• Title/Summary/Keyword: 16S rDNA 염기서열

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Nucleotide Sequence of an Extracellular Phospholipase D Gene from Streptomyces somaliensis and Transphosphatidylation Activity of Its Enzyme (Streptomyces somaliensis가 생산하는 세포외 Phospholipase D의 유전자 서열 분석과 Transphosphatidylation 활성 특성)

  • Jeong Sujin;Lee Sun-Hee;Uhm Tai-Boong
    • Korean Journal of Microbiology
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    • v.40 no.3
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    • pp.211-216
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    • 2004
  • A bacterial strain JE-ll found to produce active extracellular phospholipase D (PLD) was selected from the soil isolates. It was identified as Streptomyces somaliensis on the basis of 16S rDNA sequence analysis, morphological and physiological characteristics. The gene (sspld) encoding S. somaliensis PLD was isolated and characterized. The open reading frame was suggested to encode 538 amino acids with a signal peptide of 33 amino acids. The deduced amino acid sequence of the sspld shared a sequence similarity of 70-88% with PLDs of other Streptomyces sp. so far reported. The PLD converted phosphatidylcholine to phosphatidylglycerol or phosphatidylserine with the yield of 96 to 99% (㏖/㏖), but did not act on inositol or ethanolamine as a transphosphatidylation donor.

Divergence Analysis of 16S rRNA and rpoB Gene Sequences Revealed from the Harmful Cyanobacterium Microcystis aeruginosa (유해 남조세균 Microcystis aeruginosa의 16S rRNA 및 rpoB 유전자 염기서열 변이 분석)

  • Ki, Jang-Seu
    • Korean Journal of Microbiology
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    • v.46 no.3
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    • pp.296-302
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    • 2010
  • Microcystis (Cyanobacteria, Chroococcales) is one of the green tide-causing organisms in freshwaters, and some species produce microcystin that is hepatotoxin. In the aspects of freshwater quality controls and health concerns, therefore it is necessary to manage the harmful organisms. In the present study, RNA polymerase beta subunit (rpoB) gene sequences of Microcystis were determined and characterized in order to use a potential marker for the molecular detections of the species. Microcystis rpoB showed high divergences of DNA similarity and genetic distances when compared with those of 16S rRNA, and the molecular differences were statistically significant (Student t-test, p<0.05). Parsimony analyses showed the rpoB gene evolves more than 2-fold faster than 16S rRNA. In addition, phylogeny of the rpoB gene separated each M. aeruginosa strain more clearly compared with a 16S rRNA tree. This study found that the order Chroococcales, including Microcystis, has approximately two rRNA operons and single copy of the rpoB gene in their chromosomes. These results suggest that the rpoB gene is a useful marker for the molecular phylogenetics and the detection of Microcystis.

Bacterial Community Structure and Diversity of the Zoysia japonica Soil Treated with Liquid Fertilizer Containing Amino Acids (아미노산 액비를 처리한 들잔디 토양 미생물 군집구조 및 다양성)

  • Kim Dong-Il;Kim Dong-Hun
    • Korean Journal of Microbiology
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    • v.42 no.2
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    • pp.103-110
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    • 2006
  • T-RFLP analysis and clone sequencing analysis based on bacterial 16S rDNA were conducted to assess bacterial community structure and diversity in Zoysia japonica soil treated with liquid fertilizer containing amino acids(LFcAA) after spray with herbicide. The results of T-RFLP (terminal restriction fragment length poly-morphism) analysis using restriction enzyme Hae III showed that the T-RFs of various size appeared evenly in the 32 clones of KD3 and 38 clones of KD4 respectively that had been treated with liquid fertilizer containing amino acid(LFcAA) compared to 23 clones of KD2 hat had not been treated with LFcAA. The microbial com- munity structure in KD2 appeared less diverse than those in KD3 and KD4. Analysis of partial sequences for 110 clones from KDI (control), KD2 (non-treated), KD3 (LFcAA 1X), KD4 (LFcAA 2X), respectively, revealed that most bacteria were related with uncultured bacteria in a 16S rDNA sequence similarity range of 91-99% through blast search. Otherwise, the other clones were members of proteobacteria, Acidobacteria, Act-inobacteria, Sphingobacteria and Planctomyces groups. Especially in KD4, members of Alpha Proteobacteria, Rhizobiales, Sphigomonadales, Caulobacterales, Gamma Proteobacteria, the genus Pseudomonas, Betapro-teobacteria, Nitrosomonadales and genus Nitrosospira appeared to be dominant. In addition, Acidobacteria group, Actinobacteria group, Planctomycetacia and Sphingobacteria were also shown. The microbial com-munity structure in Z. japonica soil sprayed with herbicide was affected by LFcAA.

Bacterial Community Structure and Diversity Using 16S rDNA Analysis in the Intertidal Sediment of Ganghwa Island (16S rDNA 분석을 이용한 강화도 장화리 갯벌 퇴적물 내 미생물 군집구조 및 다양성)

  • Cho Hye Youn;Lee Jung-Hyun;Hyun Jung-Ho
    • Korean Journal of Microbiology
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    • v.40 no.3
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    • pp.189-198
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    • 2004
  • T-RFLP analysis and clone sequencing analysis based on bacterial 16S rDNA were conducted to assess bacterial community structure and diversity in two layers (0-1cm, 6-7cm depth) of the sediment from Janghwari intertidal flat in Ganghwa Island. The results of T-RFLP (terminal-restriction fragment length polymorphism) analysis using restriction enzyme HhaI showed that the T-RFs of various size ($60{\pm}2$) bp-($667{\pm}2$) bp) appeared evenly at the surface sediments but two T-RFs with 60(${\pm}2$)bp and 93 (${\pm}2$)bp predominated at 6-7cm depth. Analysis of partial sequences for 172 clones revealed that 98% of the clones were not matched with the sequences of cultured bacteria strains in the GenBank (${\geq}similarity$ 98%), and approximately 86% of them were classified as different phylotypes. Most clones belonged to $\alpha$-, $\gamma$-, and $\delta$-Proteobacteria, Acidobacteria/Holophaga and green nonsulfur bacteria group. Proteobacteria group occupied the highest proportion in both layers (69% at 0-1cm depth and 46% at 6-7cm depth). $\gamma$-Proteobacteria and $\delta$-Proteobacteria that are associated with oxidation and reduction of sulfur compounds were appeared to be dominant, and comprised 21.5% and 15.7% of total clones, respectively. Overall results indicated that extremely diverse bacterial groups were inhabiting in the sediment of Ganghwa intertidal flat, and bacterial communities associated with the behaviour of sulfur seemed to playa significant role in the biogeochemical environment in this anoxic sediment.

Genetic Divergence and Phylogenetic Relationships among the Korean Fireflies, Hotaria papariensis, Luciola lateratis, and Pyrocoelia rufa(Coleoptera: Lampyridae), using Mitochondrial DNA Sequences (미토콘드리아 DNA의 염기서열을 이용한 파파리반딧불이, 애반딧불이 및 늦반딧불이 (딱정벌레목: 반딧불이과)의 유전적 분화 및 계통적 관련)

  • 김익수;이상철;배진식;진병래;김삼은;김종길;윤형주;양성렬;임수호
    • Korean journal of applied entomology
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    • v.39 no.4
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    • pp.211-226
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    • 2000
  • Genetic divergence and phylogenetic relationships among the major Korean fireflies (Hotaria papariensis, Luciola lateralis, and Pyrocoelia rufa) were studied. A portion of mitochondrial COI (403 bp) and 165 rRNA (490~504 bp) genes were sequenced, and the GenBank-registered, homologous 165 rRNA sequences of Japanese fireflies were compared (27 species of Lampyridae, one of Lycidae, and one of Rhgophthalmidae). Greatest DNA and/or amino acid sequence divergence was found when P rufa, belonging to Lampyrinae was compared with H. papariensis and L. lateralis, both belong-ing to Luciolinae, confirming the current taxonomic status of the species. In the PAUP and PHYLIP analyses with 165 rRNA data, grouping of the two geographic samples of H. papariensis with H. tsushimana validate the use of generic name, Hotaria. Nevertheless, lack of sister-group relationship of the two geographic samples of H. papariensis renders further investigation on this group . Although the Korean and Japanese L. lateralis formed a strong monophyletic group, a substantial genetic differentiation was detected between them (2.9% of 165 rRNA gene sequence divergence). Finally, the geographic samples of Korean p. rufa strongly formed a group with Japanese p. rufa, warranting the use of generic name, Pyrocoelia, but the genetic distance observed between the Cheju-Island individual and all others requires further investigation on this subject. Summarized, this study supports the current taxonomic status of the Korean fireflies in that each respectively formed a strong monophyletic group with its own species or genus.

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수계 생태계에서의 세균 군집 구조의 분자생물학적 분석

  • 이동훈;김상종
    • Korean Journal of Microbiology
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    • v.33 no.1
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    • pp.55-65
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    • 1997
  • 16S rRNA를 분석한 연구들은 자연 생태계에서 추출한 핵산을 이용하여 16rRNA 유전자의 염기서열을 분석하거나 특정 DNA probe를 이용한 hybridization 실험이 주류를 이루어 왔다. 특히 PCR 기법이 개발됨에 따라 적은 양의 시료를 대량으로 손쉽게 증폭시킬 수 있어 다양한 분야에 응용되고 있다. 세균 군집의 구조를 이해하는데 있어서 PCR 방법의 적용 대상은 주로 16S rRNA 유전자의 염기서열 해독분야이며 해양 생태계를 대상으로 많은 연구 결과가 보고되었다(11,13,21,26). 한편 자연 생태계의 개별적 미생물 분류룬들을 검출하기 위한 특정 oligonucleotide probe의 개발방법들은 미생물 군집의 유전적 다양성에 대한 정보 파악 이외에 배양이 어려운 혐기성 세균과 같은 특정 세균들의 동정에도 이용되고 있다(3,24,55). 본고에서는 세균 군집의 구조와 다양성을 연구하는데 적용 가능한 rRNA 분석방법들을 수계 생태계를 중심으로 살펴보고자 한다.

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Isolation and identification of culturable bacteria from human skin (배양가능한 피부세균의 분리 및 동정)

  • Bae, Young-Min
    • Journal of the Korean Applied Science and Technology
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    • v.37 no.6
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    • pp.1698-1705
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    • 2020
  • Bacteria were collected from the thumb surface of the twenty young adults that are 20 to 25 years old and cultured on the Luria-Bertani agar. The 16S rDNA of the cultured bacteria was amplified by polymerase chain reaction(PCR) and DNA sequence of the PCR products analyzed. Total 14 different bacterial species were identified by comparing their 16S rDNA sequence with the data in genbank. It appears that each individual has 2.5 different bacterial species in average. Staphylococcal species were the most abundant among the identified bacteria and Micrococcus luteus was the second. Staphylococcal species were isolated at similar frequency between male and female donors but Micrococcus luteus was isolated more frequently from female than male donors. The result obtained in this study might be useful in research of dermatic diseases, searching for new drugs for those diseases and development of new cosmetics.

Molecular Taxonomy based on 16S rDNA Analysis and Pathogenicity of Yersinia pseudotuberculosis Isolated from Spring Waters (약수에서 분리한 Yersinia pseudotuberculosis의 병원성과 16S rDNA 분석에 의한 분자학적 분류)

  • Lee, Young-Kee;Choi, Sung-Min;Oh, Soo-Kyung;Lee, Kang-Moon;Ryeom, Kon
    • Korean Journal of Microbiology
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    • v.37 no.1
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    • pp.9-14
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    • 2001
  • In order to investigate the pathogenicity and development of differential identification technique in the Yersinia species and other entericbacteria, we isolated 5 strains of Y.pseudotuberculosis from spring water sites in Seoul. The biochemical characteristics of isolated strains revealed that indole, VP($25^{\circ}C$, $37^{\circ}C$), $H_2S$, phenylalanine, lysine, arginine, ornithine, gas from glucose, lactose, sucrose, sorbitol, oxidase and motility($37^{\circ}C$) were all negative and urease, glucose, mannitol, salicin, catalase and motility($25^{\circ}C$) were all positive. To detect the causative agent of pseudotuberculosis(Y.pseudotuberculosis), we carried out a study using a PCR with inv primers complementary to the pathogenic region and found that all strains were positive, this revealed that strains from spring waters were pathogenic. Also 16S rDNA for total 5 strains of Y. pseudotuberculosis were amplified and a stretch of approximately 1,450 nucleotides were sequenced and analyzed. The 16S rDNA nucleotide sequence homologies among Yersinia species ranged 97.5% to 100% and between Y.pseudotuberculosis and other entericbacteria they ranged 93.0% to 95.1%. The Phylogenetic tree generated from the sequence analysis of the 16S rDNA gene showed 3 coherent clusters that could be separated into Y.pseudotuberculsis strains, some Yersinia species strains and other entericbacteria strains.

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Genetic Diversity of Rana amurensis (Amphibia: Ranidae), Based on Mitochondrial 165 rDNA Gene Sequences (미토콘드리아 16S rDNA를 이용한 아무르산개구리 (양서 강: 개구리 과)의 유전적 다양성)

  • 송재영;윤병수;오홍식;정규회
    • Korean Journal of Environmental Biology
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    • v.21 no.1
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    • pp.45-51
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    • 2003
  • Genetic diversity of local populations among geologically isolated groups of Rana amurensis was refined by sequence comparison of the mitochondrial (mt) 165 rDNA genes. Each 401 base pairs of DNA sequences, which was determined from four local populations of Rana amurensis, two local populations of R. nigromacutata, and three species of the genus Rana were used in this analysis. Despite morphological similarity of Rana amurensis, Korean populations were well distinguished from the other groups on the basis of 105 rDNA gene difference. Further analyses for additional local populations belonging to R. amurensis will be necessary to clarify the taxonomic status.