• Title/Summary/Keyword: 16S rDNA sequence analysis

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Microbial Genome Analysis and Application to Clinical Bateriology (미생물의 유전자(Genome) 해석과 임상세균학에 이용)

  • Kim, Sung-Kwang
    • Journal of Yeungnam Medical Science
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    • v.19 no.1
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    • pp.1-10
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    • 2002
  • With the establishment of rapid sequence analysis of 16S rRNA and the recognition of its potential to determine the phylogenetic position of any prokaryotic organism, the role of 16S rRNA similarities in the present species definition in bacteriology need to be clarified. Comparative studies clearly reveal the limitations of the sequence analysis of this conserved gene and gene product in the determination of relationship at the pathogenic strain level for which DNA-DNA reassociation experiments still constitute the superior method. Since today the primary structure of 16S rRNA is easier to determine than hybridization between DNA strands, the strength of the sequence analysis is to recognize the level at which DNA pairing studies need to be performed, which certainly applies to similarities of 97% and higher.

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Identification of Bacteria Causing Fermentation of Oriental Melon in Korea (참외 발효과를 유발하는 세균의 동정)

  • Choi, Jae-Eul;Cha, Sun-Kyung;Kim, Jin-Hee;Yuk, Jin-Ah;Hwang, Yong-Soo;Kwon, Soon-Wo
    • Research in Plant Disease
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    • v.9 no.4
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    • pp.189-195
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    • 2003
  • Bacteria causing fermentation in Oriental melon were identified as three independent groups on the basis of 16S rDNA sequence analysis. The 16S rDNA sequence of the strain CM2105 showed the highest identity (99.6%) with that of Microbacterium phyllosphaerae, and also indicated high sequence identity to that of M. holiorum (99.5%). The 16S rDNA sequences of the strain CM2101 and CM2121 matched at the high sequence similarity (98.9%, 98.8, respectively), to that of Pseudomonas pavonacea, and the DNA sequence of CM2126 showed high sequence identity to that of P. costantinii (99.5%), and P. grimontii (99.0%). The 16S rDNA sequence of the strain CM2113 showed the highest identity (99.7%) with that of Enterobacter cloacae. The 16S rDNA sequences, the physiological and biochemical analysis suggested that the strain CM2105 belonged to Microbacterium phyllosphaerae, CM2101, CM2121 and CM2126 to Pseudomonas spp., CM2113 to Enterobacter cloacae.

Diversity of Myxobacteria in Soil Samples from Asansi and Uponeup in Korea (아산시와 우포늪 토양의 점액세균 다양성)

  • Chung, Jin-Woo;Kim, Jin-Woo;Cho, Kyung-Yun
    • Korean Journal of Microbiology
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    • v.46 no.4
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    • pp.405-408
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    • 2010
  • Diversity of myxobacteria in five soil samples from Asansi and Uponeup in Korea was explored by means of polymerase chain reaction (PCR) using primers that specifically bind 16S rDNA of myxobacteria. DNA sequence analysis of 76 PCR fragments containing myxobacterial 16S rDNA revealed five putative novel myxobacterial genera whose 16S rDNA sequences shared <95% sequence identity with those of the type strains. This finding indicates the presence of many uncultured and unidentified myxobacterial species in Korean soil.

Physiological and Phylogenetic Analysis of Burkholderia sp. HY1 Capable of Aniline Degradation

  • Kahng, Hyung-Yeel;Jerome J. Kukor;Oh, Kye-Heon
    • Journal of Microbiology and Biotechnology
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    • v.10 no.5
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    • pp.643-650
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    • 2000
  • A new aniline-utilizing microorganism, strain HY1 obtained from an orchard soil, was characterized by using the BIOLOG system, an analysis of the total cellular fatty acids, and a 16S rDNA sequence. Strain HY1 was identified as a Burkholderia species, and was designated Burkholderia sp. HY1. GC and HPLC analyses revealed that Burkholderia sp. HY1 was able to degrade aniline to produce catechol, which was subsequently converted to cis,cis-muconic acid through an ortho-ring fission pathway under aerobic conditions. Strain HY1 exhibited a drastic reduction in the rate of aniline degradation when glucose was added to the aniline media. However, the addition of peptone or nitrate to the aniline media dramatically accelerated the rate of aniline degradation. A fatty acid analysis showed that strain HY1 was able to produce lipids 16:0 2OH, and 11 methyl 18:1 ${\omega}7c$ approximately 3.7-, 2.2-, and 6-fold more, respectively, when grown on aniline media than when grown on TSA. An analysison the alignment of a 1,435 bp fragment. A phylogenetic analysis of the 16S rDNA sequence based on a 1,420 bp multi-alignment sowed of the 16s rDNA sequence revealed that strain HY1 was very closely related to Burkholderia graminis with 95% similarity based that strain HY1 was placed among three major clonal types of $\beta$-Proteobacteria, including Burkholderia graminis, Burkholderia phenazinium, and Burkholderia glathei. The sequence GAT(C or G)${\b{G}}$, which is highly conserved in several locations in the 16S rDNA gene among the major clonal type strains of $\beta$-Proteobacteria, was frequently replaced with GAT(C or G)${\b{A}}$ in the 16S rDNA sequence from strain HY1.

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Isolation of Lactococci Inhibiting Listeria monocytogenes from Kimchi Habitat and Its Identification by 16S rDNA Analysis (김치 서식처에서 Listeria monocytogenes를 억제하는 lactococci의 분리와 16S rDNA분석에 의한 동정)

  • 박은주;한홍의;민봉희
    • The Korean Journal of Ecology
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    • v.22 no.1
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    • pp.45-50
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    • 1999
  • A bacteriocin-producing strain was isolated from kimchi at the early stage of kimchi fermentation. It was identified as Lactococcus lactis by morphological, cultural and physiological characteristics and partial sequence of 16S rDNA. The bacteriocin from isolate had antimicrobial activity against gram positive pathogenic bacteria, such as Listeria monocytogenes. Staphylococcus aureus and several strains of lactic acid bacteria but not to gram negative bacteria, Yersinia enterocolitica. The bacteriocin was sensitive to protease, protease ⅩⅣ, a-chymotrypsin and pepsin but not to lipase, trypsin and lysozyme. The bacteriocin activity was stable at pH 2-11 and temperature of 100 for 10 min. Thus, Listeria monocytogenes could be inhibited by Lactococcus lactis at early stage of fermentation.

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Keratitis by Acanthamoeba triangularis: Report of Cases and Characterization of Isolates

  • Xuan, Ying-Hua;Chung, Byung-Suk;Hong, Yeon-Chul;Kong, Hyun-Hee;Hahn, Tae-Won;Chung, Dong-Il
    • Parasites, Hosts and Diseases
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    • v.46 no.3
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    • pp.157-164
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    • 2008
  • Three Acanthamoeba isolates (KA/E9, KA/E17, and KA/E23) from patients with keratitis were identified as Acanthamoeba triangularis by analysis of their molecular characteristics, a species not previously recognized to be a corneal pathogen. Epidemiologic significance of A. triangularis as a keratopathogen in Korea has been discussed. Morphologic features of Acanthamoeba cysts were examined under a microscope with differential interference contrast (DIC) optics. Mitochondrial DNA (mtDNA) of the ocular isolates KA/E9, KA/E17, and KA/E23 were digested with restriction enzymes, and the restriction patterns were compared with those of reference strains. Complete nuclear 188 and mitochondrial (mt) 16S rDNA sequences were subjected to phylogenetic analysis and species identification. mtDNA RFLP of 3 isolates showed very similar patterns to those of SH621, the type strain of A. triangularis. 16S and 18S rDNA sequence analysis confirmed 3 isolates to be A. triangularis. 18S rDNA sequence differences of the isolates were 1.3% to 1.6% and those of 16S rDNA, 0.4% to 0.9% from A. triangularis SH621. To the best of our knowledge, this is the first report, confirmed by 18S and 16S rDNA sequence analysis, of keratitis caused by A. triangularis of which the type strain was isolated from human feces. Six isolates of A. triangularis had been reported from contaminated contact lens cases in southeastern Korea.

Application of rDNA-PCR Amplification and DGGE Fingerprinting for Detection of Microbial Diversity in a Malaysian Crude Oil

  • Liew, Pauline Woan Ying;Jong, Bor Chyan
    • Journal of Microbiology and Biotechnology
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    • v.18 no.5
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    • pp.815-820
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    • 2008
  • Two culture-independent methods, namely ribosomal DNA libraries and denaturing gradient gel electrophoresis (DGGE), were adopted to examine the microbial community of a Malaysian light crude oil. In this study, both 16S and 18S rDNAs were PCR-amplified from bulk DNA of crude oil samples, cloned, and sequenced. Analyses of restriction fragment length polymorphism (RFLP) and phylogenetics clustered the 16S and 18S rDNA sequences into seven and six groups, respectively. The ribosomal DNA sequences obtained showed sequence similarity between 90 to 100% to those available in the GenBank database. The closest relatives documented for the 16S rDNAs include member species of Thermoincola and Rhodopseudomonas, whereas the closest fungal relatives include Acremonium, Ceriporiopsis, Xeromyces, Lecythophora, and Candida. Others were affiliated to uncultured bacteria and uncultured ascomycete. The 16S rDNA library demonstrated predomination by a single uncultured bacterial type by >80% relative abundance. The predomination was confirmed by DGGE analysis.

Isolation and Characterization of Thermophilic Microorganism Producing Starch-hydrolyze Enzyme (한국 토양으로부터 전분가수분해효소를 생산하는 고온성 균주의 선별과 동정)

  • Choi, Wonseok;Bai, Dong-Hoon
    • Food Engineering Progress
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    • v.14 no.1
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    • pp.7-13
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    • 2010
  • A thermophilic microorganism, which is able to hydrolyze starch, was isolated from soil and compost in Korea. It was Gram-positive, rod-shaped, catalase positive, nonmotile, glucose and mannitol fermentative, xylose oxidative, and spore forming microorganism. It also has an ability to hydrolyze casein and gelatin. The color of colony was yellowish white. The sequence of 16S rDNA of strain 2719 showed 99.5% sequence homology with the sequence of 16S rDNA of Bacillus thermoglucosidasius. On the basis of biochemical and physiological properties and phylogenetic analysis, the isolated strain was named as Bacillus thermoglucosidasius 2719.

Genotypic and Phenotypic Diversity of PGPR Fluorescent Pseudomonads Isolated from the Rhizosphere of Sugarcane (Saccharum officinarum L.)

  • Rameshkumar, Neelamegam;Ayyadurai, Niraikulam;Kayalvizhi, Nagarajan;Gunasekaran, Paramsamy
    • Journal of Microbiology and Biotechnology
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    • v.22 no.1
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    • pp.13-24
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    • 2012
  • The genetic diversity of plant growth-promoting rhizobacterial (PGPR) fluorescent pseudomonads associated with the sugarcane (Saccharum officinarum L.) rhizosphere was analyzed. Selected isolates were screened for plant growthpromoting properties including production of indole acetic acid, phosphate solubilization, denitrification ability, and production of antifungal metabolites. Furthermore, 16S rDNA sequence analysis was performed to identify and differentiate these isolates. Based on 16S rDNA sequence similarity, the isolates were designated as Pseudomonas plecoglossicida, P. fluorescens, P. libaniensis, and P. aeruginosa. Differentiation of isolates belonging to the same group was achieved through different genomic DNA fingerprinting techniques, including randomly amplified polymorphic DNA (RAPD), amplified ribosomal DNA restriction analysis (ARDRA), repetitive extragenic palindromic (REP), enterobacterial repetitive intergenic consensus (ERIC), and bacterial repetitive BOX elements (BOX) analyses. The genetic diversity observed among the isolates and rep-PCR-generated fingerprinting patterns revealed that PGPR fluorescent pseudomonads are associated with the rhizosphere of sugarcane and that P. plecoglossicida is a dominant species. The knowledge obtained herein regarding the genetic and functional diversity of fluorescent pseudomonads associated with the sugarcane rhizosphere is useful for understanding their ecological role and potential utilization in sustainable agriculture.

Characterization of Korean Erwinia carotovora Strains from Potato and Chinese Cabbage

  • Seo, Sang-Tae;Koo, Jun-Hak;Hur, Jang-Hyun;Lim, Chun-Keun
    • The Plant Pathology Journal
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    • v.20 no.4
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    • pp.283-288
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    • 2004
  • Four Erwinia carotovora strains isolated from potatoes showing blackleg symptoms and rotted Chinese cabbage were analysed by biochemical tests and sequence analysis of 16S rDNA and 16S-23S rRNA intergenic spacer (IGS) regions, and the data were compared to related E. carotovora strains. Based on the results of the biochemical tests and sequence analysis, 2 of the 4 strains were identified as E. carotovora subsp. carotovora (Ecc), whereas the rest strains were distinct from Ecc. The last two strains, HCC3 and JEJU, were biochemically similar to E, carotovora subsp. atroseptica (Eca). However, the results of sequence analysis and Eca-specific PCR assays showed that the strains were distinct from Eca. On the basis of 16S rDNA sequence analysis, HCC3 and JEJU strains were placed in E. carotovora subsp. odorifera and E. carotovora subsp. wasabiae, respectively. The results of sequence analysis and specific PCR assay for Eca indicated that Asian Eca strains were distinct from European Eca strains, although they were phenotycally homogeneous.