• 제목/요약/키워드: rDNA sequencing

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Analysis of Bacterial Community Structure in Bulk Soil, Rhizosphere Soil, and Root Samples of Hot Pepper Plants Using FAME and 16S rDNA Clone Libraries

  • Kim, Jong-Shik;Kwon, Soon-Wo;Jordan, Fiona;Ryu, Jin-Chang
    • Journal of Microbiology and Biotechnology
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    • 제13권2호
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    • pp.236-242
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    • 2003
  • A culture-independent and -dependent survey of the bacterial community structure in the rhizosphere and soil samples from hot pepper plants was conducted using 16S rDNA clone library and FAME analyses. Out of the 78 clones sequenced, 56% belonged to Proteobacteria, 4% to high G+C Gram- positive group, 3% to Cytophyga-Flexibacter-Bacreroides, and 32% could not be grouped with any known taxonomic division. Among the 127 FAME isolates identified, 66% belonged to low G+C Gram-positive bacteria (Baciilus spp.) and 26% to high G+C Gram-positive bacteria. In a cluster analysis, the results for both methods were found to be strikingly dissimilar. The current study is the first comparative study of FAME and 165 rDNA clonal analyses performed on the same set of soil, rhizosphere soil, and root samples.

Molecular Phylogeny of Poecilostome Copepods Based on the 18S rDNA Sequences

  • Kim, Jihee;Kim, Won
    • Animal cells and systems
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    • 제4권3호
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    • pp.257-261
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    • 2000
  • To elucidate phylogenetic relationships among poecilostome families 18S rDNA sequence data were generated for seven poecilostome and one cyclopoid copopods by PCR cloning and sequencing techmiques. Phylogenetic trees were constructed by maximum parsimony, neighbor joining, and maximum likelihood methods using cyclopoid sequence as an outgroup. The results from three different analyses showed that the seven poecilostome families were eiridel into two groups: Clausidiidae-Myicolidae-Synaptiphillidae-bomolochidae and Lichomologidae-Chondracanthidae-Ergasilidae. The molecular phylogenies were consistent with those from the morphological characters. Therefore, these analyses porvide further evidence for the utility of 18S rDNA sequences in addressing phylogenetic relationships among poecilostome families.

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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.

Production of Bacterial Cellulose by Gluconacetobacter hansenii PJK Isolated from Rotten Apple

  • Park, Joong-Kon;Park, Youn-Hee;Jung, Jae-Yong
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제8권2호
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    • pp.83-88
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    • 2003
  • A cellulose-producing strain isolated from rotten apples was identified as Gluconacetobacter hansenii based on its physiological properties and the 16S rDNA complete sequencing method, and specifically named Gluconacetobacter hansenii PJK. The amount of bacterial cellulose (BC) produced by G. hansenii PJK in a shaking incubator was 1.5 times higher than that produced in a static culture. The addition of ethanol to the medium during cultivation enhanced the productivity of bacterial cellulose, plus the supplementation of 1% ethanol into the culture medium made the produced BC aggregate into a big lump and thus protected the bacterial-cellulose-producing G. hansenii PJK cells in the shear stress field from being converted into non-cellulose-producing (Cel) mutants. Cells subcultured three times in a medium containing ethanol retained their ability to produce BC without any loss in the production yield.

Identification of Aspergillus nidulans from cooked eggs produced by permitted factory

  • Park, Hye-Won;Kim, Kook-Joo;Youk, Ji-Hea;Woo, Jong-Tae
    • 한국동물위생학회지
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    • 제30권2호
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    • pp.283-286
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    • 2007
  • Fungus generally doesn't produce toxic or harmful substances so it has low chances to cause food poisoning. However it leads to change appearance, odor and characteristics of the contaminated foods and result in sanitary risk problems. Therefore the contamination of fungi should be prevented since they are not proper for human consumption. Green fungi with white outline raised from the air cell of cooked eggs which were collected by Gyeongi Livestock Veterinary Service in August, 2006. The results came out after the cultivation using Sabouraud's Dextrose Agar(SDA). The conidium appeared white and monospore, the shape of colony was round and oval. Conidiophore was brown and granulated and wrinkles and formed. It was confirmed as Aspergillus nidulans based on the dying using Lactophenol cotton blue, the observation of septum and vesicle from the grown spores, and rDNA sequencing.

돼지 mtDNA D-loop 지역의 Large White 특이 중복현상 탐지 (Detection of a Large White-Specific Duplication in D-loop Region of the Porcine MtDNA)

  • 김재환;한상현;이성수;고문석;이정규;전진태;조인철
    • 생명과학회지
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    • 제19권4호
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    • pp.467-471
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    • 2009
  • 돼지 6품종(Landrace, Duroc, Large White, 한국재래돼지, Berkshire, Hampshire)을 대상으로 기존에 보고된 서열을 바탕으로 제작한 primer를 이용하여 mtDNA D-loop 전체영역을 증폭하였다. 증폭된 PCR product를 cloning 및 DNA sequencing, 다중염기서열비교를 통하여 분석한 결과, mtDNA에서 heteroplasmy가 나타나는 D-loop 내 tandem repeat region 이후에 11-bp 중복이 존재하는 것을 확인하였다. 이런 중복현상은 일본재래돼지와 Duroc에서 보고되었지만, 이를 이용한 돼지 품종별 중복현상의 빈도 및 분포에 관한 연구는 이루어져있지 않다. 품종별 11-bp 중복현상을 분석하기 위해서 6품종을 대상으로 중복지역을 포함한 약 150 bp 절편을 증폭하였으며, PAGE 방법을 통하여 분석하였다. 그 결과 본 연구에서 사용한 품종들 중 모든 Large White에서 중복현상이 발생하는 것을 확인하였으며, Duroc인 경우 11.2% (9/80)에서 중복현상이 확인되었다. 반면에 Landrace, 한국재래돼지, Berkshire 및 Hampshire에서는 전혀 발견되지 않았다. 이런 결과로서, 11-bp 중복현상의 분석은 현재 구별이 불가능한 Landrace와 Large White를 구별할 수 있는 유용한 DNA marker로서 사용이 가능할 것이다.

한국인 우식아동으로부터 분리한 Streptococcus mutans의 내산성 단백질의 발현 (Acid Stress-Induced Proteins of the Streptococcus mutans Isolated from Korean Children with Caries)

  • 강경희
    • 한국산학기술학회:학술대회논문집
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    • 한국산학기술학회 2009년도 춘계학술발표논문집
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    • pp.3-4
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    • 2009
  • 한국인 아동의 우식치아로부터 S. mutans를 분리하고, acid stress하에서 분리한 S. mutans의 내산성 능력과 관련된 단백질을 규명하고자 하였다. S. mutans의 16S rDNA 유전자 핵산염기서열을 바탕으로 설계된 프라이머 쌍을 이용하여 중합효소연쇄반응을 실시하고 16S rDNA sequencing을 실시한 결과, 한국인 아동으로부터 분리 된 S. mutans K7은 기존에 보고된 표준균주와 99.9% 일치하는 결과를 나타내 S. mutans로 판명되었다. 또한 2D gel electrophoresis 를 수행한 결과, acid stress에 관여하는 것으로 추정되는 단백질들을 확인 할 수 있었다.

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Development of a Novel Long-Range 16S rRNA Universal Primer Set for Metagenomic Analysis of Gastrointestinal Microbiota in Newborn Infants

  • Ku, Hye-Jin;Lee, Ju-Hoon
    • Journal of Microbiology and Biotechnology
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    • 제24권6호
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    • pp.812-822
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    • 2014
  • Metagenomic analysis of the human intestinal microbiota has extended our understanding of the role of these bacteria in improving human intestinal health; however, a number of reports have shown that current total fecal DNA extraction methods and 16S rRNA universal primer sets could affect the species coverage and resolution of these analyses. Here, we improved the extraction method for total DNA from human fecal samples by optimization of the lysis buffer, boiling time (10 min), and bead-beating time (0 min). In addition, we developed a new long-range 16S rRNA universal PCR primer set targeting the V6 to V9 regions with a 580 bp DNA product length. This new 16S rRNA primer set was evaluated by comparison with two previously developed 16S rRNA universal primer sets and showed high species coverage and resolution. The optimized total fecal DNA extraction method and newly designed long-range 16S rRNA universal primer set will be useful for the highly accurate metagenomic analysis of adult and infant intestinal microbiota with minimization of any bias.

16S rDNA 클론 Libraries를 이용한 치근단 농양 병소의 세균 동정 (Identification of Bacteria from Periapical Abscess Using 16S rDNA Clone Libraries.)

  • 유소영;김미광;김화숙;황호길;김평식;임성훈;오상호;민정범;국중기
    • 한국미생물·생명공학회지
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    • 제32권2호
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    • pp.195-198
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    • 2004
  • Molec-ular analysis was performed on the microflora found In the necrotic pulpal tissue collected from 5 infected root canals that were diagnosed as a periapical abscess. 16S rRNA coding gene (rDNA) library construction and sequencing were performed in order to identify the microflora, The 16S rDNA sequences from 278 clones were identified by a comparison with the database sequence in GenBank. Three phylum and 31 species, which were related to the oral microflora, were identified from the 3 samples (No. 87, 105, and 115). Dialister invisus (5.6%), Peptostreptococcus micron (18.3%), and Veillonella sp. (3.3%) were the organism present in all tee samples. Lac-tobacillusfementum (2.8%),Eubacterumsp./E. infirmum (6.7%), Shuttleworthiasatelles (3.9%), Psudorarnihacfer alactoiyticus (13.3%), Bulleidia moorei (2.8%), and Prevotella denticola (1.1%) were found in two samples. Two phylum and low species of environmental microflora were identified from 2 samples (No.95 and 101). The reason for this might be contamination of the samples with dental water. These results showed that molecular analysis could reveal more diverse microflora that are associated with endodontic infections than that revealed by conventional cultural methods. In addition, these results may of for the basic data to epidemiological studies related with endodontic infection.