• Journal of fish pathology
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• v.37 no.1
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• pp.17-23
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• 2024

Spring viremia of carp virus (SVCV) poses a significant threat to numerous cyprinid fish species, particularly the common carp (Cyprinus carpio), often resulting in substantial mortalities. This study explores the potential use of a chimeric recombinant snakehead rhabdovirus carrying the SVCV G gene (rSHRV-Gsvcv) as a live vaccine against SVCV infection. Through virulence testing in zebrafish at different temperatures (15 ℃ and 20 ℃), no mortality was observed in groups infected with either rSHRV-wild or chimeric rSHRV-Gsvcv at both temperatures, whereas 100% mortality occurred in fish infected with wild-type SVCV. Subsequently, as no mortality was observed by rSHRV-Gsvcv, three independent experiments were conducted to determine the possible usage of chimeric rSHRV-Gsvcv as a vaccine candidate against SVCV infection. Fish were immunized with either rSHRV-Gsvcv or rSHRV-wild, and their survival rates against the SVCV challenge were compared with a control group injected with buffer alone at four weeks post-immunization. The results showed that chimeric rSHRV-Gsvcv induced significantly higher fish survival rates compared to rSHRV-wild and the control groups. These findings suggest that genetically engineered chimeric rSHRV-Gsvcv holds the potential for a prophylactic measure to protect fish against SVCV infection.

• Korean Journal of Environmental Biology
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• v.18 no.2
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• pp.205-213
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• 2000

Laboratory studies have revealed that naturally transformable bacteria develop competence under in situ conditions. Thus, the occurrence of competent bacteria in the environment can be considered as a certainty The persistence of free DNA in natural habitats is influenced by nucleolytic degradation and protection from degradation by adsorption to minerals. Although DNA seeded into natural environment was hydrolysed at substantial rates, but was still detectable at low levels after even several weeks. Compared to the number of laboratory based studies, only a few data have been published dealing with transformation of bacteria in the field. Recently, the potential transfer of recombinant DNA (rDNA) from deliberately or accidentally released bacteria to indigenous microbes has raised biosafety issues, since the persistence of rDNA becomes independent of the survival of its original host and leads to unpredictable, long-term ecological effects. The aim of the present review is to summarise recent literature on horizontal gene transfer (HGT) by transformation among bacteria in both soil and aquatic habitat and special emphasis is placed on recent reports which have addressed HGT among bacteria in the field. [Transformation, Horizontal gene transfer (HGT), recombinant DNA (rDNA), Genetically modified microorganisms (GMMs), Biosafety]

• KSBB Journal
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• v.11 no.4
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• pp.445-452
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• 1996

To investigate the promoter effect on heterologous gene expression in S. cerevisiae, the recombinant plasmids pYI11, pYI12, pYI10-2, and pYIGP were constructed to contain the inulinase gene (INUI) as a reporter under the control of GAL10, GAL7, GAL1, and GAP promoters, respectively. When the yeasts transformants were cultivated on galactose-containing rich media, the cell growth reached to 36-39 OD600 at 72 hours of cultivation. The specific growth rates of the cells harboring the four different plasmids decreased similarly : they dropped from $0.24 h^{-1}$ during the glucose-consuming period to 0.04 -$0.10 h^{-1}$ during the galactose-consuming period (gene expression phase for GAL promoter system). After the depletion of glucose, the expression of inulinase gene was started and reached to maximal levels of 4.3(GAL1 promoter), 4.0(GAL10 promoter), 3.8(GAL7 promoter), and 1.6(GAP promoter) unit/mL at 72 hours of cultivation. Based on the maximal expression level and activity staining on the plate, the promoter strength was in the order of GAL1, GAL10, GAL7 and GAP promoter. While the GAL-promoter systems showed a high plasmid stabilities of more than 78%, the GAP-promoter plasmid revealed a lower plasmid stability of 55%. Most of inulinase activity (98%) was found in the extracellular medium, indicating that the secretion efficiency of inulinase is independent on the type of promoter.

• Journal of Life Science
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• v.17 no.5 s.85
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• pp.599-605
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• 2007

Seed and pod numbers are the main yield components in soybean. Selection for increased yield potential is main goal of plant breeding. The objective of this study was to identify quantitative trait loci(QTLs) that control pod number per plant, seed number per plant and pod in soybean. The 117 $F_{2:10}$ recombinant inbred lines(RILs) developed from a cross of 'Keunolkong' and 'Shinpaldalkong' were used. Two independent QTLs for pod number per plant were identified from linkage group(LG) F and L. Two QTLs for seed number per plant were located on LG F and L. Seed number per pod was related with three QTLs located on LG D1a, D1b and F. Pod and seed number per plant have two common QTLs on LG F and L.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 2000.04a
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• pp.178-181
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• 2000

In order to develop the novel gene expression system, we introduced new control elements which could influence the foreign gene expression in animal cells. When the foreign genes are introduced into the genome of higher eukaryotic cells, the expressions from these integrated genes are often low and can vary greatly depending on the positions of the integration sites due to the complex nature of the chromatin structures (1). First we screened the various DNA sequence elements which can function as an insulator of gene expression from these position effects and can cooperate with the SV40 enhancer/promoter. Among the several DNA elements from the various sources, we identified the particular DNA element which confers the increased frequency of the positive colonies, assayed by the reporter gene from stable selections indicating significantly reduced position effects. This element also showed the several fold-increased expression level as well as the copy-number dependent expression with host cell specificity. Second we modified the transcription termination element where we introduced the specific terminator in combination with SV40 polyA signal. This modified terminator showed the increased efficiency and the level of the gene expression. By combining these two elements, we made the animal cell expression system and tested successfully for the recombinant protein productions of TGF ${\beta}$-soluble receptor, Antithrombin III, and single chain Pro-Urokinase. [Supported by grants from MOCIE]

• Journal of Microbiology and Biotechnology
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• v.17 no.11
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• pp.1868-1874
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• 2007

We have developed a robotic system for an automated parallel cell cultivation process that enables screening of induction parameters for the soluble expression of recombinant protein. The system is designed for parallelized and simultaneous cultivation of up to 24 different types of cells or a single type of cell at 24 different conditions. Twenty-four culture vessels of about 200 ml are arranged in four columns${\times}$six rows. The system is equipped with four independent thermostated waterbaths, each of which accommodates six culture vessels. A two-channel liquid handler is attached in order to distribute medium from the reservoir to the culture vessels, to transfer seed or other reagents, and to take an aliquot from the growing cells. Cells in each vessel are agitated and aerated by sparging filtered air. We tested the system by growing Escherichia coli BL21(DE3) cells harboring a plasmid for a model protein, and used it in optimizing protein expression conditions by varying the induction temperature and the inducer concentration. The results revealed the usefulness of our custom-made cell cultivation robot in screening optimal conditions for the expression of soluble proteins.

• Journal of Crop Science and Biotechnology
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• v.11 no.4
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• pp.243-248
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• 2008

The bean bug Riptortus clavatus Thunberg (Heteroptera: Alydidae) is an important pest, causing serious yield loss in soybean. But the information on mechanism of resistance to R. clavatus is limited. The objective of this study was to identify QTLs for R. clavatus resistance using simple sequence repeat (SSR) markers in a soybean population of recombinant inbred lines (RILs) developed from the cross PI 171451 ${\times}$ Hwaeomputkong. A genetic map from this population was constructed with a total of 136 SSR markers covering 1073.9 cM on 20 linkage groups (LGs). With 126 $F_5$ RILs, two independent QTLs for resistance to R. clavatus were mapped on LGs B1 and C2. The amount of phenotypic variation explained by these QTLs ranged from 12 to 16%. PI 171451 showed an escape response to R. clavatus. Under feeding conditions, 14.4% of RILs showed greater resistance to R. clavatus than the resistant parent. The resistance to R. clavatus in soybean from PI 171451 was incomplete and quantitatively inherited and the QTLs for resistance to R. clavatus detected in the RIL population were not significantly affected by epistatic interactions.

• KSBB Journal
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• v.17 no.1
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• pp.20-25
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• 2002

We performed a basic experiment for rapid, on-line, real-time measurement of HBsAg by using a surface plasmon resonance biosensor to quantify the recognition and interaction of biomolecules. We immobilized the anti-HBsAg polyclonal antibody to the dextran layer on a CM5 chip surface which was pre-activated by N-hydroxysuccinimide for amine coupling. The binding of the HBsAg to the immobilized antibody was measured by the mass increase detected by the change in the SPR signal. The binding characteristics between HBsAg and its antibody followed typical monolayer adsorption isotherm. When the entire immobilized antibody was interacted, there was no additional, non-specific binding observed, which suggested the biointeraction was very specific as expected and independent of the ligand density. No significant steric hindrance was observed at 17.6 nm/$mm^2$ immobilization density. The relationship between the HBsAg concentration in the sample solution and the antigen bound to the chip surface was linear up to ca. $40\mu\textrm{g}$/mL, which is much wider than that of the ELISA method. It appeared the antigen-antibody binding was increased as the immobilized ligand density increased, but verification is warranted. This study showed the potential of this biosensor-based method as a rapid, simple, multi-sample, on-line assay. Once properly validated, it can serve as a more powerful method for HBsAg quantification replacing the current ELISA method.

• Experimental and Molecular Medicine
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• v.50 no.12
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• pp.8.1-8.12
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• 2018

FAM46B is a member of the family with sequence similarity 46. Little is known about the expression and functional role (s) of FAM46B in prostate cancer (PC). In this study, the expression of FAM46B expression in The Cancer Genome Atlas, GSE55945, and an independent hospital database was measured by bioinformatics and real-time PCR analysis. After PC cells were transfected with siRNA or a recombinant vector in the absence or presence of a ${\beta}$-catenin signaling inhibitor (XAV-939), the expression levels of FAM46B, C-myc, Cyclin D1, and ${\beta}$-catenin were measured by western blot and realtime PCR. Cell cycle progression and cell proliferation were measured by flow cytometry and the CCK-8 assay. The effects of FAM46B on tumor growth and protein expression in nude mice with PC tumor xenografts were also measured. Our results showed that FAM46B was downregulated but that ${\beta}$-catenin was upregulated in patients with PC. FAM46B silencing promoted cell proliferation and cell cycle progression in PC, which were abrogated by XAV-939. Moreover, FAM46B overexpression inhibited PC cell cycle progression and cell proliferation in vitro and tumor growth in vivo. FAM46B silencing promoted ${\beta}$-catenin protein expression through the inhibition of ${\beta}$-catenin ubiquitination. Our data clearly show that FAM46B inhibits cell proliferation and cell cycle progression in PC through ubiquitination of ${\beta}$-catenin.

• Journal of Korean Neurosurgical Society
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• v.62 no.2
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• pp.201-208
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• 2019

Objective : In patients with internal carotid artery (ICA) occlusion, collateral middle cerebral artery (MCA) flow has a protective role against ischemia. However, some of these patients may experience initial major neurological deficits and major worsening on following days. Thus, we investigated the safety and efficacy of endovascular treatment (EVT) for ICA occlusion with collateral MCA flow by comparing clinical outcomes of medical treatment versus EVT. Methods : The inclusion criteria were as follows : 1) acute ischemic stroke with ICA occlusion and presence of collateral MCA flow on transfemoral cerebral angiography (TFCA) and 2) hospital arrival within 12 hours from symptom onset. The treatment strategy was made by the attending physician based on the patient's clinical status and results of TFCA. Results : Eighty-one patients were included (30 medical treatment, 51 EVT). The EVT group revealed a high incidence of intracranial ICA occlusion, longer ipsilesional MCA contrast filling time, and a similar rate of favorable clinical outcome despite a higher mean baseline the National Institutes of Health Stroke Scale (NIHSS) score. By binary logistic regression analysis, intravenous recombinant tissue plasminogen activator and EVT were independent predictors of favorable clinical outcome. In subgroup analysis based on stroke etiology, the non-atherosclerotic group showed a higher baseline NIHSS score, higher incidence of EVT, and a higher rate of distal embolization during EVT in comparison with the atherosclerotic group. Conclusion : In patients with ICA occlusion and collateral MCA flow, decisions regarding treatment strategy based on TFCA can help achieve favorable clinical outcomes. EVT strategy with respect to etiology of ICA occlusion might help achieve better angiographic outcomes.