• Title/Summary/Keyword: 서열화

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Replication and Pathogenesis of Plaque Morphology Mutants Derived from Vero Cells with Japanese Encephalitis Virus Persistency (지속감염세포에서 분리된 일본뇌염바이러스 Plaque Morphology Mutants의 복제 및 감염특성)

  • 윤성욱;정용석
    • Korean Journal of Microbiology
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    • v.38 no.3
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    • pp.221-229
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    • 2002
  • Japanese encephalitis virus (JEV) persistence was established and maintained in Vero cell culture for over 1 year. Eleven clones of plaque morphology mutant JEV, with large and small plaque sizes, were obtained from the cell culture supernatant. Genomic RNA replication efficiency of the mutants in naive Vero cell appeared to correspond to their different plaque sizes. No significant changes in envelop protein ORF or in non-coding regions at both ends of the RNA genome suggested that there could be an unidentified factor(s) playing role in JEV attenuation. Unlike to the replication of wild-type JEV, the mutants did not induce severe degree of cytopathic effect in Vero cells upon infection. While obvious decrease of Bcl-2 and its mRNA expression and sharp increase of p53 in naive Vero cells infected with either wild-type JEV or the large plaque-forming mutant, those changes were not observed with the small plaque-forming one. Together with these observation, internucleosomal DNA fragmentation and chromosomal DNA profile in the Vero cells infected with the mutants suggest that an overall changes in cytopathic effect in the plaque morphology mutants-infected cells should be primarily due to the reduced genomic RNA replication and the compromised degree of p53-independent apoptosis by the virus infection at least in part.

Isolation and Characteristics of Bacteria Showing Biocontrol and Biofertilizing Activities (생물방제 및 생물비료 활성을 가지는 세균의 분리 및 특성)

  • Jung, Ho-Il;Kim, Keun-Ki;Park, Hyean-Cheal;Lee, Sang-Mong;Kim, Yong-Gyun;Kim, Hong-Sung;Lee, Cnung-Yeol;Son, Hong-Joo
    • Journal of Life Science
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    • v.17 no.12
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    • pp.1682-1688
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    • 2007
  • To develop multifunctional microbial inoculant, microorganisms with antagonistic activity and biofertilizing activity were screened. Pantoea agglomerans and Bacillus megaterium from our laboratory culture collection, and strain MF12 from soil near poultry farm in Miryang were selected. On the basis of morphological, physiological studies and 16S rDNA sequence analysis, isolate MF12 was identified as the Bacillus pumilis. Three strains were studied for insoluble phosphate solubilization, indole-3-acetic acid (IAA) and siderophore production, ammonification ability, hydrolytic enzyme production and antifungal activity against phytopathogenic fungi. P. agglomerans did not produce any visible clear zone on agar plate containing 0.5% $Ca_3(PO_4){_2}$ as a sole phosphorus source. However, this strain could solubilize insoluble phosphate in liquid medium. All strains produced IAA ranged from $3{\sim}639{\mu}g/ml$ depending on culture time and had ammonification ability. Among three strains, only P. agglomerans produced siderophore. P. agglomerans produced pectinase and lipase, B. megaterium produced amylase, protease and lipase while B. pumilis produced protease and lipase. P. agglomerans showed antifungal activities against phytopathogenic fungi, Fusarium oxysporum and Colletotrichum gloeosporioides. B. pumilis showed antifungal activities against Botrytis cinerea, Sclerotinia sclerotiorum and Phythium ultimum.

K+ Ion-Selective PVC Membrane Electrodes with Neutral Carriers (중성운반체를 이용한 K+ 이온선택성 PVC막 전극)

  • Kim, Yong-Ryul;Cho, Kyoung-Sub;Kang, An-Soo
    • Applied Chemistry for Engineering
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    • v.9 no.5
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    • pp.734-741
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    • 1998
  • Electrode characteristics were studied in the interface between sample solutions and $K^+$ ion selective PVC membrane electrodes containing neutral carriers, dibenzo-18-crown-6(D18Cr6) and valinomycin(Val). The effect of doping of base electrolytes, the chemical structure and the content of carrier, variation of plasticizer, membrane thickness, and concentration variation of sample solution on the response characteristics of electrode such as the measured Nernstian slope, the detection limit, the linear response range, and potentiometric selectivity coefficients, were studied. In order to synthesize the membrane D18Cr6 and Val as neutral carriers were used, and complex between the carrier and $K^+$ ions were used as active materials. PVC membrane electrodes were made of plasticizers (DBP, DOS, and DBS), the base electrolyte[potassium tetraphenylborate(KTPB)], and solvent(THF). The chemical structure of carrier D18Cr6 was best for electrode and ideal electrode characteristics were appeared especially in case of doping of TPB. The optimum carrier content was about 3.23 wt % in case of D18Cr6 and Val. DBP was best as a plasticizer. As membrane thickness decreased the electrode characteristics was improved. But its characteristics were lowered below the optimum membrane thickness because of the elution of carrier, deterioration of membrane strength, etc. In the case of D18Cr6, the selectivity coefficients by the mixed solution method for the $K^+$ ion were in the order of $NH_4{^+}>Ca^{2+}>Mg^{2+}>Na^+$.

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Isolation and Characterization of a Marine Bacterium Producing Thermotolerant Agarase (내열성 한천분해효소를 생산하는 해양세균의 분리 및 특성)

  • Park Ceun-Tae;Lee Dong-Ceun;Kim Nam Young;Lee Eo-Jin;Jung Jong-Ceun;Lee Jae-Hwa;Heo Moon-Soo;Lee Jung-Hyun;Kim Sang-Jin;Lee Sang-Hyeon
    • Journal of Life Science
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    • v.15 no.6 s.73
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    • pp.884-888
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    • 2005
  • An agar-degrading bacterium was isolated from north-eastern sea of Jeju island and cultured in marine agar 2216 media. Biochemical and morphologicl characteristics and 165 rRNA gene revealed that isolated strain was member of Agarivorans genus, and named Agarivorans sp. JA-1. Agarase was produced as growth-related and expressed regardless of agar presence. Optimal pH was 8 at 50 mM Clycine-NaOH buffer, and activity was maximum at $40^{\circ}C$E Enzymatic activity was maintained over $80\%$ at $60^{\circ}C$t and $70\%$ at $80^{\circ}C$ which is thermotolerant. Hence isolated novel Agarivorans sp. JA-1 strain and its beta-agarase could be used for the production of functional oligosaccharide from agar in solution state.

Mass Mortalities of Cultured Striped Beakperch, Oplegnathus fasciatus by Iridoviral Infection (이리도 바이러스 감염에 의한 양식 돌돔, Oplegnathus fasciatus의 대량폐사)

  • Sohn, Sang-Gyu;Choi, Dong-Lim;Do, Jeung-Wan;Hwang, Jee-Youn;Park, Jeong-Woo
    • Journal of fish pathology
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    • v.13 no.2
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    • pp.121-127
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    • 2000
  • From August to October 1998, over 60% mortality of cultured striped beakperch Oplegnathus fasciatus was occurred in net cages along the southern coast of Korea. Moribund fish showed some clinical signs of lethargic behavior, dark coloration or decoloration, severe gill anemia and enlargement of spleen. Also enlarged basophilic cells showing Feulgen -positive reaction were observed in the tissue section of spleen, kidney, liver and heart of the diseased fish. GF cells inoculated with spleen homogenate of diseased fish produced cytopathic effect of enlarged and rounded cells, therefore the causative virus was isolated from diseased fish. Striped beakperch fingerlings intraperitoneally inoculated with the causative virus ($10^4TCID_{50}$/0.1 ml) revealed symptoms similar to those of naturally infected fish and died from 7 to 14 days post injection. Transmission electron microscopy revealed that the causative virus was enveloped icosahedral particle with 120~130 nm in diameter. PCR products of the expected size (500 bp) were amplified with a primer set based on the ATPase gene of RSIV(red sea bream iridovirus) using template DNAs which were extracted from the spleen of diseased fish and GF cells inoculated with the causative virus. According to the analysis of nucleotide sequence of these PCR products, the sequence from ATPase cDNA gene of the causative virus showed 95% homology with that of RSIV. These results indicate that the mass mortality in the cultured striped beakperch was caused by the infection of iridovirus similar to RSIV.

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Diversity and physiological properties of soil actinobacteria in Ulleung Island (울릉도 유래 토양 방선균의 다양성과 생리활성)

  • Yun, Bo-Ram;Roh, Su Gwon;Kim, Seung Bum
    • Korean Journal of Microbiology
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    • v.53 no.4
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    • pp.242-250
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    • 2017
  • Actinobacteria tolerating extreme conditions can be a rich source of bioactive compounds and enzymes. In this study filamentous actinobacteria were isolated from soils of Ulleung Island, and their physiological properties were examined. Soil samples were collected, serially diluted and spread on various agar media. The average viable counts of total bacteria were $1.28{\times}10^7CFU/g$ for soil sample 1 (ULS1) and $2.05{\times}10^7CFU/g$ for soil sample 2 (ULS2). As a result, 34 strains of actinobacteria were isolated and assigned to the genera Streptomyces (16 strains), Isoptericola (5 strains), Rhodococcus (4 strains), Agromyces (3 strains), Micrococcus (2 strains), Arthrobacter (1 strain), Williamsia (1 strain), Microbacterium (1 strain), and Oerskovia (1 strain) based on 16S rRNA gene sequence analysis. Enzyme activity and plant growth promoting potential were tested for representative isolates. Multiple strains of Streptomyces degraded starch, casein and Tween 80. As for plant growth promoting potential, strains of Oerskovia, Williamsia, Isoptericola, and Streptomyces solubilized phosphate, and those of Agromyces, Oerskovia, Micrococcus, Rhodococcus, Streptomyces, and Isoptericola produced 3-indole-acetic acid (IAA), respectively. Selected strains of Streptomyces exhibited strong antagonistic activity against Staphylococcus aureus and Bacillus subtilis as well as Candida albicans. This study confirms that actinobacteria from Ulleung Island can be a good source of novel bioactive compounds.

Application and Utilization of Environmental DNA Technology for Biodiversity in Water Ecosystems (수생태계 생물다양성 연구를 위한 환경유전자(environmental DNA) 기술의 적용과 활용)

  • Kwak, Ihn-Sil;Park, Young-Seuk;Chang, Kwang-Hyeon
    • Korean Journal of Ecology and Environment
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    • v.54 no.3
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    • pp.151-155
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    • 2021
  • The application of environmental DNA in the domestic ecosystem is also accelerating, but the processing and analysis of the produced data is limited, and doubts are raised about the reliability of the analyzed and produced biological taxa identification data, and the sample medium (target sample, water, air, sediment, Gastric contents, feces, etc.) and quantification and improvement of analysis methods are also needed. Therefore, in order to secure the reliability and accuracy of biodiversity research using the environmental DNA of the domestic ecosystem, it is a process of actively using the database accumulated through ecological taxonomy and undergoing verification procedures, and experts verifying the resolution of the data increased by gene sequence analysis. This is absolutely necessary. Environmental DNA research cannot be solved only by applying molecular biology technology, and interdisciplinary research cooperation such as ecology-taxa identification-genetics-informatics is important to secure the reliability of the produced data, and researchers dealing with various media can approach it together. It is an area in desperate need of an information sharing platform that can do this, and the speed of development will proceed rapidly, and the accumulated data is expected to grow as big data within a few years.

The Rapid Detection of Pathogens in Organically Grown Vegetables Using PCR-DGGE (PCR-DGGE를 이용한 유기농 채소의 유해 미생물 신속 검지)

  • Kwon, Oh Yeoun;Son, Seok Min
    • Food Engineering Progress
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    • v.15 no.4
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    • pp.370-375
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    • 2011
  • In this study the polymerase chain reaction (PCR) combined with denaturing gradient gel electrophoresis (DGGE) was evaluated as a method permitting the rapid detection of pathogens in fresh originally grown vegetables. A universal primer (341GCf/534r) was selected for its ability to amplify the V3 region of 16S-rRNA genes in their target pathogens (Salmonella typhimurium, Pseudomonas fluorescens, Bacillus cereus, Listeria monoytogenes, Staphyloocus aureus, E. coli). The 194 bp fragments in PCR were successfully duplicated as expected. The amplified fragments of the same size from six different pathogens also showed good separation upon DGGE. The detection limit of PCR-DGGE for six pathogens in fresh-cut lettuces were over $10^{5}$ CFU/g when sampled by stomaching. However, when the sampling method was changed from stomaching to shaking, the detection limit of six pathogens in organic vegetables was shown to increase by over $10^{1}$ CFU/g, but only those of B. cereus were over $10^{3}$ CFU/g. Therefore, PCR-DGGE was shown to be a reliable method for the detection of pathogens in fresh-cut vegetables.

Changes in pathogenic characters of Pseudomonas tolaasii 6264 strain by storage period (저장 기간에 따른 Pseudomonas tolaasii 6264 균주의 병원 특성 변화)

  • Yun, Yeong-Bae;Huh, Jeong-Hun;Kim, Young-Kee
    • Journal of Applied Biological Chemistry
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    • v.61 no.4
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    • pp.405-410
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    • 2018
  • Pseudomonas tolaasii strain No. 6264 has been isolated from mushroom tissue and identified as one of the major pathogen causing brown blotch disease. It secretes peptide toxins, known as tolaasin and its analogue peptides. P. tolaasii 6264 has been used as a typical pathogenic strain to study the brown blotch disease for last 20 years after confirming its blotch-forming ability, hemolytic activity, and white line formation. In this study, the characteristics of P. tolaasii 6264 strain were analyzed and compared according to storage period. Strains of P. tolaasii 6264 stored annually since 2012 were cultured and their pathogenic characters were analyzed. When the 16S rRNA sequences were compared, all strains were divided into two groups. Pathogenic characters including hemolytic activity, blotch-forming ability, and white line test were also investigated. The strains, P. tolaasii 6264-15-2 and P. tolaasii 6264-17, had all three activities; however, the rest of stored strains showed only blotch-forming ability losing other pathogenic characters. Tolaasin peptides were purified from the bacterial cultures and analyzed by mass spectrometry. The strains, P. tolaasii 6264-15-2 and P. tolaasii 6264-17, secreted Tol I (1987 Da), Tol II (1943 Da), and its analogues (1973 Da, 2005 Da) while some of these peptides were not found in the media cultured other strains. These results indicate that the pathogenicity of P. tolaasii could be varied during the storage period.