• Title/Summary/Keyword: restriction analysis

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Genomic Species Identification of Acinetobacter calcoaceticus - Acinetobacter baumannii Complex Strains by Amplified Ribosomal DNA Restriction Analysis (ARDRA) (Amplified Ribosomal DNA Restriction Analysis (ARDRA) 방법을 이용한 국내 분리 Acinetobacter calcoaceticus - Acinetobacter baumannii Complex 균주의 유전자종 동정)

  • Oh, Jae-Young;Cho, Jae-We;Park, Jong-Chun;Lee, Je-Chul
    • The Journal of the Korean Society for Microbiology
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    • v.35 no.1
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    • pp.69-76
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    • 2000
  • Members of the genus Acinetobacter are recognized as newer pathogens of the nosocomial infection with an increasing frequency in recent years. Strains that belonged to A. calcoaceticus A. baumannii complex (genomic species 1, 2, 3, and 13TU) were major groups associated with nosocomial infection. Phenotypic identification was unreliable and laborious method to classify Acinetobacter strains into 19 genomic species. Rapid and reliable identification of clinical isolates is essential to diagnosis and epidemiology of Acinetobacter. We investigated the suitability of amplified ribosomal DNA restriction analysis (ARDRA) to identify genomic species of 131 Acinetobacter isolates. The 16S rRNA genes (ribosomal DNA) were enzymatically amplified and the amplified PCR products were restricted independently with the enzymes, AluI, CfoI, and MboI. Genomic species of Acinetobacter was classified by the combinations of restriction patterns. The analysis was showed that restriction profiles were characteristic for each genomic species. One hundred fourteen isolates were identified as A. baumannii, twelve were identified as genomic species 13TU, and one was identified as genomic species 3. Four isolates were found to be unknown organisms. All of the isolates which were identified to A. baumannii by phenotypic tests were completely discriminated into A. baumannii and genomic species 13TU by ARDRA. This study demonstrates that ARDRA is a rapid and simple techniques for the identification of Acinetobacter species according to the genomic species.

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Phylogeny of Ganoderma Based on the Restriction Enzyme Analysis of Mitochondrial DNA (미토콘드리아 DNA의 제한효소 분석법에 의한 영지의 계통분류)

  • Hong, Soon-Gyu;Jung, Hack-Sung
    • Korean Journal of Microbiology
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    • v.32 no.4
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    • pp.245-251
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    • 1994
  • Ten strains of 7 species from the genus Ganoderma, G. lucidum ATCC 64251, FP-103561-T, and ES70701, G. applanatum ATCC 44053 and FP-57035-T. G. lobatum ATCC 42985, G. resinaceum ATCC 52416, G. subamboinense var. laevisporum ATCC 52420, G. meredithae ATCC 64492, and G. microsporum ATCC 76024, were studied to discuss their phylogenetic relationships by utilizing restriction fragment length polymorphisms (RFLPs) of mitochondrial DNAs (mtDNAs). Six restriction enzymes, BamHI, BglII, EcoRI, HindIII, PvuII, and XbaI which digested mtDNAs into adequate numbers of restriction fragments for cluster analysis, were used in this study. Restriction profiles of strains for each restriction enzyme were treated as analysis characters to calculate similarity coefficients, which were converted into nucleotide sequence divergence values whose mean values were then arranged in a matrix table. This table was utilized for a phylogenetic analysis using the Neighborjoining method of the PHYLIP package to construct phylogenetic tree. Three strains of G. lucidum and two strains of G. applanatum exhibited different lineages each but one of G. applanatum strains showed a close relationship with G. lobatum, which reflected the species complexity of these species whose strains were phenotypically indistinguishable but genetically distinct. The present results suggest that the natural classification of Ganoderma needs to be considered from the viewpoints of molecular biology-based systematics as well as morphological classifications and cultural identifications for better phylogenetic conclusions.

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The Effects of Activity and Family Support on the Participation Restriction of Chronic Stroke Patients (만성 뇌졸중 환자의 참여제한에 활동과 가족지지가 미치는 영향)

  • Kim, Won-Ho
    • Physical Therapy Korea
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    • v.19 no.1
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    • pp.76-85
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    • 2012
  • The purpose of this study was to identify the factors determining the participation restriction of chronic stroke patients based on international classification of functioning, disability, and health (ICF) model. Sixty-eight stroke patients participated. The participants were assessed participation restriction using the Korean version of London handicap scale (K-LHS), modified Barthel index (K-MBI) to measure activities of daily living, Berg balance scale (K-BBS) to assess balance, and the center for epidemiologic studies depression (K-CES-D) to gauge depression. Also, 3 minutes walking test (3MWT), gait velocity, asymmetric posture, and family support were assessed. A stepwise multiple regression analysis was used to explore the factors determining participation restriction. There were no significant different in the K-LHS and K-MBI results by gender (p>.05). Correlations between the K-LHS and K-MBI (r=-.656), K-BBS (r=-.543), K-CES-D (r=.266), 3MWT (r=-.363), gait velocity (r=.348), and family support (r=-.389) were significant (p<.05). Also, the K-MBI and family support were the factors that determined participation restriction (p<.05) and that 40.2% of the variation in the K-LHS can be explained. Therefore, it is suggested that evaluation and intervention of patient's activity level and extent of family support is necessary to reduce participation restriction of chronic stroke patients.

POLYMERASE CHAIN REACTION AND RESTRICTION FRAGMENT LENGTH POLYMORPHISM OF 16S RIBOSOMAL DNA OF STREPTOCOCCI ISOLATED FROM INFECTED ROOT CANALS (감염 근관에서 분리된 연쇄구균의 16S Ribosomal DNA 중합효소 연쇄반응과 제한효소 절단길이 다형성에 관한 연구)

  • Jung, Hee-Il;Im, Mi-Kyung
    • Restorative Dentistry and Endodontics
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    • v.20 no.2
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    • pp.577-609
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    • 1995
  • Bacteria have been regarded as one of the most important factors in pulpal and periapical diseases. Streptococci are frequently isolated facultative anaerobes in infected root canals. Recently molecular biological techniques have been rapidly progressed. This study was designed to apply the molecular biological tools to the identification and classification of streptococci in the endodontic microbiology. Streptococci isolated from infected root canals were identified with both Vitek Systems and API 20 STREP. Identification results were somewhat different in several strains of streptococci. Eighteen streptococci and enterococcal was difficult so to digest plasmid DNA using Hind III and EcoRI to differentiate strains by restriction enzyme analysis of plasmid DNA. 16S rDNA of chromosome was amplified by polymerase chain reaction(PCR) and then restricition fragment length polymorphism(RFLP) using several restriction enzymes was observed. The molecular mass of 16S rDNA of chromosomal DNA was approximately 1.4kb. There were three to five RFLP patterns using eight restriction enzymes. RFLP patterns digested with CfoI which recognizes four base sequences were identical in all stains. Hind III which recognizes six base sequences could not digest the 16S rDNA. Restriction enzymes which recognize five base sequences were suitable for RFLP pattern analysis. At least three different restriction enzymes were needed to compare each strains. 16S rDNA PCR-RFLP was simple and rapid to differentiate and classify strains and could be used in the epidemiological study of root canal infections.

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Use of Restriction Fragment Length Polymorphism Analysis to Differentiate Fungal Strains in Sunchang Meju

  • Jung, Jong-Hyun;Seo, Dong-Ho;Bhoo, Sung-Hee;Ha, Suk-Jin;Kim, Jong-Sang;Kim, Jeong-Hwan;Kwon, Dae-Young;Cha, Jae-Ho;Park, Cheon-Seok
    • Food Science and Biotechnology
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    • v.17 no.4
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    • pp.888-891
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    • 2008
  • Twenty-three fungal strains were isolated from meju that had originated from the Sunchang province, the famous location for making fermented soybean foods in Korea. The restriction fragment length polymorphism (RFLP) of the internal transcribed spacer (ITS) region of the rDNA (ITS-RFLP) was applied to differentiate the isolated fungal strains. First, the ITS region by polymerase chain reaction (PCR) with specific primers was amplified and then cleaved the products with different restriction enzymes. Cleavage of the amplified fragments with the restriction enzymes AluI, HaeIII, HhaI, and TaqI revealed extensive polymorphisms. The ITS-RFLP results highly correlated with ITS sequence analysis. All of the 23 fungal strains were classified into 5 groups by ITS-RFLP analysis. Aspergillus oryzae was the major fungal strain isolated from Sunchang meju (12 out of 23), while Aspergillus fumigatus was the next most frequently isolated strain (7 out of 23). In contrast, it was found that Fusarium asiaticum, Aspergillus sydowii, and Arthrinium sp. were the minor fungal strains in meju.

Comparison of Terminal-restriction Fragment Length Polymorphism (T-RFLP) Analysis and Sequencing of 16S rDNA Clones in marine sediments

  • Lee Jung-Hyun
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2002.10a
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    • pp.15-21
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    • 2002
  • Terminal-restriction fragment length polymorphism (T-RFLP) analysis has been optimized by using in vitro model community composed of genomic DNAs of known bacterial strains and has been applied to assess the bacterial community structure in marine sediments. The specific fluorescence-labeled terminal restriction fragments (T-RFs) between 39 and 839 base long specifying each strain were precisely measured for known bacterial strains. The addition of a co-solvent (dimethylsulfoxide or glycerol) into PCR reactions has reduced differential PCR amplification. Comparative bacterial community structure was investigated for pristine and polluted sediments. A complex T-RFLP pattern showing complex bacterial community structure was obtained in the pristine sediment, whereas simple T-RFLP pattern (low bacterial diversity) was shown in polluted sediments where caged aquaculture has been conducted for several years. The results of T-RFLP analysis were compared with that of cloning and sequencing 16S rDNA clones from the same sediments. Sequence analysis of 16S rDNA clones (72) of the pristine sediment revealed a diverse collection of lineages, largely of the class Proteobacteria ($6\%$ alpha subdivision, $46\%$ gamma subdivision, $13\%$ delta subdivision, and $3\%$ epsilon subdivision), Nitrospina $(8\%)$, high G+C gram positive $(8\%)$, Verrucomicrobia $(7\%)$, and Planctomycetes $(6\%)$. In the contaminated sediments, 17 $(59\%)$ of the 16S rDNA clones (29) were related to Campylobacter and symbiont of Rimicaris exoculata belonging to epsilon subdivision of Proteobacteria. The results obtained indicated that T-RFLP analysis is a rapid and precise technique for comparative bacterial community analysis.

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A Study on Chargin and Discharging Characteristics of Variable Volume with Compressed Air (가변체적내의 압축공기 충진 및 방출특성연구)

  • Kim, Dong-Soo;Kim, Hyoung-Eui;Park, Jae-Bum;Kang, Bo-Sig;Sung, Baek-Ju
    • Journal of the Korean Society for Precision Engineering
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    • v.12 no.11
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    • pp.125-131
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    • 1995
  • Pneumatic System has been mainly used as main equipment for actuation and control of fluid force in manufacturing industry. For velocity control of piston, meter-out restriction method is used in many cases. In this systems, meter-out restriction method is adopted for analysing the Dynamic Charging and Discharging Process which is Variable Volume Chamber. Experiments has been conducted for different supply pressure condition. As a experimental result, charge side chamber pressure rises to supply pressure rapidily and discharge side chamber pressure decreases. Also, when the air in the cylinder is discharged, tempdrature of air decreases steeply. Restriction of the Cylinder sometimes freeze and it dose not function. The result will be useful for the analysis of pneumatic system.

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The Characterization of Mitochondrial DNA of Korean Ginseng (Panax ginseng C.A. Meyer)

  • Lim, Yong-Pyo;Park, Kwang-Tae
    • Proceedings of the Ginseng society Conference
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    • 1990.06a
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    • pp.168-174
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    • 1990
  • This study was focused on the characterization of mitochondrial DNA (mtDNA) for molecular genetically approach of energy Production related mechanism in Panax Ein.fend. The simple and efficient method of mtDNA isolation from ginseng has been developed by modification of recently advanced methods. This procedure can successfully apply to mtDNA isolation of several plants. MtDNA of etiolated shoot and one-year root were digested with restriction endonucleases, but that of 6-year root not Any difference was not observed in the restriction endonuclease digestion patterns among the ginseng variants. Molecular size of ginseng mtDNA was estimated at least 159 kb by the restriction endonuclease fragment analysis. The 4.5 kb extra band at the lane of EcoRll treatment could be observed in restriction patterns digested with the methylation sensitive endonucleases, BstN 1 and EcoRll. For construction of mitochondrial genomic library of ginseng, mtDNA was partially digested with EcoRl, and packaged with EMBL4 phage vector Genomic library was screened and purified for further research including restricttion mapping of ginseng mtDNA, and cloning of the genes. The gene of ATP synthase A subunit was cloned koto the purified EMBL4 library clone No. 16. Now, clone No. 16 is subcloned for structure gene sequence analysis.

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Monitoring of Microorganisms Added into Oil-Contaminated Microenvironments by Terminal-Restriction Fragment Length Polymorphism Analysis

  • JUNG SEONG-YOUNG;LEE JUNG-HYUN;CHAI YOUNG-GYU;KIM SANG-JIN
    • Journal of Microbiology and Biotechnology
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    • v.15 no.6
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    • pp.1170-1177
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    • 2005
  • Terminal-restriction fragment length polymorphism (T-RFLP) analysis was used to monitor inoculated oil-degrading microorganisms during bioremedial treatability tests. A pair of universal primers, fluorescently labeled 521F and 1392R, was employed to amplify small subunit rDNA in order to simultaneously detect two bacterial strains, Corynebacterium sp. IC10 and Sphingomonas sp. KH3-2, and a yeast strain, Yarrowia lipolytica 180. Digestion of the 5'-end fluorescence/labeled PCR products with HhaI produced specific terminal-restriction fragments (T-RFs) of 185 and 442 bases, corresponding to Corynebacterium sp. IC10 and Y. lipolytica 180, respectively. The enzyme NruI produced a specific T-RF of 338 bases for Sphingomonas sp. KH3-2. The detection limit for oildegrading microorganisms that were inoculated into natural environments was determined to be $0.01\%$ of the total microbial count, regardless of the background environment. When three oil-degrading microorganisms were released into oil-contaminated sand microenvironments, strains IC10 and 180 survived for 35 days after inoculation, whereas strain KH3-2 was detected at 8 days, but not at 35 days. This result implies that T-RFLP could be a useful tool for monitoring the survival and relative abundance of specific microbial strains inoculated into contaminated environments.

A Study on Characteristics of Urban Landscape using Digital Elevation Model and Land Use Data -The Case of Chunchon City- (수치표고모델과 토지이용 데이터를 이용한 광역 도시경관의 특성에 관한 연구 -강원도 춘천시를 사례로-)

  • 한갑수
    • Journal of Environmental Science International
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    • v.12 no.10
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    • pp.1025-1032
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    • 2003
  • The purpose of this study is to analyze the characteristics of the urban landscape by CG(Computer Graphic) pictures and visibility analysis. In this study, the CG picture was created by using zoning data and DEM(Digital Elevation Model) on Chunchon city. The landscape type was classified into three by cluster analysis using the area rate of the element which constitutes CG picture. Visible analysis used DEM as fundamental data, and was performed by using GIS(Geographic Information System). And the frequency seen on viewshed was classified into five grades. The result of this study are as follows: 1. The area rate of the element which constitutes CG picture was high development restriction zone(43%) in type 1, green land area(39%) in type 2, and city area(24%) in type 3. 2. In the analysis results of GIS visibility analysis, it turns out that Euiam lake is important area on the landscape in all types. 3. The land use of high frequency seen on viewshed was water area, residential area in type 1, development restriction area, park area in type 2, and development restriction zone, water area in type 3.