• BMB Reports
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• v.40 no.2
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• pp.286-289
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• 2007

Despite considerable interest in the biologic processes of regeneration and stem cell activation, little is known about the genes involved in these transformative events. In a Hydra littoralis model of regeneration, we employed a rapid shotgun suppression subtractive hybridization strategy to identify genes that are uniquely expressed in regenerating tissue. With an adaptor-PCR based technique, 16 candidate transcripts were identified, 15 were confirmed unique to mRNA isolated from hydra undergoing regeneration. Of these, 6 were undescribed in GenBank and allied expressed sequence tag (EST) databases (GenBank + EMBL + DDBJ + PDB and the Hydra EST database). BLAST analysis of these sequences identified remarkably similar sequences in anonymous ESTs found in a wide variety of animal species.

• Biotechnology and Bioprocess Engineering:BBE
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• v.8 no.4
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• pp.263-267
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• 2003

Magnetic nanoparticles prepared from an alkaline solution of divalent and trivalent iron ions could covalently bind protein via the activation of Nethyl-N-(3-dimethylaminopropyl) carbodiimide (EDC). Trypsin and avidin were taken as the model proteins for the formation of protein-nanoparticle conjugates. The immobilized yield of protein increased with molar ratio of EDC/nanoparticie. Higher concentrations of added protein could yield higher immobilized protein densities on the particles. In contrast to EDC, the yields of protein immobilization via the a ctivation of cyanamide were relatively lower. Nanoparticles bound with avidin could attach a single-stranded DNA through the avidin-biotin interaction and hybridize with a DNA probe. The DNA hybridization was confirmed by fluorescence microscopy observations. Immobilized DNA on nanoparticles by this technique may have widespread applicability to the detection of specific nucleic acid sequence and targeting of DNA to particular cells.

• Proceedings of the Korean Society of Veterinary Pathology Conference
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• 2003.10a
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• pp.28-28
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• 2003

Hepatitis E virus (HEV), previously referred to as enterically transmitted non-A, non-B hepatitis, is responsible for sporadic infections as well as large epidemics of acute viral hepatitis in developing countries. The disease generally affects young adults and reportedly has a mortality rate of up to 20％ in infected pregnant women. HEV was once considered to be a member of the family Caliciviridae, but the unique genomic organization of HEV has led to the removal of HEV from the family and it was provisionally classified in an unassigned family of HEV-like viruses. In situ hybridization provides any cellular detail and histological architecture.[1] However, use of in situ hybridization is largely restricted to the laboratories because this technique is the greater technical complexity and expense compared with immunohistochemistry. Therefore, the objective of this study is to develop the immunohistochemistry for the detection of swine HEV from formalin-fixed, paraffin-embedded hepatic tissues. (omitted)

• Korean Journal of Microbiology
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• v.26 no.3
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• pp.155-161
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• 1988

Nucleotide sequence homology between bovine, simian, and porcine rotavirus was determined by the RNA:RNA hybridization technique. Single stranded RNA, prepared in vitro with EDTA activated endogeneous viral RNA polymerase, was hhbridized with tritium labeled bovine rotavirus genomic RNA. The heteroduplex RNA was treated with single stranded RNA specific ribonucleases and the RNase resistant hybrid RNA was precipitated, and collected by filtration on a filter paper. Seventy four percent RNA sequence homology between bovine and simian rotavirus and 8 percent RNA sequence homology between bovine and porcine rotavirus was confirmed by hybridization between tritium labeled single stranded RNA and viral genomic RNA.

• Korean Journal of Microbiology
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• v.25 no.3
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• pp.173-179
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• 1987

Ti plasmids were isolated from three strains of Agrobacterium tumefaciens in Korea and their types and molecular weights were determined. All of these are octopine-type and their molecular weights are 44Kb (pTi 12), 180Kb (pTi 14) and 172Kb (pti 49), respectively. In order to construct physical map of pTi 12, pTi 12 was digested with restriction endonucleases Sma I and Hind III. Sma I degestion of pTi 12 produce 8 fragments and Hind III produced 10 fragments. Physical arrangements of these fragments was determined by Southern hybridization techniques.

• Korean Journal of Veterinary Research
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• v.36 no.4
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• pp.845-858
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• 1996

The purpose of this study was to establish early diagnostic methods for the detection of Aujeszky's disease viral antigens and nucleic acid in nasal cells, and buffy coats from experimentally infected living pigs by a combination of immunocytochemistry, in situ hybridization with digoxigenin(DIG)-labled probe and electron microscopy. Forty days old piglets were inoculated intranasally with $10^{7.0}TCID_{50}$ of Aujeszky's disease virus (ADV, NYJ-1-87 strain). The viral antigens and nucleic acid of ADV were detected in nasal cells, and buffy coat for 20 days after inoculation by immunocytochemistry, in situ hybridization with DIG-labeled probe and electron microscopical method. The results were compared with conventional methods such as a porcine Aujeszky's disease serodiagnostic(PAD) kit, neutralization test(NT) and virus isolation. 1. The viral antigens, nucleic acids and capsids of ADV were detected in nasal cells, buffy coats from 3 days to 20 days after inoculation by immunocytochemistry, in situ hybridization with DIG-labeled probe and electron microscopy, respectively. 2. When viral antigens were detected by the immunocytochemical technique, a diffuse brown deposit was observed in the nucleus and cytoplasm of nasal cells, buffy coats and PK-15 cells under a microscope. 3. DIG-labeled DNA probe was prepared by amplification of conserved sequence of recombinant ADV-gp50 clone with polymerase chain reacction. When ADV-DNA was detected by ISH with DIG-labeled probe, purplish blue pigmentation were observed in the nuclei and cytoplasms of ADV-infected cells under a microscope. Positive signals were observed in nasal cells and in the buffy coat and PK-15 cells at the first day after inoculation. 4. Where ADV-capsids were detected by transmission electron microscopical method, aggregation of capsids was observed in the nuclei and cytoplasms of nasal cells, buffy coats and PK-15 cells. The results suggested that these methods were considered as the highly sensitive and reliable tools for rapid and confirmative diagnosis of Aujeszky's disease in living pigs.

• Proceedings of the Materials Research Society of Korea Conference
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• 2009.11a
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• pp.6.2-6.2
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• 2009

Nanostructured materials arecurrently receiving much attention because of their unique structural andphysical properties. Research has been stimulated by the envisagedapplications for this new class of materials in electronics, optics, catalysisand magnetic storage since the properties derived from nanometer-scalematerials are not present in either isolated molecules or micrometer-scalesolids. This study presents the experimental results derived fromthe various functional materials processed in nano-scale using pulsed laserablation, since those materials exhibit new physical phenomena caused by thereduction dimensionality. This presentation consists of three mainparts to consider in pulsed laser ablation (PLA) technique; first nanocrystallinefilms, second, nanocolloidal particles in liquid, and third, nanocoating fororganic/inorganic hybridization. Firstly, nanocrystalline films weresynthesized by pulsed laser deposition at various Ar gas pressures withoutsubstrate heating and/or post annealing treatments. From the controlof processng parameters, nanocystalline films of complex oxides and non-oxidematerials have been successfully fabricated. The excellentcapability of pulsed laser ablation for reactive deposition and its ability totransfer the original stoichiometry of the bulk target to the deposited filmsmakes it suitable for the fabrication of various functionalmaterials. Then, pulsed laser ablation in liquid has attracted muchattention as a new technique to prepare nanocolloidal particles. Inthis work, we represent a novel synthetic approach to directly producehighly-dispersed fluorescent colloidal nanoparticles using the PLA from ceramicbulk target in liquid phase without any surfactant. Furthermore, novel methodbased on simultaneous motion tracking of several individual nanoparticles isproposed for the convenient determination of nanoparticle sizedistributions. Finally, we report that the GaAs nanocrystals issynthesized successfully on the surface of PMMA (polymethylmethacrylate)microspheres by modified PLD technique using a particle fluidizationunit. The characteristics of the laser deposited GaAs nanocrytalswere then investigated. It should be noted that this is the first successfultrial to apply the PLD process nanocrystals on spherical polymermatrices. The present process is found to be a promising method fororganic/inorganic hybridization.

• Journal of Korean Society of Environmental Engineers
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• v.22 no.5
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• pp.939-947
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• 2000

Enhanced biological phosphorus removal (EBPR) was performed in an anaerobic/aerobic sequencing batch reactor (SBR). Influent was a synthetic wastewater based on acetate as a carbon source. The sludge age and hydraulic retention time were kept at 10 days and 16 hrs, respectively, Phosphate release during the anaerobic period and phosphate uptake in aerobic period were increased gradually with time. and after about 200 days, steady-state operation could be achieved with complete removal of influent phosphate. Number distribution of microbial community in the sludge performing EBPR was investigated during the steady state operation. 17 rRNA targeted oligonucleotide probes were designed and slot hybridization technique was used to determine the number distribution of each microorganism. In the acetate fed SBR, rRNA belonging to the beta subclass of proteobacteria was the most dominant in total rRNA and rRNA matching to CTE probe was the second, rRNAs of Acinetobacter, Aeromonas and Pseudomonas, which are usually thought as phosphorus accumulating organisms in EBPR processes, constituted less than 10% of total rRNA. From this community analysis, it was inferred that microorganisms belong to the beta subclass of proteobacteia (BET) and CTE such as Rhodocyclus group were important in biological phosphorus removal. Therefore, the role of Acinetobacter, Aeromonas and Pseudomonas in the EBPR might have been overestimated.

• Korean Journal of Microbiology
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• v.34 no.3
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• pp.169-174
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• 1998

To define the structure and diversity of bacterial communities in the aqutic ecosystem, Lake Soyang, the largest artificial reservoir in Korea, a new method, fluorescent in situ hybridization was applied. This technique relies on the specific hybridization of the nucleic acid probes to the naturally amplified intracellular rRNA. By this method, the bacterial community composition of Lake Soyang and bacterial numbers belong to eubacteria, proteobacteria and Cytophaga-Flavobacterium group were estimated. Total bacterial numbers ranged from $0.3{\times}10^6{\sim}2.0{\times}10^6cells{\cdot}ml^{-1}$, and vertical profile of total bacteria showed the peak at 2 and 5 m depths. The ratio of eubacteria to total bacteria were 22~100% and varied with depth and season. The percentage of Proteobacteria ${\alpha}$-group ranged 2.6~66.7%, ${\beta}$-group 4.5~53.5%, ${\gamma}$-group 4.6~76.7% and Cytophaga-Flavobacterium group 2.1~35.9%. Also, bacteria] community had spatial and temporal characteristics. The dominant groups were ${\beta}$-group in winter, ${\gamma}$-group in spring and early summer and ${\alpha}$-group in summer.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.20 no.8
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• pp.627-633
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• 2019

Rapid and accurate identification of marine microalgae causing harmful algal blooms (HABs) is a crucial tool for predicting and managing HABs. We previously developed a nuclease protection assay sandwich hybridization (NPA-SH) method for the in situ detection of blooming microalgae Cochlodinium polykrikoides. In this study, we improved the applicability of the NPA-SH method for the detection of C. polykrikoides by simplifying the reaction step. For this purpose, invertase (INV) was conjugated to the signal probe instead of using fluorescence, and sucrose was used as a reactant to induce a color reaction. The INV-signal probe conjugation was confirmed by SDS-PAGE and epifluoromicroscopy. The treatment time and appropriate amounts of the probe and sucrose that optimized the reaction were determined. As a result, the developed INV-SH reduced the treatment time in the field compared with NPA-SH, and also enabled the use of a relatively small volume and low-priced personal glucose meter, as well as an absorbance meter. INV-SH is the first C. polykrikoides species identification technology to which INV has been applied and could be an improved field technique.