• 제목/요약/키워드: 16S rRNA gene sequencing

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메타분석을 통한 반려견 분변 박테리아 군집 조사 (A Meta-Analysis of Fecal Bacterial Diversity in Dogs)

  • 정진영;김민석
    • 한국산학기술학회논문지
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    • 제18권1호
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    • pp.141-147
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    • 2017
  • 본 연구에서는 클로닝과 생어 염기서열 분석으로 획득된 16S rRNA 유전자 염기서열을 메타분석하여 반려견 분변 박테리아를 조사하였다. 이러한 메타분석을 위해서 RDP 데이터베이스(Release 11, Update 3)에 등록되어 있는 반려견 분변 박테리아 유래 16S rRNA 유전자 염기서열 검색하여 획득하였다. RDP 데이터베이스에서 총 420개의 반려견 분변 박테리아 유래 16S rRNA 유전자 염기서열이 확인되었고, 그 중에서 42개 유전자 염기서열이 배양가능한 박테리아에서 유래한 것으로 확인되었다. 이러한 420개의 유전자 염기서열은 박테리아 분류학상의 '문'(phylum)에서 총 5개(Firmicutes, Bacteroidetes, Actinobacteria, Fusobacteria, Proteobacteria)로 분류되었다. 그 중에서 Firmicutes가 가장 우점하는 '문'이었고, 총 420개 유전자 중에서 55.2%를 차지하였다. Bacteroidetes는 32.1%로 두 번째로 우점하는 '문'이였고, 다음으로 Actinobacteria(6.4%), Fusobacteria(3.8%), Proteobacteria(2.4%)가 우점하였다. 박테리아 분류학상의 '속'(genus)에서는 Bacteroidetes의 하위 단계인 Bacteroides가 가장 우점하였고 총 420개 유전자 중에서 30.0%를 차지하였다. 반면에 Firmicutes의 하위 단계인 Clostridium XI는 두 번째로 우점하는 '속'으로 총 420개 유전자 중에서 27.4%를 차지하였다. 추정상의 '종'(species)인 Operational taxonomic units의 수는 82개로 확인되었다. 본 연구의 결과는 반려견 분변 내 미생물 다양성을 이해하는데 도움을 줄 수 있을 것이고, 향후 반려견의 건강과 웰빙에 관한 연구에 활용될 수 있을 것이다.

Occurrence of Stolbur Phytoplasma Disease in Spreading Type Petunia hybrida Cultivars in Korea

  • Chung, Bong Nam;Jeong, Myeong Il;Choi, Seung Kook;Joa, Jae Ho;Choi, Kyeong San;Choi, In Myeong
    • The Plant Pathology Journal
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    • 제29권4호
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    • pp.465-470
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    • 2013
  • In January 2012, spreading type petunia cv. Wave Pink plants showing an abnormal growth habit of sprouting unusual multiple plantlets from the lateral buds were collected from a greenhouse in Gwacheon, Gyeonggi Province, Korea. The presence of phytoplasma was investigated using PCR with the primer pairs P1/P6, and R16F1/R1 for nested-PCR. In the nested PCR, 1,096 bp PCR products were obtained, and through sequencing 12 Pet-Stol isolates were identified. Comparison of the nucleotide sequences of 16S rRNA gene of the 12 Pet-Stol isolates with other phytoplasmas belonging to aster yellows or Stolbur showed that Pet-Stol isolates were members of Stolbur. The presence of phytoplasma in petunia was also confirmed by microscopic observation of the pathogens. In this study, Stolbur phytoplasma was identified from spreading type petunia cultivars by sequence analysis of 16S rRNA gene of phytoplasma and microscopic observation of phytoplasma bodies. This is the first report of Stolbur phytoplasma in commercial Petunia hybrida cultivars.

Practical considerations for the study of the oral microbiome

  • Yu, Yeuni;Lee, Seo-young;Na, Hee Sam
    • International Journal of Oral Biology
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    • 제45권3호
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    • pp.77-83
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    • 2020
  • In the oral cavity, complex microbial community is shaped by various host and environmental factors. Extensive literature describing the oral microbiome in the context of oral health and disease is available. Advances in DNA sequencing technologies and data analysis have drastically improved the analysis of the oral microbiome. For microbiome study, bacterial 16S ribosomal RNA gene amplification and sequencing is often employed owing to the cost-effective and fast nature of the method. In this review, practical considerations for performing a microbiome study, including experimental design, molecular analysis technology, and general data analysis, will be discussed.

임상에서 분리된 희귀 비결핵 마이코박테리아 5종 (Five Rare Non-Tuberculous Mycobacteria Species Isolated from Clinical Specimens)

  • 박영길;이영주;유희경;정미영;류성원;김창기;김희진
    • Tuberculosis and Respiratory Diseases
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    • 제69권5호
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    • pp.331-336
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    • 2010
  • Background: Recently, the rate of infections with non-tuberculous mycobacteria (NTM) has been increasing in Korea. Precise identification of NTM is critical to determination of the pathogen and to target treatment of NTM patients. Methods: Sixty-eight unclassified mycobacteria isolates by rpoB PCR-RFLP assay (PRA) collected in 2008 were analyzed by National Center for Biotechnology Information (NCBI) Basic Local Alignment Search Tool (BLAST) search after sequencing of 16S rRNA, hsp65, rpoB genes. Results: Nineteen strains of 68 isolates were specified as species after sequencing analysis of 3 gene types. We found 3 M. lentifulavum, 5 M. arupense, 4 M. triviale, 4 M. parascrofulaceum, and one M. obuense. One M. tuberculosis and another M. peregrinum were mutated at the Msp I recognition site needed for rpoB PRA. The remaining 49 isolates did not coincide with identical species at the 3 kinds genes. Conclusion: Sequencing analysis of 16S rRNA, hsp65, rpoB was useful for identification of NTM unclassified by rpoB PRA.

Isolation and Identification of Noble Lactic Acid Bacteria

  • Yeo, Han-Cheol;Jang, Jin-Young;Park, Hyeong-Jun;Min, Byung-Tae;Yoo, Min
    • Quantitative Bio-Science
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    • 제37권2호
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    • pp.125-132
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    • 2018
  • In this study, noble strains of lactic acid bacteria were isolated and identified by genetic analysis of 16s rRNA. Also, pH-dependent growth curve, cholesterol assimilation ability and sugar production efficiency were measured. Lactic acid bacteria were identified to inhabit in the milks from various animals. Results of sequence analysis showed that there were differences in 16S rRNA sequence among strains and part of gene deletion was also recognized. Growth rates were varied, too, depending on the pH of the medium. Lactobacillus rhamnosus LOCK908 isolated from cow milk showed the highest growth rate and high cholesterol assimilation ability. Results of sugar fermentation tests were relatively consistent with the sequencing results. So, we propose newly isolated Lactobacillus rhamnosus LOCK908 as useful candidate for a starter of fermented beverage and probiotics. Results of this study will contribute to the isolation and identification of noble Lactic acid bacteria and to the public health.

터널에서 미학적 문제를 야기하는 진균 및 항진균 활성을 가진 탄산칼슘 형성세균의 분리와 특성 (Isolation of Fungal Deteriogens Inducing Aesthetical Problems and Antifungal Calcite Forming Bacteria from the Tunnel and Their Characteristics)

  • 박종명;박성진;김사열
    • 한국미생물·생명공학회지
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    • 제39권3호
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    • pp.287-293
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    • 2011
  • 본 연구는 터널표면의 퇴색 및 변색을 초래할 것으로 생각되는 주요 균사형진균을 결정하고, 그 진균을 방제하기 위해 부근 장소에서 분리된 세균들이 항진균 길항능을 가지는 동시에 탄산칼슘 형성을 확인하였다. 이 균주를 이용하여 모르타르에 적용했을 때 항진균 및 압축강도 증진효과를 가진 세균자원을 확보하는데 그 목적이 있다. 터널내의 오염된 지역에서 시료를 취하여 다양한 배지를 이용하여 곰팡이, 효모 및 세균을 분리하였고, 분리된 진균의 ITS-5.8S rRNA gene sequene와 세균의 16s rDNA sequence를 이용해 부분동정을 실시했다. 분리된 미생물의 터널 내 분포를 결정하였으며, 분리된 세균 5종의 탄산칼슘 형성능력을 확인하였다. 터널내 오염지역에서 가장 널리 분포하는 곰팡이인 C. sphaerospermum KNUC253 과 감수성 시험에 널리 이용되는 공시균인 A. niger KCTC6906을 대상으로 항진균 시험을 실시하였다. 터널 분리세균 5종 모두 urea-$CaCl_2$ 고체배지에서 배양했을 때 콜로니 주변부 에서 종 특이적으로 다양한 크기와 형태의 탄산칼슘을 형성함을 확인하였다. 그 중 B. aryabhatti KNUC205는 감수성 곰팡이를 대상으로 뛰어난 항진균 길항능을 보였다.

A Case of Segniliparus rugosus Pulmonary Infection in an Immunocompetent Patient with Non-cystic Fibrosis

  • Lee, Jung Yeon;Chon, Gyu Rak;Jung, Tae-Young;Sung, Heungsup;Shim, Tae Sun;Jo, Kyung-Wook
    • Tuberculosis and Respiratory Diseases
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    • 제77권5호
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    • pp.227-229
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    • 2014
  • Segniliparus species is a novel genus that is reported to be the new emerging respiratory pathogens. Here, we report a very rare case of S. rugosus pulmonary infection in an immunocompetent patient with non-cystic fibrosis. The organism was identified by 16S rRNA gene sequencing. The patient was successfully treated with antibiotics.

A report of 10 unrecorded bacterial species of Korea, isolated from agricultural soil in 2022

  • Oung Bin Lim;Ji Soo Lee;Hyosun Lee;Ki Eun Lee;In Tae Cha;Won Jae Chi;Dong-Uk Kim
    • 환경생물
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    • 제41권3호
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    • pp.215-222
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    • 2023
  • In 2022, research for native prokaryotic species in Korea reported 10 unrecorded bacterial strains affiliated to phyla Actinomycetota, Bacillota, and Pseudomonadota. The strains formed monophyletic clades with the most closely related species (with ≥98.7% sequence similarity) in the 16S rRNA gene sequencing. Among them, four species of the phylum Actinomycetota, two species of the phylum Bacillota, and four species of the phylum Pseudomonadota have not been reported in Korea, suggesting unrecorded species in Korea. Information on strains such as Gram staining reaction, colony and cell morphology, biochemical characteristics, and isolation sources were provided in the species description.

Distribution and differential expression of microRNAs in the intestinal mucosal layer of necrotic enteritis induced Fayoumi chickens

  • Rengaraj, Deivendran;Truong, Anh Duc;Ban, Jihye;Lillehoj, Hyun S.;Hong, Yeong Ho
    • Asian-Australasian Journal of Animal Sciences
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    • 제30권7호
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    • pp.1037-1047
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    • 2017
  • Objective: Despite an increasing number of investigations into the pathophysiology of necrotic enteritis (NE) disease, etiology of NE-associated diseases, and gene expression profiling of NE-affected tissues, the microRNA (miRNA) profiles of NE-affected poultry have been poorly studied. The aim of this study was to induce NE disease in the genetically disparate Fayoumi chicken lines, and to perform non-coding RNA sequencing in the intestinal mucosal layer. Methods: NE disease was induced in the Fayoumi chicken lines (M5.1 and M15.2), and non-coding RNA sequencing was performed in the intestinal mucosal layer of both NE-affected and uninfected chickens to examine the differential expression of miRNAs. Next, quantitative real-time polymerase chain reaction (real-time qPCR) was performed to further examine four miRNAs that showed the highest fold differences. Finally, bioinformatics analyses were performed to examine the four miRNAs target genes involvement in the signaling pathways, and to examine their interaction. Results: According to non-coding RNA sequencing, total 50 upregulated miRNAs and 26 downregulated miRNAs were detected in the NE-induced M5.1 chickens. While 32 upregulated miRNAs and 11 downregulated miRNAs were detected in the NE-induced M15.2 chickens. Results of real-time qPCR analysis on the four miRNAs (gga-miR-9-5p, gga-miR-20b-5p, ggamiR-196-5p, and gga-let-7d) were mostly correlated with the results of RNAseq. Overall, ggamiR-20b-5p was significantly downregulated in the NE-induced M5.1 chickens and this was associated with the upregulation of its top-ranking target gene, mitogen-activated protein kinase, kinase 2. Further bioinformatics analyses revealed that 45 of the gene targets of gga-miR-20b-5p were involved in signal transduction and immune system-related pathways, and 35 of these targets were predicted to interact with each other. Conclusion: Our study is a novel report of miRNA expression in Fayoumi chickens, and could be very useful in understanding the role of differentially expressed miRNAs in a NE disease model.

Microbiological Characteristics of Nocardia takedensis Isolated from Skin Lesion, in Korea

  • Kang, Hye-Sook;Park, Gyu-Nam;Kim, Hye-Ran;Chang, Kyung-Soo
    • 대한의생명과학회지
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    • 제23권2호
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    • pp.96-103
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    • 2017
  • Nocardia species (spp.) are opportunistic pathogen in immunocompromised hosts. The genus Nocardia contains more than 70 species. Nocardia takedensis has been recently reported as a new species of the genus Nocardia. In this study, we describes the first clinical isolate of N. takedensis from the skin lesion in Busan, Korea. For the identification of clinical isolate to the species level as N. takedensis, classical methods (colony morphology, biochemical characteristics, and antimicrobial susceptibility), molecular method (16S rRNA gene sequencing), and MS (mass spectrometry) analysis were conducted. Clinical isolates grew slowly on the culture media (5% sheep blood agar and chocolate agar) under 5% $CO_2$ condition. Especially, carotene pigmentation was detected well on the media. Using mass spectrometry, Nocardia isolate was not identified to the species level. However, molecular method based on 16S rRNA sequencing confirmed the isolate as N. takedensis correctly. N. takedensis isolate was partial positive for acid-fast bacilli on the Ziehl-Neelsen method. And it was observed to be resistance to amoxicillin/clavulanic acid and ciprofloxacin. Our results provide useful information to develop optimal identification protocol of N. takedensis in clinical diagnostic laboratories.