• Title/Summary/Keyword: 16S rRNA gene sequences

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Identification and Distribution of Bacillus Species in Doenjang by Whole-Cell Protein Patterns and 16S rRNA Gene Sequence Analysis

  • Kim, Tae-Woon;Kim, Young-Hoon;Kim, Sung-Eon;Lee, Jun-Hwa;Park, Cheon-Seok;Kim, Hae-Yeong
    • Journal of Microbiology and Biotechnology
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    • v.20 no.8
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    • pp.1210-1214
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    • 2010
  • Many bacteria are involved in the fermentation of doenjang, and Bacillus species are known to perform significant roles. Although SDS-PAGE has been frequently used to classify and identify bacteria in various samples, the microbial diversity in doenjang has not yet been investigated. This study aims to determine the identity and distribution of dominant Bacillus species in doenjang using SDS-PAGE profiles of whole-cell proteins and 16S rRNA gene sequencing. Reference Bacillus strains yielded differential SDS-PAGE banding patterns that could be considered to be highly specific fingerprints. Grouping of bacterial strains isolated from doenjang samples by whole-cell protein patterns was confirmed by analysis of their 16S rRNA gene sequences. B. subtilis was found to be the most dominant strain in most of the samples, whereas B. licheniformis and B. amyloliquefaciens were less frequently found but were also detected in several samples. The results obtained in this study show that a combined identification method using SDS-PAGE profiles of whole-cell proteins and subsequent 16S rRNA gene sequence analysis could successfully identify Bacillus species isolated from doenjang.

Six newly recorded species of epilithic cyanobacteria isolated in Korea

  • Hye-Ryeung Wang;Ji-Ho Song;Nam-Ju Lee;Do-Hyun Kim;So-Won Kim;Ok-Min Lee
    • Journal of Species Research
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    • v.13 no.1
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    • pp.10-31
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    • 2024
  • In this study, 11 strains of epilithic cyanobacteria belonging to six unrecorded species in Korea were isolated from gravel submerged in freshwater of seven collection sites in Korea. The morphological characteristics of the six species isolated in this study were consistent with the type strain of each species, and the similarity of the 16S rRNA gene sequences with the type strain of each species were 98.8-100%. In the phylogenetic tree using the 16S rRNA gene sequences, the 11 strains of these six species formed the same cluster as the strains of each species. The habitat of each previously reported species is mainly the soil surface, but all Korean strains appeared from the gravel submerged in freshwater. As a result of the morphological, ecological, and molecular analyses, these six species of cyanobacteria were identified as Geminocystis papuanica, Allocoleopsis franciscana, Ancylothrix terrestris, Klisinema persicum, Scytolyngbya timoleontis, and Shackletoniella antarctica, which were added as newly recorded species in Korea.

Identification and Comparison of the Nucleotide Sequence of 16S-23S rRNA Gene Intergenic Small SR(Spacer Region) of Lactobacillus rhamnosus ATCC 53103 with Those of L. casei, L. acidophilus and L. helveticus

  • Byun, J.R.;Yoon, Y.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.12
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    • pp.1816-1821
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    • 2003
  • Reliable PCR based identification of lactobacilli has been described utilizing the sequence of 16S-23S rRNA intergenic spacer region. Those sequence comparisons showed a high degree of difference in homology among the strains of L. rhamnosus, L. casei, L. acidophilus and L. helveticus whose 16S-23S rRNA intergenic small SR's sizes were 222 bp, 222 bp, 206 bp and 216 bp respectively. The sequence of 16S-23S rRNA intergenic spacer region of L. rhamnosus ATCC 53103 revealed the close relatedness to those of L. casei strains by the homology ranges from 95.4% to 97.2%. 16S-23S rRNA intergenic spacer region nucleotide sequence of L. acidophilus showed some distant relatedness with L. rhamnosus ATCC 53103 with the homology ranges from 40.3% to 41.8% and that with L. helveticus was shown to be 30% of homology, which exists at the most distant phylogenetic relatedness. The identification of species and strain of lactobacilli was possible on the basis of these results. The common sequences among the 17 strains were CTAAGGAA located in the initiating position of the DNA and some discrepancies were found between the same strains based on these results.

Diversity Census of Fecal Microbiome in Horses (말 분변 내 마이크로바이옴 다양성 조사)

  • Lee, Seul;Kim, Minseok
    • Journal of Animal Reproduction and Biotechnology
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    • v.34 no.3
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    • pp.157-165
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    • 2019
  • This study was conducted to analyze the diversity census of fecal microbiome in horses using meta-analysis of equine 16S rRNA gene sequences that are available in the Ribosomal Database Project (RDP; Release 11, Update 5). The search terms used were "horse feces (or faeces)" and "equine feces (or faeces)". A total of 842 sequences of equine feces origin were retrieved from the RDP database, where 744 sequences were assigned to 10 phyla placed within Domain Bacteria. Firmicutes (n = 391) and Bacteroidetes (n = 203) were the first and the second dominant phyla, respectively, followed by Verrucomicrobia (n = 58), Proteobacteria (n = 30) and Fibrobacteres (n = 24). Clostridia (n = 319) was the first dominant class placed within Bacteroidetes while Bacteroidia (n = 174) was the second dominant class placed within Bacteroidetes. The remaining 98 sequences were assigned to phylum Euryarchaeota placed within Domain Archaea, where 74 sequences were assigned to class Methanomicrobia. The current results will improve understanding of the diversity of fecal microbiome in horses and may be used to further analyze equine fecal microbiome in future studies.

Genetic Phylogeny among Three Species Red Seabream, Black Seabream and Rock Bream Based on Mitochondrial DNA Sequences

  • Kim, Mi-Jung;An, Hye-Suck;Kim, Kyung-Kil;Park, Jung-Youn
    • Fisheries and Aquatic Sciences
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    • v.12 no.3
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    • pp.171-178
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    • 2009
  • The Perciformes include approximately 40% of all bony fishes and are the largest order of vertebrates. This order includes some of the most economically relevant marine fishes, particularly the red seabream, black seabream and rock bream. A 409 bp fragment of the cytochrome b (cyt b) gene and 403 bp and 518 bp fragments of ribosomal RNA (12S and 16S rRNA, respectively) were sequenced from five populations of natural and cultured red seabreams, natural black seabream, and natural and cultured rock breams. The mitochondrial DNA sequences were utilized for the genetic identification and population structural analyses of these three species. Phylogenetic relationships of intra- and inter-species were elucidated using three types of molecular genetic markers from three species of the order Perciformes in Korea. We noted no significant differences in the intra-specific variation of the cyt b and rRNA genes in each population however, inter-specific divergences were greater than intra-specific variation. Inter-specific variation was induced more by transition than transversion type in the cyt b and rRNA genes. The cyt b gene and rRNA genes make it possible to determine the inter-species divergence. The rRNA genes have more conserved sequences than the cyt b gene. Therefore, these genes are expected to prove useful among species belonging to the different genera or families.

Phylogenetic Analysis of Pectobacterium Species Using the 16S-23S rRNA Intergenic Spacer Regions

  • Kwon, Soon-Wo;Cheun, Meung-Sook;Kim, Sang-Hee;Lim, Chun-Keun
    • The Plant Pathology Journal
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    • v.16 no.2
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    • pp.98-104
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    • 2000
  • For the taxonomic evaluaition, 15 strains of the genus Pectobacterium and Erwinia were analyzed for 16S-23S rDNA intergenic spacer regions (ISRs). These species contained two types of ISRs, large and small ISRs. Large ISRs were on the range of 474-569 bp size, and coding transfer $\textrm{RNA}^{11e}$($\textrm{tRNA}^{11e}$) and $\textrm{tRNA}^{Ala}$. Small ISRs were 354-459 bp in length and coding $\textrm{tRNA}^{Glu}$. The sequence variations of two ISRs among species and strains were very high as compared with 16S rRNA gene sequences. By phylogenetic trees on the basis of two ISRs, Pectobacterium ere differentiated into P. carotovorum-P. cactiaidum group and P. chrysanthemi group. However, the taxonomic position of E. cypripedii and E. rhapontici, which were not clear on taxonomic delineation between Pectobacterium and Erwinia, were not clearly resolved on the basis of ISRs.

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16S rRNA gene-based sequencing of cucumber (Cucumis sativus L.) microbiota cultivated in South Korea (16S rRNA 유전자 염기서열 분석에 기반한 국내 재배 오이의 상재균총 분석)

  • Seo, Dong Woo;Kim, Seung Min;Lee, Heoun Reoul;Yum, Su-jin;Jeong, Hee Gon
    • Korean Journal of Food Science and Technology
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    • v.53 no.3
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    • pp.334-343
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    • 2021
  • Various vegetables, including cucumbers, have a high probability of foodborne illness because they are usually eaten raw. In this study, we analyzed the 16S rRNA gene sequences of the cucumber (Cucumis sativus L.) microbiota. The diversity indices of cucumber cultivated in May were higher than in cucumber cultivated in November. At the phylum level, Proteobacteria, Firmicutes, and Actinobacteria were predominant. The classes generally comprised Gammaproteobacteria, Bacilli, Alphaproteobacteria, and Actinobacteria. At the genus level, the proportions of Aureimonas, Escherichia, and Microbacterium in samples from May were relatively high, whereas Enterococcus, Pseudomonas, and Rhizobium accounted for a higher proportion in samples from November. Moreover, it is noteworthy that potential pathogenic genera such as Acinetobacter, Aerococcus, Aureimonas, Enterobacter, Enterococcus, Escherichia, Pantoea, Pseudomonas, and Staphylococcus were detected. Although further studies on the characteristics of potential pathogens are required, our results can be used to improve the food safety of vegetables.

Genetic identification of Aeromonas species using a housekeeping gene, rpoD, in cultured salmonid fishes in Gangwon-Do (강원도 양식 연어과 어류에서 분리된 에로모나스 종의 유전학적 동정)

  • Lim, Jongwon;Koo, Bonhyeong;Kim, Kwang Il;Jeong, Hyun Do;Hong, Suhee
    • Journal of fish pathology
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    • v.30 no.2
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    • pp.79-88
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    • 2017
  • At the present, fish farms are suffering a lot of economic losses due to infectious diseases caused by various pathogens including aeromonad. Aeromonad is ubiquitous bacteria that causes infectious diseases. At least 26 species in the genus Aeromonas have been reported to cause fatal infections not only in salmonid fishes, but also in other freshwater and seawater fishes. Molecular techniques based on nucleic acid sequences of 16S rDNA and housekeeping genes can be used to identify the Aeromonas species. In this study, The genus Aeromonas was isolated from salmonid fishes of sixteen fish farms in Gangwon-Do, Korea and phylogenetically identified based on the sequences of 16S rDNA and housekeeping genes for Aeromonad, i.e. RNA polymerase sigma factor ${\sigma}^{70}$ (rpoD) or DNA gyrase subunit B (gyrB). Consequently, 96 strains were collected from Atlantic salmon (Salmo salar), coho salmon (Oncorhynchus kisutch), masou salmon (Oncorhynchus masou) and rainbow trout (Oncorhynchus mykiss), and 36 isolates were identified as the genus Aeromonas by 16S rDNA analysis. Thirty six Aeromonad isolates were further analysed based on rpoD or gyrB gene sequences and found Aeromonas salmonicida (24 isolates), A. sobria (10 isolates), A. media (1 isolates) and A. popoffii (1 isolates), indicating that A. salmonicida is a main infectious bacteria in Salmonid fishes in Gangwon-Do. It was also proved that the phylogenetic identification of Aeromonas species based on the sequences of housekeeping gene is more precise than the 16S rDNA sequence.

A report of 20 unrecorded bacterial species in Korea, isolated from soils of coastal areas in 2022

  • Seung Hyeok Soung;Jaeho Song;Seung Yeol Shin;Song-Ih Han
    • Journal of Species Research
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    • v.12 no.4
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    • pp.267-276
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    • 2023
  • To obtain unrecorded bacterial species in Korea, various soils of coastal areas were collected from the Republic of Korea in 2022. After plating the samples on marine agar and incubating aerobically and anaerobically, approximately 1,700 bacterial strains were isolated and identified using 16S rRNA gene sequences. A total of 20 strains showed ≥98.7% 16S rRNA gene sequence similarity with validly published bacterial species but not reported in Korea, indicating they are unrecorded bacterial species in Korea. The unrecorded bacterial strains belonged to four phyla, six classes, 15 orders, 16 families, and 19 genera which were assigned to Blastomonas and Sphingomonas of the class Alphaproteobacteria; Pseudidiomarina, Kushneria, Salinicola, and Salinisphaera of the class Gammaproteobacteria; Evansella, Virgibacillus, and Paenibacillus of the class Bacilli; Cyclobacterium of the class Cytophagia; Pedobacter of the class Sphingobacteriia; and Demequina, Ornithinimicrobium, Blastococcus, Jatrophihabitans, Kineococcus, Glaciihabitans, Aeromicrobium and Streptomyces of the class Actinomycetes. The details of the 20 unreported species, including Gram reaction, morphology, biochemical characteristics, and phylogenetic position are also provided in the description of the strains.