• Title/Summary/Keyword: 16s rDNA

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Molecular Taxonomy based on 16S rDNA Analysis and Pathogenicity of Yersinia pseudotuberculosis Isolated from Spring Waters (약수에서 분리한 Yersinia pseudotuberculosis의 병원성과 16S rDNA 분석에 의한 분자학적 분류)

  • Lee, Young-Kee;Choi, Sung-Min;Oh, Soo-Kyung;Lee, Kang-Moon;Ryeom, Kon
    • Korean Journal of Microbiology
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    • v.37 no.1
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    • pp.9-14
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    • 2001
  • In order to investigate the pathogenicity and development of differential identification technique in the Yersinia species and other entericbacteria, we isolated 5 strains of Y.pseudotuberculosis from spring water sites in Seoul. The biochemical characteristics of isolated strains revealed that indole, VP($25^{\circ}C$, $37^{\circ}C$), $H_2S$, phenylalanine, lysine, arginine, ornithine, gas from glucose, lactose, sucrose, sorbitol, oxidase and motility($37^{\circ}C$) were all negative and urease, glucose, mannitol, salicin, catalase and motility($25^{\circ}C$) were all positive. To detect the causative agent of pseudotuberculosis(Y.pseudotuberculosis), we carried out a study using a PCR with inv primers complementary to the pathogenic region and found that all strains were positive, this revealed that strains from spring waters were pathogenic. Also 16S rDNA for total 5 strains of Y. pseudotuberculosis were amplified and a stretch of approximately 1,450 nucleotides were sequenced and analyzed. The 16S rDNA nucleotide sequence homologies among Yersinia species ranged 97.5% to 100% and between Y.pseudotuberculosis and other entericbacteria they ranged 93.0% to 95.1%. The Phylogenetic tree generated from the sequence analysis of the 16S rDNA gene showed 3 coherent clusters that could be separated into Y.pseudotuberculsis strains, some Yersinia species strains and other entericbacteria strains.

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Molecular Differentiation of Bacillus spp. Antagonistic Against Phytopathogenic Fungi Causing Damping-off Disease

  • Cho, Min-Jeong;Kim, Young-Kwon;Ka, Jong-Ok
    • Journal of Microbiology and Biotechnology
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    • v.14 no.3
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    • pp.599-606
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    • 2004
  • Gram-positive antagonistic bacilli were isolated from agricultural soils for possible use in biocontrol of plant pathogenic fungi, Fusarium oxysporum, Rhizoctonia solani, and/or Pythium ultimum. Among the 65 antagonistic Gram-positive soil isolates, 22 strains were identified as Bacillus species by 16S rDNA sequence analyses. Four strains, including DF14, especially exhibited multiple antagonistic properties against the three damping-off fungi. Genotypic properties of the Bacillus isolates were characterized by rapid molecular fingerprinting methods using repetitive extragenic palindromic-PCR (REP-PCR), ribosomal intergenic spacer-length polymorphisms (RIS-LP), 16S rDNA PCR-restriction fragment length polymorphisms (PCR-RFLP), and strain-specific PCR assays. The results indicated that the REP-PCR method was more valuable than the RIS-LP and 16S rDNA PCR-RFLP analyses as a rapid and reliable approach for bacilli typing and identification. The use of strain-specific primers designed based on 16S rDNA sequence comparisons enabled it to be possible to selectively detect a strain, DF14, which is being used as a biocontrol agent against damping-off fungi.

Molecular Discrimination of Mitis Group Streptococci Isolated from Koreans using RpoB Nucleotide Sequences

  • Park, Soon-Nang;Kook, Joong-Ki
    • International Journal of Oral Biology
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    • v.38 no.1
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    • pp.29-36
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    • 2013
  • Mitis group streptococci (MGS) were classified based on the nucleotide sequences 16S rRNA gene (16S rDNA) and comprised 13 Streptococcus species. However, 16S rDNA homogeneity among MGS was too high to discriminate between clinical strains at the species level, notably between Streptococcus mitis, Streptococcus oralis, Streptococcus pneumoniae, and Streptococcus pseudopneumoniae. The purpose of this study was to discriminate between 37 strains of MGS isolated from Korean oral cavities using phylogenetic analysis of the DNA-dependant RNA polymerase beta-subunit gene (rpoB). 16S rDNA and rpoB from clinical strains of MGS were sequenced using the dideoxy chain termination method and analyzed using MEGA version 5 software. The resulting phylogenetic data showed that the rpoB sequences could delineate clinical strains of MGS at the species level. Phylogenetic analysis of rpoB is therefore a useful approach for identifying MGS at the species level.

Diversity of Pigment-Producing Halophilic Bacteria Isolated from Coastal Seawater and Solar Saltern in Korea (한국 근해와 염전에서 분리한 색소 생성 호염성 세균의 다양성)

  • Yong Hae-Young;Park Jin-Sook
    • Korean Journal of Microbiology
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    • v.40 no.4
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    • pp.302-306
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    • 2004
  • A total of forty strains of pigment-producing halophilic bacteria were isolated from the solar saltern and coastal seawater in Korea. The diversity of those bacteria were determined on the basis of PCR-RFLP and 16S rDNA sequences. The isolated strains were clssified into nine genera: Pseudoalteromonas, Photobacterium, Vibrio, Halobacillus, Bacillus, Paracoccus, Salinicoccus, Tenacilbaculum, and Flavobacterium. While more than $80\%$ of the pigment-producing halophilic bacteria isolated from the coastal seawater were classified as gram-negative Pseudolateromonas, most of the strains isolated from the solar saltern were classified into gram-positive Halo­bacillus. The other strain was KK7, which may be identified as novel species belonging to the genus, Salini­coccus.

The List of Korean Organisms Registered in the NCBI Nucleotide Database for Environmental DNA Research (환경유전자 연구를 위한 NCBI Nucleotide 데이터베이스에 등록된 국내 생물 목록 현황)

  • Ihn-Sil Kwak;Chang Woo Ji;Won-Seok Kim;Dongsoo Kong
    • Korean Journal of Ecology and Environment
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    • v.55 no.4
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    • pp.352-359
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    • 2022
  • Recently, with the development of genetic technology, interest in environmental DNA (eDNA) to study biodiversity according to molecular biological approaches is increasing. Environmental DNA has many advantages over traditional research methods for biological communities distributed in the environment but highly depends on the established base sequence database. This study conducted a comprehensive analysis of the habitat status and classification at the genus level, which is mainly used in eDNA (12S rRNA, 16S rRNA, 18S rRNA, COI, and CYTB), focusing on Korean registration taxon groups (phytoplankton, zooplankton, macroinvertebrates, and fish). As a result, phytoplankton and zooplankton showed the highest taxa proportion in 18S rRNA, and macroinvertebrates observed the highest ratio in the nucleotide sequence database in COI. In fish, all genes except 18S rRNA showed a high taxon ratio. Based on the Korean registration taxon group, the gene construction of the top 20 genera according to bio density observed that most of the phytoplankton were registered in 18S rRNA, and the most significant number of COI nucleotide sequences were established in macroinvertebrates. In addition, it was confirmed that there is a nucleotide sequence for the top 20 genera in 12S rRNA, 16S rRNA, and CYTB in fish. These results provided comprehensive information on the genes suitable for eDNA research for each taxon group.

Specific Primer for Detection of Jujube Witches' Broom Phytoplasma Group (16SrV) in Korea

  • Han, Sang-Sub
    • The Plant Pathology Journal
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    • v.21 no.1
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    • pp.55-58
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    • 2005
  • In order to diagnose and differentiate jujube witches' broom (JWB) phytoplasma rapidly, oligonucleotide primer pair, 16Sr(V) F/R, for polymerase chain reactions (PCRs) was designed on the basis of 16S rRNA sequences of JWB phytoplasma. The PCR employing phytoplasma universal primer pair P1/P7 consistently amplified DNA in all tested phytoplasma isolates. But no phytoplasma DNA was detected from healthy jujube seedlings. The nested PCR, the primer pair 16S(V) F/R, about 460 bp fragment, amplified DNA in all tested JWB and related phytoplasmas including ligustrum witches' broom phytoplasma of the 16S rRNA group V, but no DNA amplification was detected from other phytoplasma strains such as groups 16SrI (Aster yellows) and 16SrXII (Stolbur group) in which mulberry dwarf phytoplasma and chrysanthemum witches' broom phytoplasma belong to, respectively. The same results were obtained from both Korean and Chinese isolates of JWB phytoplasma. Nested-PCR using phytoplasma universal primer pair P1/P7 and 16SrV group-specific primer pair 16S(V) F/R could detect group V phytoplasmas rapidly and easily, in particular JWB phytoplasma.

Comparison between DNA- and cDNA-based gut microbial community analyses using 16S rRNA gene sequences (16S rRNA 유전자 서열 분석을 이용한 DNA 및 cDNA 기반 장내 미생물 군집 분석의 비교)

  • Jo, Hyejun;Hong, Jiwan;Unno, Tatsuya
    • Korean Journal of Microbiology
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    • v.55 no.3
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    • pp.220-225
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    • 2019
  • Studies based on microbial community analyses have increased in the recent decade since the development of next generation sequencing technology. Associations of gut microbiota with host's health are one of the major outcomes of microbial ecology filed. The major approach for microbial community analysis includes the sequencing of variable regions of 16S rRNA genes, which does not provide the information of bacterial activities. Here, we conducted RNA-based microbial community analysis and compared results obtained from DNA- and its cDNA-based microbial community analyses. Our results indicated that these two approaches differed in the ratio of Firmicutes and Bacteroidetes, known as an obesity indicator, as well as abundance of some key bacteria in gut metabolisms such as butyrate producers and probiotics strains. Therefore, cDNA-based microbial community may provide different insights regarding roles of gut microbiota compared to the previous studies where DNA-based microbial community analyses were performed.

Diversity of Halophilic Archaea From Six Hypersaline Environments in Turkey

  • Ozcan, Birgul;Ozcengiz, Gulay;Coleri, Arzu;Cokmus, Cumhur
    • Journal of Microbiology and Biotechnology
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    • v.17 no.6
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    • pp.985-992
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    • 2007
  • The diversity of archaeal strains from six hypersaline environments in Turkey was analyzed by comparing their phenotypic characteristics and 16S rDNA sequences. Thirty-three isolates were characterized in terms of their phenotypic properties including morphological and biochemical characteristics, susceptibility to different antibiotics, and total lipid and plasmid contents, and finally compared by 16S rDNA gene sequences. The results showed that all isolates belong to the family Halobacteriaceae. Phylogenetic analyses using approximately 1,388 bp comparisions of 16S rDNA sequences demonstrated that all isolates clustered closely to species belonging to 9 genera, namely Halorubrum (8 isolates), Natrinema (5 isolates), Haloarcula (4 isolates), Natronococcus (4 isolates), Natrialba (4 isolates), Haloferax (3 isolates), Haloterrigena (3 isolates), Halalkalicoccus (1 isolate), and Halomicrobium (1 isolate). The results revealed a high diversity among the isolated halophilic strains and indicated that some of these strains constitute new taxa of extremely halophilic archaea.

Application of 16S rDNA PCR-RFLP Analysis for the Rapid Identification of Weissella Species (Weissella 속 유산균의 빠른 동정을 위한 16S rDNA PCR-RFLP 분석법의 적용)

  • Lee, Myeong-Jae;Cho, Kyeung-Hee;Lee, Jong-Hoon
    • Microbiology and Biotechnology Letters
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    • v.38 no.4
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    • pp.455-460
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    • 2010
  • A polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) analysis was applied to detect and identify ten Weissella spp. frequently found in kimchi. The previously reported genus-specific primers designed from 16S rDNA sequences of Weissella spp. were adopted but PCR was performed at the increased annealing temperature by $4^{\circ}C$. The sizes of amplified PCR products and restricted fragments produced by AluI, MseI, and BceAI endonucleases were well correspond with the expected sizes. W. kandleri, W. koreensis, W. confusa, W. minor, W. viridescens, W. cibaria, and W. soli were distinguished by AluI and MseI and W. hellenica and W. paramesenteroides were identified by BceAI. W. thailandensis was distinguished when restriction pattern of other species was compared but identified by the single use of MspI.

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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