• Title/Summary/Keyword: 16S rRNA Gene

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Use of 16S-23S rRNA Intergenic Spacer Region for Rapid Detection of Vibrio fluvialis (16S-23S rRNA Intergenic Spacer Region을 이용한 Vibrio fluvialis의 검출)

  • 강현실;허문수;이제희
    • Korean Journal of Environmental Biology
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    • v.21 no.1
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    • pp.77-85
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    • 2003
  • We have examined the 16S-23S rRNA intergenic spacer regions (ISR) of Vibrio fluvialis. ISRs were PCR amplified, cloned into a plasmid vector and then sequenced. As results of ISR nucleotide sequence analysis, total of 6 clones were isolated depending on the size. The clones were different in both the number and the composition of the tRNA genes, and were designated ISR-A, ISR-E, ISR-El, ISR-lA, ISR-EKV, ISR-EKAV. ISR-A contains $tRNA^{Ala}$; ISR-lA, $tRNA^{Ile}$-$tRNA^{Ala}$; ISR-EKV, $tRNA^{GIu}$-$tRNA^{Lys}$-$tRNA^{Val}$;ISE-EKAV, $tRNA^{GIu}$-$tRNA^{Lys}$-$tRNA^{Ala}$-$tRNA^{Val}$; ISR -E and E1, $tRNA^{GIu}$ clusters. ISR-EKV was shown to be a minor type out of the six ISR types and showed a very limited homology between ISR-EKV from V, fluvialis and ISRa from other Vibrio species. Therefore ISR-EKV sequence was used to design species-specific primers to detect V, fiuvialis from other Vibrio species by PCR reaction. The specificity of the primers was examined using genomic DNA of other Vibrios as templates for PCR reaction. The result showed that PCR can be a useful method to detect V. fluvialis among Vibrio species in a single PCR reaction.

A report on 10 unrecorded bacterial species isolated from the Korean islands in 2022

  • Seung Yeol Shin;Myung Kyum Kim;Yochan Joung;Yi Hyun Jeon;Ji Hye Jeong;Hyun-Ju Noh;Jaeho Song;Heeyoung Kang
    • Journal of Species Research
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    • v.12 no.spc2
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    • pp.54-59
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    • 2023
  • To obtain unrecorded bacterial species from Korean islands, various samples were collected from the islands in 2022. After plating the samples on marine agar or Reasoner's 2A, and incubating aerobically, approximately 1,200 bacterial strains were isolated and identified using 16S rRNA gene sequences. A total of 10 strains showed ≥98.7% 16S rRNA gene sequence similarity with the bacterial species that were validly published but not reported in Korea. The unrecorded bacterial strains belong to three phyla, five classes, 10 orders, 10 families, and 10 genera, which are assigned to Sphingomonas, Falsirhodobacter and Asticcacaulis of the class Alphaproteobacteria; Colwellia and Halomonas of the class Gammaproteobacteria; Chitinophaga of the class Chitinophagia; Chryseobacterium of the class Flavobacteriia; Microlunatus, Zhihengliuella, and Streptomyces of the class Actinomycetia. The details of the unreported species including Gram reaction, colony and cell morphology, biochemical characteristics, and phylogenetic position are also provided in the description of the strains.

A highly efficient computational discrimination among Streptococcal species of periodontitis patients using 16S rRNA amplicons

  • Al-Dabbagh, Nebras N.;Hashim, Hayder O.;Al-Shuhaib, Mohammed Baqur S.
    • Korean Journal of Microbiology
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    • v.55 no.1
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    • pp.1-8
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    • 2019
  • Due to the major role played by several species of Streptococcus in the etiology of periodontitis, it is important to assess the pattern of Streptococcus pathogenic pathways within the infected subgingival pockets using a bacterial specific 16S rRNA fragment. From the total of 50 patients with periodontitis included in the study, only 23 Streptococcal isolates were considered for further analyses, in which their 16S rRNA fragments were amplified and sequenced. Then, a comprehensive phylogenetic tree was constructed and in silico prediction was performed for the observed Streptococcal species. The phylogenetic analysis of the subgingival Streptococcal species revealed a high discrimination power of the 16S rRNA fragment to accurately identify three groups of Streptococcus on the species level, including S. salivarius (14 isolates), S. anginosus (5 isolates), and S. gordonii (4 isolates). The employment of state-of-art in silico tools indicated that each Streptococcal species group was characterized with particular transcription factors that bound exclusively with a different 16S rRNA-based secondary structure. In conclusion, the observed data of the present study provided in-depth insights into the mechanism of each Streptococcal species in its pathogenesis, which differ in each observed group, according to the differences in the 16S rRNA secondary structure it takes, and the consequent binding with its corresponding transcription factors. This study paves the way for further interventions of the in silico prediction, with the main conventional in vitro microbiota identification to present an interesting insight in terms of the gene expression pattern and the signaling pathway that each pathogenic species follows in the infected subgingival site.

A report of 30 unrecorded bacterial species in Korea, isolated from marine ecosystems in 2021

  • Shin, Seung Yeol;Joung, Yochan;Han, Dukki;Jeong, Ji Hye;Jeon, Yi Hyun;Song, Jaeho
    • Journal of Species Research
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    • v.11 no.3
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    • pp.143-154
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    • 2022
  • To obtain unrecorded bacterial species in Korea, various marine samples were collected from Jeollanam-do Province, Korea in 2021. After plating the samples on marine agar and marine R2A agar, and incubating aerobically and anaerobically, approximately 1200 bacterial strains were isolated and identified using 16S rRNA gene sequences. A total of 30 strains showed ≥98.7% 16S rRNA gene sequence similarity with validly published bacterial species but not reported in Korea, indicating that they are unrecorded bacterial species in Korea. The unrecorded bacterial strains belonged to 4 phyla, 7 classes, 13 orders, 19 families, and 22 genera, which were assigned to Azospirllium, Loktanella, and Pseudovibrio of the class Alphaproteobacteria; Grimontia, Halomonas, Marinobacter, Microbulbifer, Photobacterium, Pseudoalteromonas, Pseudidiomarina, Ferrimonas, Shewanella, Simiduia, Thalassotalea, and Vibrio of the class Gammaproteobacteria; Priestia and Enterococcus of the class Bacilli; Persicobacter of the class Cytophagia; Aureivirga of the class Flavobacteriia; Propionigenium and Psychrilyobacter of the class Fusobacteriia; and Tepidibacter of the class Clostridia. The details of the unreported species including Gram reaction, colony and cell morphology, biochemical characteristics, and phylogenetic position are also provided in the description of the strains.

Characterization of the Microbial Diversity in a Korean Solar Saltern by 16S rRNA Gene Analysis

  • Park, Soo-Je;Kang, Cheol-Hee;Rhee, Sung-Keun
    • Journal of Microbiology and Biotechnology
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    • v.16 no.10
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    • pp.1640-1645
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    • 2006
  • We studied the diversity of the halophilic archaea and bacteria in crystallizer ponds of a Korean solar saltern by analyzing 16S rRNA gene libraries. Although diverse halophilic archaeal lineages were detected, the majority (56%) were affiliated with the uncultured and cultured Halorubrum group. Halophilic archaea that have been frequently observed in solar saltern environments previously, such as Halogeometricum, Halococcus, Haloarcula, and Haloferax, were not detected in our samples. The majority of clones (53%) belonged to the Cytophaga-Flavobacterium-Bacteroides and ${\alpha}-,\;{\gamma}-,\;and\;{\delta}-Proteobacteria$ groups, with 47% of the clones being affiliated with ${\gamma}-Proteobacteria$. We also identified new ${\delta}-Proteobacteria$-related bacteria that have not been observed in hypersaline environments previously. Our data show that the diversity of the halophilic archaea and bacteria in our Korean saltern differs from that of solar salterns found in other geographic locations. We also showed by quantitative real-time PCR analysis that bacteria can form a significant component of the microbial community in solar salterns.

16S rDNA-PCR and RFLP Analysis for rapid identification of Spoilage Bacteria from low Salt Cucumber Brine (저염 발효오이로부터 16S rDNA-PCR과 RFLP분석을 통한 부패균의 신속한 확인)

  • 김재호;장혜영
    • KSBB Journal
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    • v.19 no.1
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    • pp.72-77
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    • 2004
  • The aim of this study was to isolate and identify the spoilage bacteria in the low salt cucumber brine. The PCR amplicons comprising a portion of the 16S rRNA gene of the isolated colonies were directly sequenced and the untrimmed whole sequencing results of the unknown strains were aligned with the type strains using BLAST of NCBI. Then Sequence Aligner and Sequence Match of RDP confirmed the outcome. The identified isolates were eight species and belong to three genuses: Clostridium, Lactobacillus, and Bacillus. The RFLP pattern of the 16S rRNA gene of isolates verified the identified species. From now on the complex spoiling process of law salt fermented cucumber could be analyzed using the isolated species individually or with certain combinations.

Functional Analysis and Selection of Second-site Revertant of Escherichia coli 16S rRNA of C770G (Escherichia coli 16S rRNA 상의 770 위치에 염기치환을 가진 변이체 리보솜의 단백질 합성 능력을 회복시키는 이차복귀돌연변이체의 발췌)

  • Ha, Hye-Jeong;Ryou, Sang-Mi;Lee, Kang-Seok;Jeon, Che-Ok
    • Microbiology and Biotechnology Letters
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    • v.39 no.1
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    • pp.93-96
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    • 2011
  • It has been shown that a nucleotide substitution at position 770 in Escherichia coli 16S rRNA, which is implicated in forming the evolutionary conserved B2c intersubunit bridge, has a detrimental effect on ribosome function. In order to isolate second-site revertants that complement ribosomes containing C770G, we performed a random mutagenesis of the 16S rRNA gene and selected clones that could produce more CAT protein translated by specialized ribosome. One of the clones contained two nucleotide substitutions at positions 569 and 904 (C569G and U904C) and these mutations partially complemented the loss of protein-synthesis ability caused by C770G. Further studies using the isolated revertant will provide information about which part of 16S rRNA is interacting with C770 and the consequence of the structure formed by these interactions in the process of protein synthesis.

Monitoring the Bacterial Community Dynamics in a Petroleum Refinery Wastewater Membrane Bioreactor Fed with a High Phenolic Load

  • Silva, Cynthia C.;Viero, Aline F.;Dias, Ana Carolina F.;Andreote, Fernando D.;Jesus, Ederson C.;De Paula, Sergio O.;Torres, Ana Paula R.;Santiago, Vania M.J.;Oliveira, Valeria M.
    • Journal of Microbiology and Biotechnology
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    • v.20 no.1
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    • pp.21-29
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    • 2010
  • The phenolic compounds are a major contaminant class often found in industrial wastewaters and the biological treatment is an alternative tool commonly employed for their removal. In this sense, monitoring microbial community dynamics is crucial for a successful wastewater treatment. This work aimed to monitor the structure and activity of the bacterial community during the operation of a laboratory-scale continuous submerged membrane bioreactor (SMBR), using PCR and RT-PCR followed by denaturing gradient gel electrophoresis (DGGE) and 16S rRNA libraries. Multivariate analyses carried out using DGGE profiles showed significant changes in the total and metabolically active dominant community members during the 4-week treatment period, explained mainly by phenol and ammonium input. Gene libraries were assembled using 16S rDNA and 16S rRNA PCR products from the fourth week of treatment. Sequencing and phylogenetic analyses of clones from the 16S rDNA library revealed a high diversity of taxa for the total bacterial community, with predominance of Thauera genus (ca. 50%). On the other hand, a lower diversity was found for metabolically active bacteria, which were mostly represented by members of Betaproteobacteria (Thauera and Comamonas), suggesting that these groups have a relevant role in the phenol degradation during the final phase of the SMBR operation.

Twenty-five unrecorded bacterial species of the Republic of Korea belonging to the phylum Actinomycetota discovered during surveys in 2021

  • Inhyup Kim;Wan-Taek Im;Kiseong Joh;Myung Kyum Kim;Jung-Hoon Yoon;Wonyong Kim;Taegun Seo
    • Journal of Species Research
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    • v.12 no.3
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    • pp.229-239
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    • 2023
  • We isolated and identified 25 unrecorded bacterial species belonging to the phylum Actinomycetota found in the Republic of Korea. Sequence comparison of 16S rRNA was performed using the NCBI BLAST and EzBioCloud database to identify 25 species, which had a 16S rRNA gene sequence similarity of >98.8% and were allocated as unrecorded species in the Republic of Korea. Among the 25 unrecorded bacterial strains, Streptomyces was the most common with nine species, followed by Leifsonia with two species. Isoptericola, Nocardioides, Dermacoccus, Sinomonas, Patulibacter, Marmoricola, Allobranchiibius, Aldersonia, Actinokineospora, Agromyces, Aeromicrobium, Cellulomonas, and Gordonia with one species each were also found. Twenty-five unrecorded species were excavated in various environments, such as tidal flats, ferns, soil, pine cones, moss, mud, wetlands, and plants. These isolates were characterized on the basis of their phylogenetic, biochemical properties, and morphological data, and species descriptions were provided.