• Title/Summary/Keyword: 16S rRNA analysis

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Characterization of Miniimonas sp. S16 isolated from activated sludge (활성슬러지로부터 분리된 Miniimons sp. S16 세균의 특성)

  • Koh, Hyeon-Woo;Kim, Hongik;Park, Soo-Je
    • Korean Journal of Microbiology
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    • v.55 no.3
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    • pp.242-247
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    • 2019
  • Biological factors (e.g. microorganism activity) in wastewater treatment plant (WWTP) play essential roles for degradation and/or removal of organic matters. In this study, to understand the microbial functional roles in WWTP, we tried to isolate and characterize a bacterial strain from activated sludge sample. Strain S16 was isolated from the activated sludge of a municipal WWTP in Daejeon metropolitan city, the Republic of Korea. The cells were a Gram-stain-positive, non-motile, facultative anaerobe, and rod-shaped. Strain S16 grew at a temperature of $15{\sim}40^{\circ}C$ (optimum, $30^{\circ}C$), with 0~9.0% (w/v) NaCl (optimum, 1.0~2.0%), and at pH 5.5~9.0 (optimum, pH 7.0~7.5). Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain S16 was most closely related to the unique species Miniimonas arenae NBRC $106267^T$ (99.79%, 16S rRNA gene sequence similarity) of the genus Miniimonas. The cell wall contained alanine, glutamic acid, serine, and ornithine. Although the isolation source of the type strain NBRC $106267^T$ which considered as a marine microorganism is sea sand, that of strain S16 is terrestrial environment. It might raise an ecological question for habitat transition. Therefore, comparative genome analysis will be valuable investigation for shedding light on their potential metabolic traits and genomic streamlining.

Analysis of Plasmid pJP4 Horizontal Transfer and Its Impact on Bacterial Community Structure in Natural Soil

  • KIM TAE SUNG;KIM MI SOON;JUNG MEE KUM;JOE MIN JEONG;AHN JAE HYUNG;OH KYOUNG HEE;LEE MIN HYO;KIM MIN KYUN;KA JONG OK
    • Journal of Microbiology and Biotechnology
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    • v.15 no.2
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    • pp.376-383
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    • 2005
  • Alcaligenes sp. JMP228 carrying 2,4­dichlorophenoxyacetic acid (2,4-D) degradative plasmid pJP4 was inoculated into natural soil, and transfer of the plasmid pJP4 to indigenous soil bacteria was investigated with and without 2,4-D amendment. Plasmid pJP4 transfer was enhanced in the soils treated with 2,4-D, compared to the soils not amended with 2,4-D. Several different transconjugants were isolated from the soils treated with 2,4-D, while no indigenous transconjugants were obtained from the unamended soils. Inoculation of the soils with both the donor Alcaligenes sp. JMP228/pJP4 and a recipient Burkholderia cepacia DBO 1 produced less diverse transconjugants than the soils inoculated with the donor alone. Repetitive extragenic palindromic-polymerase chain reaction (REP-PCR) analysis of the transconjugants exhibited seven distinct genomic DNA fingerprints. Analysis of 16S rDNA sequences indicated that the transconjugants were related to members of the genera Burkholderia and Pandoraea. Denaturing gradient gel electrophoresis (DGGE) analysis of PCR-amplified 16S rRNA genes revealed that inoculation of the donor caused clear changes in the bacterial community structure of the 2,4-D­amended soils. The new 16S rRNA gene bands in the DGGE profile corresponded with the 16S rRNA genes of 2,4-D­degrading transconjugants isolated from the soil. The results indicate that introduction of the 2,4-D degradative plasmid as Alcaligenes sp. JMP228/pJP4 has a substantial impact on the bacterial community structure in the 2,4-D-amended soil.

Isolation of Cyanobacteria Producing Microcystin from Lakes (담수 생태계에서 Microcystin을 생산하는 남조세균의 분리)

  • Lee, Hee-Seon;Oh, Kyoung-Hee;Cho, Young-Cheol
    • Korean Journal of Microbiology
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    • v.44 no.3
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    • pp.251-257
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    • 2008
  • Four potential microcystin-producing cyanobacteria were isolated from large reservoirs which act as sources of drinking water supply in Korea. Strain DC-2, YD-l, and YD-6 were closely related to Microcystis aeruginosa based on the analysis of l6S rRNA gene and mcyA gene sequences. mcyA gene sequence of YDS2-3, isolated from Yongdam Reservoir, was closed to that of M. aeruginosa, whereas l6S rRNA gene sequence was not related to the known sequences of microcystin-producing cyanobacteria indicating this strain can be a novel cyanobacterium belonging to the genus Microcystis. When mcyA gene sequences of isolated cyanobacteria were compared with the mcyA gene sequence library of two reservoirs, the sequence of DC-2 matched with the dominant ones.

Analysis of Archaeal Communities in Full-Scale Anaerobic Digesters Using 454 Pyrosequencing (454 Pyrosequencing을 이용한 실규모 혐기성 소화조의 아케아 군집구조 분석)

  • Kang, Hyun-Jin;Kim, Taek-Seung;Lee, Young-Haeng;Lee, Taek-June;Han, Keum-Suk;Choi, Young-Jun;Park, Hee-Deung
    • Korean Journal of Microbiology
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    • v.47 no.3
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    • pp.209-217
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    • 2011
  • Archaeal communities were investigated using 454 pyrosequencing technology based on 16S rRNA gene in 11 samples collected from six different full-scale anaerobic digesters. Observed operational taxonomic units (OTUs) estimated from the archaeal 16S rRNA gene sequences were 13-55 OTUs (3% cutoff) which was corresponded to 29-89% of Chao1 richness estimates. In the anaerobic digesters there were archaeal sequences within the orders Thermoproteales, Thermoplasmatales, Desulfurococcales as well as within the orders Methanomicrobiales, Methanobacteriales, Methanococcales, Methanosarcinales, and Methanocellales, which are known to produce methane. Among these orders, Methanococcales known to produce methane using hydrogen was the predominant taxon and constituted 51.8-99.7% of total sequences. All samples showed a very similar community structure (Pearson correlation coefficient=0.99) except for one sample based on a heat map analysis. In addition, canonical correspondence analysis correlating archaeal communities to the environmental variables demonstrated that digester temperature and total solids removal rate were the two important explanatory variables. Overall results suggested that environmental and operational variables of anaerobic digester are important factors determining archaeal diversity and community structure.

Isolation and identification of the abundant bacteria in dental caries in children (소아의 치아우식 부위별 우점 세균 분리 및 동정)

  • Kim, Eun-Mi
    • Journal of Korean society of Dental Hygiene
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    • v.18 no.5
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    • pp.843-852
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    • 2018
  • Objectives: The study aimed to isolate the abundant bacteria in dental caries in children and to investigate the bacterial species involved in addition to those that have been previously reported. Methods: The specimens were collected from the supragingival plaques of each dental caries area, pit and fissure caries, deep dentinal caries, smooth surface caries, and dental caries, and from healthy subjects in the control group. Bacteria were cultured from these specimens, DNA was extracted from the isolated bacteria, and the 16S rRNA gene sequences were analyzed and identified. Results: Based on the results of the 16S rRNA gene sequence analysis for the 90 strains of dominant bacteria from the 45 specimens, 5, 7, 8, 7, and 13 species were identified from the supragingival plaques from healthy teeth, pit and fissure caries, deep dentinal caries, smooth surface caries, and dental caries, respectively. In healthy teeth, Actinomyces naeslundii dominated. Corynebacterium durum, Ralstonia pickettii, and Streptococcus intermedius showed equal distribution. The dominant bacterial species in dental caries, S. sanguinis, showed the greatest difference in prevalence in pit and fissure caries. In deep dentinal caries, S. mutans and Lactobacillus rhamnosus were dominant; in smooth surface caries, S. mutans and S. sanguinis were dominant; and in the supragingival plaques of dental caries, S. sanguinis and S. mutans were dominant. Conclusions: The bacterial species isolated from dental caries encompassed four phyla, eight genera, and 22 species. In addition, the SS1-2 strain, belonging to the genus Neisseria, was identified as a new species from among the isolated strains.

The Phylogenetic Relationship of Several Oscillatorian Cyanobacteria, Forming Blooms at Daecheong Reservoirs, Based on Partial 16S rRNA Gene Sequences

  • Lee, Wook-Jae;Bae, Kyung-Sook
    • Journal of Microbiology and Biotechnology
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    • v.11 no.3
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    • pp.504-507
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    • 2001
  • The partial 16S gene sequences of six filamentous cyanobacterial strains, Oscillatoria lmosa KCTC AG10168, Oscillatoria princeps KCTC AG10153, Oscillatoria sp. KCTC AG 10184, Phormidium tenue KCTC AG10158, Phormidium parchydematicum KCTC AG10164, and Lyngbya hieronymusii KCTC AG10199, which were isolated in the late summer at Daecheong Reservoirs, were determined and assigned their phylogenetic and taxonomic position among taxa of order Ocillatoriales whose partial 16S rRNA gene sequences aligned in this suty, were very heterogeneously clustered with other taxa. The two strains, Oscillatoria limosa KCTC AG10168 and O. princeps KCTC AG10153, formed a cluster with O. sancta PCC7515, which supported 64% of the bootstrap trees with high similarity (19-96.15%). Strain Oscillatoria sp. KCTC AG10184, that was known to produce a nasty substance, was closely related to the toxic Oscillatoria group. The study on morphological variation in various environments and toxin production will confirm the taxonomic status of these species. Phormidium tenus KCTC AG10158 and Phormidium parchydematicum KCTC AG10164 made a cluster with other oscillatorian species of Phormidium, Oscillatoria, and Leptolynbya, which supproted 100% of the bootstrap trees with a very high sequence smilarity (96.8-99.8%) in thsi study. The sequence analysis in this study also supported that taxa of oscillatoriales are not monophyletic. Some of the fractures, such as the presence or absence of sheath and cell shape, which were used to define them, would be inadequate and should be reconfirmed. We suggest that sequences of partial 16S rRNA gene fragments aligned in this study should be more useful than morphological features in the identification and reconfirmation of the taxonomic status of these oscillactorian cyanobacteria.

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Analysis of 16S-23S rRNA Intergenic Spacer Regions of Aeromonas veronii biogroup sobria and A. caviae (Aeromonas veronii biogroup sobria와 Aeromonas caviae의 16S-23S rRNA Intergenic Spacer Regions 분석)

  • 강동율;이훈구
    • Korean Journal of Microbiology
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    • v.36 no.3
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    • pp.173-180
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    • 2000
  • The intern1 spacer regions (ISR) between the 16s and 23s $1_RNA$ genes of Aeronzonus iwonii blogroupsobria and A. caviae were investigated by PCR fragment length typing and DNA sequencing. A. iwonii bv.sobria has a speciIic 16s-23s pattern of 2-4 fiagments ranging Goin 479-539 bp, with the exception of thespecies Aeron7onns cmiae, which has 3 fragments ranglog from 470-602 bp. In all of the.4 vei*onii bv. sobr,iaand A, caviae strains examined in this study, the 470-481bp Tragnent, designated TSR-1, invariably contained $tDNA^{uc(GAT)$ and $tDNA^{Ala(TGC)$ in contrast to ISR-2 (513-525 bp). ISR-3 (537-539 bp) and ISR-4 (568-602 bp)containing TEX>$tDNA^{Olu(ITC)$ A stretch of 20 nucleotides (178-197 bp) in the ISR-4 was conserved only wit11mA.caiiue, from which the A. caiiae specific primer, named prAC-F, was designed and used for PCR with aAcaviae coimnon reverse primer A PCR product of 450 bp was apparent alnong I , caiizne strains, but not ii1.4.ijeronii bv. sob~ia strains. The PCR product was oot detected t"-om strains belonging to A. hjili-o~~hila, Ebrio,aud the family Ef\ulcornertei,obncteriaceae. This study provides the first molecular tool for mdentifying the species 8.caviae.ing the species 8. caviae.

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Comparison of Culture-dependent and DGGE based Method for the Analysis of Marine Bacterial Community (배양법과 DGGE에 의한 해양세균 군집의 비교분석)

  • Kim, Mal-Nam;Bang, Hyo-Joo
    • Korean Journal of Environmental Biology
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    • v.24 no.4
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    • pp.307-313
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    • 2006
  • Seasonal variation of marine bacterial community was analyzed in the surface sea water collected from one of the stations locating at Tongyeoung coastal area, Korea. The results obtained by the culture method through identification with the VITEK Microbe ID system after pure culture in the selective medium were compared with those obtained by the DGGE based 16S rRNA PCR method. The composition of the marine bacterial community in the sea water samples harvested in September, 2004, November, 2004, January, 2005, May, 2005 and August, 2005 determined by the culture method showed 5, 5, 4, 6, and 10 strains respectively. Pseudomonas fluorescens and Acinetobacter lwoffii were detected in all seasons. The other strains were identified to be Pseudomonas stutzeri, Sphingomonas paucimobilis, Burkholderia mallei and Chryseobacterium indologenes. In contrast, the 16S rRNA PCR-DGGE method detected 10, 11, 6, 9 and 13 populations respectively in the same sea water samples and the strains were identified to be Acinetobacter lwoffii, Burkholderia mallei, Pseudomonas fluoresence, Actinobacillus ureae, Burkholderia sp., Pseudomonas stutzeri, Roseobacter sp., Vibrio parahaemolyticue, Sphingomonas paucimobilis and Rugeria algocolus. This results indicated that the DGGE based 16S rRNA PCR method was more efficient than the culture method for the grasp of the characteristics of the marine bacterial community.

Molecular Identification of the Bloom-forming Cyanobacterium Anabaena from North Han River System in Summer 2012 (북한강 수계 조류대발생 원인종 남조 Anabaena의 분자계통학적 검토)

  • Li, Zhun;Han, Myung-Soo;Hwang, Su-Ok;Byeon, Myeong-Seop;Hwang, Soon-Jin;Kim, Baik-Ho
    • Korean Journal of Ecology and Environment
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    • v.46 no.2
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    • pp.301-309
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    • 2013
  • Between May and August 2012, a massive cyanobacterial bloom with Anabaena has been occurred throughout the North Han River. Sampling was conducted at one station on each lake, L. Uham, L. Cheongpyung, and L. Paldang, where occurred a dense bloom, in 13 July. According to the microscopic examination, the blooms was dominated by one specific filamentous cyanobacterium Anabaena and other phytoplankton. Morphologically, previous literature proven that this Anabaena species is A. crassa (Lemmermann) Komark.-Legn. & Cronberg. However, identification of species in a mixed population is complicated due to limited morphological differences. Therefore, with live sample including trichome, akinete and heterocyst, the sequences of 16S rRNA gene of Anabaena isolates were cloned and analyzed, and three 16S rRNA gene sequences of 1188~1520 bp in length were obtained. It was shown from the homologous analysis results that the obtained 16S rRNA sequences were highly homologous to the relevant sequences of A. crassa in GenBank. The 16S rRNA sequences of 63 species were retrieved from GenBank, and the phylogenetic tree was constructed by using these sequences.

Studies on the mycoplasmal pneumonia in slaughter pigs. 2. Isolation of mycoplasmas from lung tissues and identification of isolates by nested-PCR technique (도축돈의 마이코플라즈마성 폐렴에 관한 연구 2. 폐조직에서의 균분리와 nested-PCR방법에 의한 동정)

  • Lim, Young-Taek;Seok, Ho-Bong
    • Korean Journal of Veterinary Research
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    • v.42 no.2
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    • pp.225-229
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    • 2002
  • We report that mycoplasma organisms from lung tissues of slaughter pigs were identified to genes fragments with references use of nested-PCR technique(nPCR). Seven strains of mycoplasma species were isolated from 70 lung tissues. The organisms were detected by in vitro amplification of 16S rRNA and 23S rRNA genes. Nucleotide sequences of the spacer between 16S and 23S in the ribosomal RNA operons of mycoplasma were identified by the analysis of products from the nested PCR. Four common PCR primers, MhF1, MhF2 MhR1 and MhR2, were designed by analysis between these sequences by first amplified with F1, R1 and second with F2, R2, respectively. Specific amplification of the spacer region for reference strains of M. hyopneumoniae, M. hyorhinis, M. flocculare were confirmed by first round of PCR in which the traduced fragments of 690bp, 460bp, 630bp. But amplications of second round was changed to 240bp, 210bp, 230bp, respectively. Three different strains (M. hyopneumoniae:4, M. hyorhinis:2, M. flocculare:1) were detected by the nested-PCR technique. The results suggest that the detection of swine mycoplasma by n-PCR can be analyzed the nucleotide sequences between rRNA operons and homology study.