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• pp.68-72
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• 2007

Flounder is one of the major aquacultured fish and of an economically important item in Korean fisheries. Recently, there are trends of research worldwide that aim to analyze and characterize a whole genome or a whole proteome of interesting species. The data are utilized for the understanding and development of preventive and curative technologies for the serious diseases. However, there are very limited information of proteome for marine organisms, we optimized first the conditions for two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) with serum form a marine fish, flounder (Paralichthys olivaceus). A pre-treatment of serum and an optimization of protein concentration analyzed were surveyed for enhancing the separation for proteins. A statistical analysis was performed on the overall 1,820 protein spots to overcome the variability among individual fishes.

• Fisheries and Aquatic Sciences
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• v.23 no.8
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• pp.22.1-22.8
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• 2020

Background: High demand and low supply of fishmeal due to overexploitation of fisheries resources have resulted in a dramatic increase in the price of this ingredient. Olive flounder (Paralichthys olivaceus) commercial feed contains approximately 60% fishmeal and limited success has been achieved in identifying sustainable alternative protein sources for this species. Methods: An on-farm feeding trial was conducted to compare a basal diet containing 65% as the control (CONT) with two experimental diets replacing 10% of fishmeal by animal protein (AP₁₀) or 20% of fishmeal by animal and plant protein (APP₂₀). Sub-adult olive flounder averaging 327 ± 9.3 g (mean±SD) were fed one of the three diets in triplicate groups for 16 weeks. Results: Weight gain, specific growth rate, feed efficiency, protein efficiency ratio, and survival were not significantly different among fish fed all the experimental diets (P > 0.05). Also, non-specific immune responses (superoxide dismutase and lysozyme activity), serum biochemical parameters, and intestinal villi length were not significantly different among fish fed all the experimental diets (P > 0.05). Conclusions: Therefore, based on growth performance, non-specific immune responses, serum biochemical parameters, and intestinal histology, dietary animal and plant protein mixtures could replace up to 20% of fishmeal in the diet of sub-adult olive flounder.

• Journal of Animal Science and Technology
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• v.47 no.2
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• pp.187-194
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• 2005

Telomeres locate at the end of chromosomes and consist of a tandem repeat sequence of $(TIAGGG)^{n}$ and associated proteins. Telomerase is a ribonucleoprotein which act as a template for the synthesis of telomeric DNA. Telomeres are essential for chromosome stability and are related with cell senescence, apoptosis and cancer. This study was carried out to analyze the amount of telomeres and telomerase activity of chicken cells during embryonic and developmental stages. The whole embryos and prenatal tissues such as brain, heart, liver, kidney and testis at different developmental stages were obtained from Korean Native Chicken. The amount of telomeres on embryonic cells was analyzed by quantitative fluorescence in situ hybridization (Q-FISH) techniques using the chicken telomeric DNA probe. Telomerase activity was measured by telomeric repeat amplification protocol (TRAP) assay. Results indicated that the amounts of telomeric DNA on the most embryonic cells were gradually decreased during ontogenesis. Furthermore, the quantity of telomeres was quite different among embryonic tissues according to developmental origin. The relative amount of telomeres has more in regenerative cells such as embryonic disc and testicular cells than in non-regenerative cells such as liver, brain, heart and kidney cells. Telomerase activity was also highly detectable in most chicken cells at early embryonic stages. After 9 days of incubation, however, the telomerase activitie W

• Journal of fish pathology
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• v.21 no.1
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• pp.1-11
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• 2008

We report the existence of new type of phosphatidylcholine-hydrolyzing phospholipase D (PLD), which has been characterized and partially purified in the scuticociliate, Uronema marinum. The enzyme from partial purification showed that it was existed in membrane fraction and was a neutral PLD, which catalyzed both transphosphatidylation and hydrolysis reaction. The activity of partially purified membrane-bound PLD was also found to be optimal at pH 7.0-7.5 for 2 hours at 37℃ and depended strictly on the presence of Ca2+ (2.5 mM) and Mg2+ (1.6 mM). Immunoblot analysis indicated that the enzyme was distinct from hPLD1 (human PLD1) and hPLD2 (human PLD2) because it was not recognized by a polyclonal antibody raised to the 12 terminal amino acid of these enzymes. We also found that the membrane-bound PLD is a PIP2-dependent PLD and that GTP-binding proteins are not implicated in the regulation of this enzyme: This enzyme activity is markedly stimulated by phosphatidylinositol 4,5-bisphosphate (PIP2) but not by the small G-protein Arf and GTPrS. In addition, this enzyme was capable of hydrolyzing phosphatidylcholine (PC) but not phosphatidylethanolamine (PE), implying that PC was a preferred substrate.