• Title/Summary/Keyword: 16S-rRNA

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Microbial Community Analysis using RDP II (Ribosomal Database Project II):Methods, Tools and New Advances

  • Cardenas, Erick;Cole, James R.;Tiedje, James M.;Park, Joon-Hong
    • Environmental Engineering Research
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    • v.14 no.1
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    • pp.3-9
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    • 2009
  • Microorganisms play an important role in the geochemical cycles, industry, environmental cleanup, and biotechnology among other fields. Given the high microbial diversity, identification of the microorganism is essential in understanding and managing the processes. One of the most popular and powerful method for microbial identification is comparative 16S rRNA gene analysis. Due to the highly conserved nature of this essential gene, sequencing and later comparison of it against known rRNA databases can provide assignment of the bacteria into the taxonomy, and the identity of its closest relatives. Isolation and sequencing of 16S rRNA genes directly from natural environments (either from DNA or RNA) can also be used to study the structure of the whole microbial community. Nowadays, novel sequencing technologies with massive outputs are giving researchers worldwide the chance to study the microbial world with a depth that was previously too expensive to achieve. In this article we describe commonly used research approaches for the study of individual microorganisms and microbial communities using the tools provided by Ribosomal Database Project website.

Molecular Phylogeny of the Family Tephritidae (Insecta: Diptera): New Insight from Combined Analysis of the Mitochondrial 12S, 16S, and COII Genes

  • Han, Ho-Yeon;Ro, Kyung-Eui
    • Molecules and Cells
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    • v.27 no.1
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    • pp.55-66
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    • 2009
  • The phylogeny of the family Tephritidae (Diptera: Tephritidae) was reconstructed from mitochondrial 12S, 16S, and COII gene fragments using 87 species, including 79 tephritid and 8 outgroup species. Minimum evolution and Bayesian trees suggested the following phylogenetic relationships: (1) A sister group relationship between Ortalotrypeta and Tachinisca, and their basal phylogenetic position within Tephritidae; (2) a sister group relationship between the tribe Acanthonevrini and Phytalmiini; (3) monophyly of Plioreocepta, Taomyia and an undescribed new genus, and their sister group relationship with the subfamily Tephritinae; (4) a possible sister group relationship of Cephalophysa and Adramini; and (5) reconfirmation of monophyly for Trypetini, Carpomyini, Tephritinae, and Dacinae. The combination of 12S, 16S, and COII data enabled resolution of phylogenetic relationships among the higher taxa of Tephritidae.

Analysis of Microbial Diversity in Nuruk Using PCR-DGGE (PCR-DGGE를 이용한 누룩에서의 미생물 다양성 분석)

  • Kwon, Seung-Jik;Sohn, Jae-Hak
    • Journal of Life Science
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    • v.22 no.1
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    • pp.110-116
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    • 2012
  • Nuruk plays a significant role in the flavor and quality of Takju and Yakju, which are produced through saccharification and alcohol fermentation by various microorganisms. In this study, we identified microbial strains isolated from a plate count and PCR-denaturing gradient gel electrophoresis (DGGE) analysis targeting the 16S and 28S rRNA genes, in order to characterize bacterial and fungal diversity in Sansung Nuruk. The numbers of bacteria and fungi in Nuruk were $1.5{\times}10^9$ CFU/g and $2.2{\tims}10^8$ CFU/g, respectively. The 16S rRNA gene sequence indicated that the predominant bacteria in the isolates and PCR-DGGE profile of Nuruk were Kocuria spp., Pantoea spp., Lactobacillus spp., Pediococcus spp., Weissella spp., Staphylococcus spp., endophytic bacterium, uncultured Gamma-proteobacteria, uncultured Cyanobacteria, and Actinobacteria. Dominant bacteria from the PCR-DGGE profile were Pediococcous pentosaceus and uncultured Cyanobacteria. The 28S rRNA gene sequence indicated the predominant fungi in the isolates and PCR-DGGE profile to be Trichomonascus spp. Pichia spp., Torulaspora spp., Wickerhamomyces spp., Sacharomycopsis spp., Lichtheimia spp., Mucor spp., Rhizopus spp. Aspergillus spp., and Cladosporium spp. Dominant fungi from the PCR-DGGE profile were Pichia kudriavzevii and Aspergillus oryzae. The PCR-DGGE technique was used for the first time in this study to assess a microbial community in Nuruk and proved to be an effective protocol for profiling microbial diversity.

Cloning and Sequencing of Resistance Determinants to Aminoglycoside Antibiotics from Sterptoalloteichus hindustanus ATCC 31219 (Streptoalloteichus hindustanus ATCC 31219로부터 아미노글라이코사이드계 항생제에 내성을 지정하는 유전자의 클로닝 및 염기서열 결정)

  • Kim, Jong-Woo;Han, Jae-Jin;Choi, Young-Nae;Eom, Joon-Ho;Yoon, Sung-Joon;Hyun, Chang-Gu;Suh, Joo-Won
    • Microbiology and Biotechnology Letters
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    • v.23 no.4
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    • pp.384-389
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    • 1995
  • Streptoalloteichus hindustanus ATCC 31219, a nebramycin complex producer, is similar to Streptomyeces tenebrarius in a viewpoint of resistance to a wide range of aminoglycoside antibiotics. S. tenebrarius has resistance mechanisms of 16s rRNA methylation and aminogycoside modification. However, it is not known whether resistance mechanisms of Stall. hindustanus are the same as in S. tenebrarius. Therefore, we have tried to isolate resistance determinants from Stall. hindustanus. Two different types of aminoglycoside resistance determinants were isolated from Stall. hindustanus and expressed in Streptomyces lividans TK24. The apramycin resistance gene (amr) and the tobramycin resistance gene (tmr) isolated from Stall. hindustanus showed broad resistance spectrum against a dozen of aminoglycoside antibiotics. The complete nucleotide sequences of apramycin resistance gene (amr) were determined. The deduced amino acid sequence of the amr gene of Stall hindustanus ATCC 31219 showed extensive sequence homology to the 16s rRNA methylase gene (kamB) of S. tenebrarius.

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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.

Characterization of Photobacterium damselae subsp. piscicida isolated from cultured starry flounder, Platichthys stellatus in Korea (우리나라 양식 강도다리, Platichthys stellatus에서 분리된 Photobacterium damselae subsp. piscicida의 특성)

  • Cho, Young Ah;Han, Hyun-Ja;Mun, Hee Eun;Jung, Sung Hee;Park, Myoung Ae;Kim, Jin Woo
    • Journal of fish pathology
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    • v.26 no.2
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    • pp.77-88
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    • 2013
  • Starry flounder, Platichthys stellatus (body length $4.4{\pm}0.51cm$) that became sick during an outbreak of disease at mariculture facilities at Ulsan, Korea in August of 2012, were examined to identify the cause of the disease. Diseased fish didn't show a unique sign, but the oxidase-positive and gram negative rod was isolated from moribund fish. The bacterium was revealed as Photobacterium damselae subsp. piscicida by biochemical analysis and sequence analysis of the 16S rRNA and capsular polysaccharide (CPS) genes. The isolates (AD5) was carrying susceptible to ofloxacin and gentamycin and showed high growth value at $18^{\circ}C$ and $25^{\circ}C$ compared to four other P. damsela strains.

Fibrinolytic Activity and Characterization of Bacillus licheniformis HK-12 Isolated from Chungkook-Jang (청국장에서 분리한 세균인 Bacillus licheniformis HK-12의 혈전용해활성 및 특징)

  • Sohn, Byung-Hee;Song, Yu-Jin;Oh, Kye-Heon
    • KSBB Journal
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    • v.23 no.3
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    • pp.251-256
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    • 2008
  • The aim of this work was to investigate the fibrinolytic activity and characterization of Bacillus licheniformis HK-12, which produces the fibrinolytic enzyme excreted from naturally fermented Chungkook-Jang. Initially, the physiological and biochemical characteristics of strain HK-12 was examined. Both physiological analysis using BIOLOG system and phylogenetic analysis using 16S rRNA sequencing were performed to identify the strain, and the strain could be assigned to Bacillus licheniformis, designated as B. lichenformis HK-12, and registered in GenBank as [EU288193]. Phylogenetic analysis of B. licheniformis HK-12 was plotted based on 16S rRNA sequence comparisons. During the incubation period of B. licheniformis HK-12, the changes of bacterial growth, fibrinolytic activity, and pH were monitored. As the results, after 36 hours of incubation, the maximum fibinolytic activity was about 2.25 times than that of plasmin used as standard. Optimal conditions on the growth of B. licheniformis HK-12 associated with the fibrinolytic activity was initial pH 7.0 and 40$^{\circ}C$, respectively.

Isolation and Identification of Thermostable \beta-glycosidase-producing Microorganism from Hot Spring of Volcanic Area at Atagawa in Japan. (일본의 Atagawa 온천지대에서 분리한 내열성 \beta-glycosidase 생성균주의 분리 및 동정)

  • 남은숙;최종우;차성관;안종건
    • Microbiology and Biotechnology Letters
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    • v.30 no.2
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    • pp.151-156
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    • 2002
  • This study was performed to obtain the thermostable $\beta$-glycosidase producing bacteria from hot spring of volcanic area at Atagawa in Japan. KNOUC 202 was selected because it showed thermostable $\beta$-glycosidase activity in sodium phosphate buffer(pH 6.8) at $70^{\circ}C$ for 4h, and it was identified. The strain was aerobic, asporogenic bacilli, immobile, gram negative, catalase positive, oxidase positive, and pigment-producing. Optimum growth was at $70~72^{\circ}C$, pH 7.0~7.2, and it could grow in the presence of 3% NaCl. The main fatty acids in cell were iso-15:0 and iso-l7:0. 16S rRNA sequence of KNOUC 202 showed 99.9% similarity with that of Thermus thermophilus ATCC 27634(HB8). Based on morphological, physiological, biochemical characteristics, cellular fatty acids profile and 16S rRNA sequence analysis, KNOUC 202 was identified as Thermus thermophilus.

Community Structure, Diversity, and Vertical Distribution of Archaea Revealed by 16S rRNA Gene Analysis in the Deep Sea Sediment of the Ulleung Basin, East Sea (16S rRNA 유전자 분석방법을 이용한 동해 울릉분지 심해 퇴적물 내 고세균 군집 구조 및 다양성의 수직분포 특성연구)

  • Kim, Bo-Bae;Cho, Hye-Youn;Hyun, Jung-Ho
    • Ocean and Polar Research
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    • v.32 no.3
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    • pp.309-319
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    • 2010
  • To assess community structure and diversity of archaea, a clone sequencing analysis based on an archaeal 16S rRNA gene was conducted at three sediment depths of the continental slope and Ulleung Basin in the East Sea. A total of 311 and 342 clones were sequenced at the slope and basin sites, respectively. Marine Group I, which is known as the ammonia oxidizers, appeared to predominate in the surface sediment of both sites (97.3% at slope, 88.5% at basin). In the anoxic subsurface sediment of the slope and basin, the predominant archaeal group differed noticeably. Marine Benthic Group B dominated in the subsurface sediment of the slope. Marine Benthic Group D and Miscellaneous Crenarchaeotal Group were the second largest archaeal group at 8-9 cm and 18-19 cm depth, respectively. Marine Benthic Group C of Crenarchaeota occupied the highest proportion by accounting for more than 60% of total clones in the subsurface sediments of the basin site. While archaeal groups that use metal oxide as an electron acceptor were relatively more abundant at the basin sites with manganese (Mn) oxide-enriched surface sediment, archaeal groups related to the sulfur cycle were more abundant in the sulfidogenic sediments of the slope. Overall results indicate that archaeal communities in the Ulleung Basin show clear spatial variation with depth and sites according to geochemical properties the sediment. Archaeal communities also seem to play a significant role in the biogeochemical carbon (C), nitrogen (N), sulfur (S), and metal cycles at each 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.