• 제목/요약/키워드: 16S rRNA analysis

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서울 약수터의 지표세균 분포 및 16S rRNA 염기서열을 이용한 총대장균군 동정 및 계통분석 (Occurrence of Indicator Bacteria and Identification of Total Coliforms Using 16S rRNA Gene in Drinking Spring Water in Seoul)

  • 윤태호;이향;최금숙;이승주;이목영;어수미
    • 한국환경보건학회지
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    • 제39권6호
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    • pp.513-521
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    • 2013
  • Objectives: This study was performed in order to detect indicator bacteria in drinking spring water (DSW) samples in Seoul Metropolitan City, and to identify their genus through 16S rRNA sequencing and then assessing the genetic relation of their strains. Methods: For indicator bacteria detection and identification of total coliforms, we analyzed DSW between the spring and summer seasons. In particular, DSW samples were chosen from sites repeatedly found unsatisfactory in recent years. Results: Heterotrophic plate counts of DSW in the spring and summer season were investigated in the range of 0-550 and 0-800 CFU/mL, respectively. Total coliforms of these were 0-1,900 and 0-2,100 CFU/100mL, fecal coliforms were 0-600 and 0-550 CFU/100mL, and Escherichia coli were 0-7 and 0-326 MPN/100mL. The detection ratio of fecal pollution indicators and that of fecal coliforms increased to 58.6% in the summer from 12.5% in the spring and Escherichia coli increased to 51.4% from 4.7%. As a result of genetic analysis on the isolated bacteria, the genus of total coliforms was classified in the order of Enterobacter spp. 12.7%, Serratia spp. 7.3%, E. hermanii 6.4%, Rahnella spp. 5.5%, Hafnia spp. 4.5%, Escherichia coli 3.6%, Klebsiella spp. 3.6% in the spring season. In the summer season, it was classified in order of Klebsiella spp. 16.6%, Enterobacter spp. 13.0%, Escherichia coli 11.0%, Serratia spp. 8.6%, Raoultella spp. 7.0%, Kluyvera spp. 5.6% and Citrobacter spp. 3.0%. Conclusions: The increase of fecal pollution in summer indicates that special attention to drinking DSW is required.

가축사체 매몰지 토양의 미생물 군집 분석 (Analysis of Microbial Communities in Animal Carcass Disposal Soils)

  • 박정안;최낙철;김성배
    • 대한환경공학회지
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    • 제35권7호
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    • pp.503-508
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    • 2013
  • 본 연구의 목적은 가축사체 매몰지 토양의 침출수 오염에 따른 병원성 미생물에 의한 잠재적 위해성을 평가하기 위하여 미생물 군집을 조사하는 것이다. 경기도 지역에 위치한 가축사체 매몰지 세 군데(A, B, C) 토양을 대상으로 DNA를 추출하여, 16S rRNA 염기서열을 분석을 통해 미생물 군집을 조사하였다. 연구결과를 문(phylum)별로 구분해보면, A 토양은 전체 토양미생물이 Proteobacteria (100%) 1개의 문으로 동정되었으며, B 토양은 Actinobacteria (66.4%) > Proteobacteria (31.1%) > Bacteriodetes (2.1%) > Acidobacteria (0.3%) 순으로, C 토양은 Actinobacteria (63.1%) > Proteobacteria (36.9%) 순으로 분포하였다. 속(genus)별로 구분해보면, A 토양에서는 Pseudomonas가 98% 비율로 나타났고,B와 C 토양의 경우 Arthrobacter이 각각 68, 61%로 우점하였다. 세 군데(A, B, C) 토양 미생물 군집의 종 다양성을 Shannon 지수에 근거하여 분석한 결과, B 토양(3.45)과 C 토양(3.43)은 유사한 수준이었으나, A 토양(2.37)은 가장 낮게 계산되었다. 또한, 분석결과 Salmonella, Campylobacter 그리고 Clostridium perfringens과 같은 병원균도 발견되지 않았으나, 세균혈증을 일으키는 Ralstonia pickettii가 높은 농도로 관찰되었다. 본 연구에 사용된 가축 매몰지 토양은 침출수에 의한 미생물학적 오염도가 낮은 것으로 판단되지만, 가축매몰에 따른 병원성 미생물에 의한 토양의 잠재적 위해성을 평가하기 위하여 지속적인 모니터링이 필요하다.

Genetic Analysis of a Structural Motif Within the Conserved 530 Stem-Loop of Escherichia coli 16S rRNA

  • Szatkiewicz Jin P.;Cho Hyun-Dae;Ryou Sang-Mi;Kim Jong-Myung;Cunningham Philip R.;Lee Kang-Seok
    • Journal of Microbiology and Biotechnology
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    • 제16권4호
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    • pp.569-575
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    • 2006
  • The 530 stem-loop is a 46 nucleotide stem-loop structure found in all small-subunit ribosomal RNAs. Phylogenetic and mutational studies by others suggest the requirement for Watson-Crick interactions between the nucleotides 505-507 and 524-526 (530 pseudoknot), which are highly conserved. To examine the nature and functional significance of these interactions, a random mutagenesis experiment was conducted in which the nucleotides in the proposed pseudoknot were simultaneously mutated and functional mutants were selected and analyzed. Genetic analysis revealed that the particular nucleotide present at each position except 524 was not exclusively critical to the selection of functional mutants. It also indicated that basepairing interactions between the positions 505-507 and 524-526 were required for ribosomal function, and much weaker base-pairing interactions than those of the wild-type also allowed high ribosomal function. Our results support the hypothesis that the 530 pseudoknot structure may undergo a 'conformational switch' between folded and unfolded states during certain stages of the protein synthesis process by interacting with other ligands present in its environment.

A report of 5 unrecorded bacterial species of the Deinococcus genus in Korea

  • Lee, Jae-Jin;Kang, Myung-Suk;Joo, Eun Sun;Kim, Myung Kyum
    • Journal of Species Research
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    • 제5권1호
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    • pp.22-26
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    • 2016
  • Five bacterial strains designated DY37, BS333, JJ521, BM1, and DG13-2 were assigned to the genus Deinococcus were isolated from forest soil samples collected from Deogyusan, Busan, Changwon, and Seoul of South Korea. The isolates were Gram-staining negative or positive, and pale pink- or red-pigmented, short-rod shaped. Phylogenetic analysis based on 16S rRNA gene sequence revealed that strains DY37, BS333, JJ521, BM1, and DG13-2 were most closely related to Deinococcus aquatilis CCM $7524^T$ (with 99.0% similarity), D. ficus CC-FR2-$10^T$ (100.0%), D. grandis KS $0485^T$ (99.2%), D. roseus TDMA-$uv51^T$ (98.9%), and D. yunweiensis $YIM007^T$(100.0%), respectively. These 5 species have never been proposed in Korea; therefore 5 species of 1 genera in the family Deinococcaceae in the order Deinococcales within the class Deinococci are reported for proteobacterial species found in Korea.

Complete Mitochondrial Genome Sequences of Chinese Indigenous Sheep with Different Tail Types and an Analysis of Phylogenetic Evolution in Domestic Sheep

  • Fan, Hongying;Zhao, Fuping;Zhu, Caiye;Li, Fadi;Liu, Jidong;Zhang, Li;Wei, Caihong;Du, Lixin
    • Asian-Australasian Journal of Animal Sciences
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    • 제29권5호
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    • pp.631-639
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    • 2016
  • China has a long history of sheep (Ovis aries [O. aries]) breeding and an abundance of sheep genetic resources. Knowledge of the complete O. aries mitogenome should facilitate the study of the evolutionary history of the species. Therefore, the complete mitogenome of O. aries was sequenced and annotated. In order to characterize the mitogenomes of 3 Chinese sheep breeds (Altay sheep [AL], Shandong large-tailed sheep [SD], and small-tailed Hulun Buir sheep [sHL]), 19 sets of primers were employed to amplify contiguous, overlapping segments of the complete mitochondrial DNA (mtDNA) sequence of each breed. The sizes of the complete mitochondrial genomes of the sHL, AL, and SD breeds were 16,617 bp, 16,613 bp, and 16,613 bp, respectively. The mitochondrial genomes were deposited in the GenBank database with accession numbers KP702285 (AL sheep), KP981378 (SD sheep), and KP981380 (sHL sheep) respectively. The organization of the 3 analyzed sheep mitochondrial genomes was similar, with each consisting of 22 tRNA genes, 2 rRNA genes (12S rRNA and 16S rRNA), 13 protein-coding genes, and 1 control region (D-loop). The NADH dehydrogenase subunit 6 (ND6) and 8 tRNA genes were encoded on the light strand, whereas the rest of the mitochondrial genes were encoded on the heavy strand. The nucleotide skewness of the coding strands of the 3 analyzed mitogenomes was biased toward A and T. We constructed a phylogenetic tree using the complete mitogenomes of each type of sheep to allow us to understand the genetic relationships between Chinese breeds of O. aries and those developed and utilized in other countries. Our findings provide important information regarding the O. aries mitogenome and the evolutionary history of O. aries inside and outside China. In addition, our results provide a foundation for further exploration of the taxonomic status of O. aries.

Structure Analysis of 16S rDNA Sequences from Strains of Acidithiobacillus ferrooxidans

  • Peng, Hong;Yang, Yu;Li, Xuan;Qiu, Guanzhou;Liu, Xueduan;Huang, Jufang;Hu, Yuehua
    • BMB Reports
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    • 제39권2호
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    • pp.178-182
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    • 2006
  • Four strains of Acidithiobacillus ferrooxidans with different iron oxidation capacity were isolated from different mine drainage stations. The 16S rRNA gene of these strains were cloned and sequenced. Based on our sequences analysis on the four strain and the data on the other strains deposited in Genbank, all A. ferrooxidans may be classified into three phylogenetic groups. The analysis data showed that nucleotide variables (signature sites) were detected in 21 positions, and most of them were found in the first 800bp from 5' terminal except position 970 and 1375. Interestingly, the first 13 signature sites were located in two main regions:the first region (position 175-234) located in V2 while the second region (position 390-439) were detected in constant region between V2 and V3. Furthermore, the secondary structure and minimal free energy were determined in two regions among strains of three groups. These results may be useful in characterizing the microevolutionary mechanisms of species formation and monitoring in biohydrometallurgical application.

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

  • 이슬;김민석
    • 한국동물생명공학회지
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    • 제34권3호
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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.

A New Record of Juvenile Chromis mirationis (Perciformes: Pomacentridae) from Korea, Revealed by Molecular Analysis, with a Comparison to Juvenile Chromis notata

  • Song, Young Sun;Kwun, Hyuck Joon;Kim, Jin-Koo;Senou, Hiroshi
    • Fisheries and Aquatic Sciences
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    • 제17권2호
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    • pp.263-267
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    • 2014
  • A single juvenile pomacentrid specimen (5.9 mm standard length) was collected from the Korea Strait in October 2010. The specimen is characterized by punctate-stellate melanophores scattered on the operculum and dorso-ventral region in front of the caudal peduncle, the lack of melanophores on the posterior end of the anal fin base, the presence of 14 spines and 14 soft rays on the dorsal fin, and the presence of 2 spines and 12 soft rays on the anal fin. A molecular analysis based on mitochondrial DNA 16S rRNA sequences showed that this specimen is closely related to adult Chromis mirationis (d = 0.002), but that it differs from Chromis notata (d = 0.017). Juvenile C. mirationis differ from juvenile C. notata in having no melanophores on the posterior end of the anal fin base. We propose a new Korean name, "tti-ja-ri-dom" for C. mirationis.

Purification and Properties of Chitosanase from Chitinolytic $\beta$-Proteobacterium KNU3

  • Yi, Jae-Hyoung;Jang, Hong-Ki;Lee, Sang-Jae;Lee, Keun-Eok;Choi, Shin-Geon
    • Journal of Microbiology and Biotechnology
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    • 제14권2호
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    • pp.337-343
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    • 2004
  • A bacterial strain concurrently producing extracellular chitosanase and chitinase was isolated from soil and identified as a member of the $\beta$-subgroup of Proteobacteria through its 16S rRNA analysis and some biochemical analyses. The newly discovered strain, named as KNU3, had 99% homology of its 16S rRNA sequence with chitinolytic $\beta$-Proteobacterium CTE108. Strain KNU3 produced 34 kDa of chitosanase in addition to two chitinases of 68 kDa and 30 kDa, respectively. The purified chitosanase protein (ChoK) showed activity toward soluble, colloidal, and glycol chitosan, but did not exhibit any activity toward colloidal chitin. The optimum pH and temperature of ChoK were 6.0 and $70^{\circ}C$, respectively. The chitosanase was stable in the pH 4.0 to 8.0 range at $70^{\circ}C$, while enzyme activity was relatively stable at below $45^{\circ}C$. MALDI-TOF MS and N-terminal amino acid sequence analyses indicated that ChoK protein is related to chitosanases from Matsuebacter sp. and Sphingobacterium multivorum. HPLC analysis of chitosan lysates revealed that glucosamine tetramers and hexamers were the major products of hydrolysis.

Isolation and Characterization of Flavobacterium johnsoniae from Farmed Rainbow Trout Oncorhynchus mykiss

  • Suebsing, Rungkarn;Kim, Jeong-Ho
    • Fisheries and Aquatic Sciences
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    • 제15권1호
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    • pp.83-89
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    • 2012
  • Flavobacterium johnsoniae was isolated from farmed rainbow trout Oncorhynchus mykiss in Korea, and its biochemical and molecular characterization was determined. Yellow-pigmented bacterial colonies were isolated from 18 of 64 fish samples (28.1%) on trypticase soy agar plates, and their biochemical profiles were characterized by API 20E and API 20NE test kits. F. johnsoniae was identified by biochemical phenotyping of factors including rapid gliding motility, Gram-negative condition, oxidase- and catalase-positive status, Congo red absorption, nitrate reduction, ${\beta}$-galactosidase production, acid production from glucose, and gelatin and casein hydrolysis. PCR and subsequent sequencing of 16S rRNA confirmed that the yellow-pigmented colonies were most similar to F. johnsoniae. The alignment analysis of 16S rRNA sequences also showed that all 18 rainbow trout isolates had highly similar homologies (97-99% identity). One isolate was selected and named FjRt09. This isolate showed 98% homology with previously reported F. johnsoniae isolates, and in phylogenetic analysis was more closely grouped with F. johnsoniae than with F. psychrophilum, F. columnare, or F. branchiophilum. This is the first report on the occurrence and biochemical characterization of F. johnsoniae isolated from rainbow trout in Korea.