• Title/Summary/Keyword: 16S-rRNA

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

  • 유소영;김미광;김화숙;황호길;김평식;임성훈;오상호;민정범;국중기
    • Microbiology and Biotechnology Letters
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    • v.32 no.2
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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.

Evaluation of 16S rRNA Databases for Taxonomic Assignments Using a Mock Community

  • Park, Sang-Cheol;Won, Sungho
    • Genomics & Informatics
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    • v.16 no.4
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    • pp.24.1-24.4
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    • 2018
  • Taxonomic identification is fundamental to all microbiology studies. Particularly in metagenomics, which identifies the composition of microorganisms using thousands of sequences, its importance is even greater. Identification is inevitably affected by the choice of database. This study was conducted to evaluate the accuracy of three widely used 16S databases-Greengenes, Silva, and EzBioCloud-and to suggest basic guidelines for selecting reference databases. Using public mock community data, each database was used to assign taxonomy and to test its accuracy. We show that EzBioCloud performs well compared with other existing databases.

Identification of Nocardia seriolae by polymerase chain reaction (PCR에 의한 Nocardia seriolae의 검출)

  • Park, Myoung-Ae;Cho, Mi-Young;Kim, Myoung-Sug;Kim, Jae-Hoon;Lee, Deok-Chan
    • Journal of fish pathology
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    • v.22 no.1
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    • pp.85-90
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    • 2009
  • A method for the identification of Nocardia seriolae, the causative agent of nocardiosis in cultured fishes, using PCR was developed in the study. A PCR primer set specific to N. seriolae was designed based on 16S-23S rRNA sequence of various Nocardia species accessed in GenBank. Designed PCR primer set, Nseri-F (5'-GCA AAC TCT TCG AAC AGT CG-3') and Nseri-R (5'-GGA TAT CAG GAC TTA CCG GC-3'), amplifies the target regions of N. seriolae only, but not 4 other Nocardia species, N. asteroides, N. crassostreae, N. farcinica and N. salmonicida.

PCR-based Identification of Microorganisms in a Kefir Grain

  • Koo, Won Hoe;Seo, Min-Gook;Ahn, Jung Hoon
    • Journal of Marine Bioscience and Biotechnology
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    • v.2 no.4
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    • pp.238-244
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    • 2007
  • Nowadays many people are concerned about being healthy, and many dairy products are taken as health supplementary foods. Among dairy products, kefir, also called as Tibet mushroom, is a yogurt fermented by kefir grain, which is a mixture of lactic acid bacteria and yeasts. Although there are many empirical evidences that kefir is very influential for human body, the exact reason is not definitively discovered. Therefore, it would be useful to understand characteristics of a kefir grain and to categorize bacteria in a kefir grain. In this paper, molecular biological apparatus such as PCR, electrophoresis, PCR purification, DNA sequencing were used to identify and classify the species of lactic acid bacteria and yeast in a kefir grain. We used PCR-based identification method using 16S rRNA primer and Internal Transcribed Spacer (ITS) primer. We identified 6 different species which were selected on different medium. In addition, observation with scanning electron microscope (SEM) enabled us to grasp an external shape of the kefir grain. Although we found a limited number of microbial species, more intensive research are needed for extensive identification of microorganism species in Korean kefir grain.

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Evaluation of the Resistance of Mungbean Lines to Sprout Rot Caused by Pseudomonas species (Pseudomonas sp. 유래 녹두 부패병의 병 저항성 녹두 계통 검정)

  • Velusamy, Vijayanand;Park, Eui-Ho
    • Journal of Life Science
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    • v.22 no.7
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    • pp.987-990
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    • 2012
  • Mungbean sprout rot is one of the most serious problems of the commercial mungbean sprout industry. In this study, 70 strains of mungbean sprout rot pathogens were isolated from rotten sprouts at different time intervals. The pathogenicity of the isolated pathogens was tested. The highly pathogenic strain (YV-St-033) was identified as Pseudomonas sp. by 16S rRNA gene sequencing. In phylogenetic analysis, the YV-St-033 strain was grouped with P. mosselii, P. putita, P. fluorescens, P. entomophila, and P. lecoglossicida. The results of the 16S rRNA gene sequence analysis revealed that the YV-St-033 strain shared the highest sequence identity (more than 99%) with the P. mosselii R10 strain. The mungbean lines of Yeungnam University germplasm were screened against the YV-St-033 strain. Based on the growth rate of the sprouts after 3 days of inoculation with the pathogen, the YV148 line was highly resistant to the pathogen. The remaining lines were either partially or fully infected. The highly resistant line YV 148 is suitable for future breeding programs due to their thin sprouts and fast growing nature.

Microbial Community Analysis Isolated from Red Starfish (Certonardoa semiregularis) Gut (빨강불가사리(Certonardoa semiregularis)에서 분리된 세균의 군집구조 분석)

  • Lee, Hae-Ri;Park, So-Hyun;Kim, Dong-Hwi;Moon, Kyung-Mi;Heo, Moon-Soo
    • Journal of Life Science
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    • v.28 no.8
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    • pp.955-961
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    • 2018
  • Although much research has focused on various bioactive substances in starfish, research on microorganisms isolated from starfish is lacking as compared with other natural products. In this study, we investigated bacterial communities in the gut of red starfish (Certonardoa semiregularis) in Jeju Island. In total, 103 bacterial strains were isolated using marine agar and R2A medium. The isolated strains were analyzed and identified using the 16S rRNA gene sequence. Based on an analysis of this gene sequence, the 103 isolated bacteria were classified into four major groups: Proteobacteria (93%: Alpha-proteobacteria, 24.8%; Beta-proteobacteria, 4%; Gammaproteobacteria, 65%) Bacteroidetes (4%), Actinobacteria (2%), and Firmicutes (1%). In addition, the isolates were divided into seven classes (Actinobacteria, Flavobacteria, Bacilli, Sphingobacteria, Alphaproteobacteria, Betaproteobacteria, and Gammaproteobacteria), 15 orders, 19 families, and 24 genera. A phylogenetic analysis revealed two strains, Lysobacter sp. and Pedobacter sp., with similarity of 97.55% and 97.58%, respectively. As the similarity in the 16S rRNA gene sequence was 98% or less compared to previously identified bacteria, the two strains may possibly be classified as a new genus or species. We suggest that additional studies, including biochemical and morphological tests, should be performed to identify the new candidate strains.

Diversity of Butyrivibrio Group Bacteria in the Rumen of Goats and Its Response to the Supplementation of Garlic Oil

  • Zhu, Zhi;Hang, Suqin;Mao, Shengyong;Zhu, Weiyun
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.2
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    • pp.179-186
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    • 2014
  • This study aimed to investigate the diversity of the Butyrivibrio group bacteria in goat rumen and its response to garlic oil (GO) supplementation as revealed by molecular analysis of cloned 16S rRNA genes. Six wethers fitted with ruminal fistulas were assigned to two groups for a cross-over design with 28-d experimental period and 14-d interval. Goats were fed a basal diet without (control) or with GO ruminal infusion (0.8 g/d). Ruminal contents were used for DNA extraction collected before morning feeding on d 28. A total bacterial clone library was firstly constructed by nearly full-length 16S rRNA gene cloned sequences using universal primers. The resulting plasmids selected by Butyrivibrio-specific primers were used to construct a Butyrivibrio group-specific bacterial clone library. Butyrivibrio group represented 12.98% and 10.95% of total bacteria in control and GO group, respectively. In libraries, clones were classified to the genus Pseudobutyrivibrio, Butyrivibrio and others within the family Lachnospiraceae. Additionally, some specific clones were observed in GO group, being classified to the genus Ruminococcus and others within the family Ruminococcaceae. Based on the criterion that the similarity was 97% or greater with database sequences, there were 29.73% and 18.42% of clones identified as known isolates (i.e. B. proteoclasticus and Ps. ruminis) in control and GO groups, respectively. Further clones identified as B. fibrisolvens (5.41%) and R. flavefaciens (7.89%) were specifically found in control and GO groups, respectively. The majority of clones resembled Ps. ruminis (98% to 99% similarity), except for Lachnospiraceae bacteria (87% to 92% similarity) in the two libraries. The two clone libraries also appeared different in Shannon diversity index (control 2.47 and GO group 2.91). Our results indicated that the Butyrivibrio group bacteria had a complex community with considerable unknown species in the goat rumen.

Community Analysis of Nitrite-Oxidizing Bacteria in Lab-Scale Wastewater Treatment System (폐수처리장치에서의 아질산염 산화 세균 군집 분석)

  • Jeong, Soon-Jae;Lee, Sang-Ill;Lee, Dong-Hun
    • Korean Journal of Microbiology
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    • v.44 no.1
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    • pp.29-36
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    • 2008
  • Nitrogen is one of the major pollutants that should be removed by wastewater treatment systems. Biological nitrogen removal (BNR) is a key technology in advanced wastewater treatment systems operated by bacterial populations. Nitrification is the first step of microbiological processes in BNR system. Ammonia is oxidized to nitrite by ammonia-oxidizing bacteria (AOB) and then nitrite is subsequently oxidized to nitrate by nitrite-oxidizing bacteria (NOB). The diversity of NOB in nitrification reactors of 3 BNR systems, Edited biological aerated filter system, Nutrient removal laboratory system, and the Rumination type sequencing batch reactor system, was investigated by terminal restriction fragment length polymorphism (T-RFLP) analysis of 16S rRNA genes. Cluster analysis of T-RF profiles showed that communities of Nitrobacter group in each system were different depending upon the process of systems. However, the clusters of Nitrospira group were divided by the habitat of aqueous and solid samples.

Differentially expressed mRNAs and their upstream miR-491-5p in patients with coronary atherosclerosis as well as the function of miR-491-5p in vascular smooth muscle cells

  • Ding, Hui;Pan, Quanhua;Qian, Long;Hu, Chuanxian
    • The Korean Journal of Physiology and Pharmacology
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    • v.26 no.3
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    • pp.183-193
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    • 2022
  • MicroRNAs (miRNAs) regulate gene expression and are biomarkers for coronary atherosclerosis (AS). A novel miRNA-mRNA regulation network of coronary AS still needs to be disclosed. The aim of this study was to analyze potential mRNAs in coronary AS patients and the role of their upstream miR-491-5p in vascular smooth muscle cells (VSMCs). We first confirmed top ten mRNAs according to the analysis from Gene Expression Omnibus database (GSE132651) and examined the expression levels of them in the plaques and serum from AS patients. Five mRNAs (UBE2G2, SLC16A3, POLR2C, PNO1, and AMDHD2) presented significantly abnormal expression in both plaques and serum from AS patients, compared with that in the control groups. Subsequently, they were predicted to be targeted by 11 miRNAs by bioinformatics analysis. Among all the potential upstream miRNAs, only miR-491-5p was abnormally expressed in the plaques and serum from AS patients. Notably, miR-491-5p overexpression inhibited viability and migration, and significantly increased the expression of contractile markers (α-SMA, calponin, SM22α, and smoothelin) in VSMCs. While silencing miR-491-5p promoted viability and migration, and significantly suppressed the expression of α-SMA, calponin, SM22α, and smoothelin. Overall, miR-491-5p targeted UBE2G2, SLC16A3, and PNO1 and regulated the dysfunctions in VSMCs.

Halobacillus blutaparonensis sp. nov., a Moderately Halophilic Bacterium Isolated from Blutaparon portulacoides Roots in Brazil

  • Barbosa Deyvison Clacino;Bae Jin-Woo;Weid Irene Von Der;Vaisman Natalie;Nam Young-Do;Chang Ho-Won;Park Yong-Ha;Seldin Lucy
    • Journal of Microbiology and Biotechnology
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    • v.16 no.12
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    • pp.1862-1867
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    • 2006
  • A moderately halophilic, Gram-positive, spore-forming bacterium was isolated from the roots of Blutaparon portulacoides, a plant found in sandy soil parallel to the beach line in Restinga de Jurubatiba, Rio de Janeiro, Brazil. The strain, designated $M9^T$, was motile and strictly aerobic with rod-shaped cells. It grew in the absence of NaCl and up to 20% NaCl, and was able to hydrolyze casein and starch. Strain $M9^T$ had a cell-wall peptidoglycan based on L-Orn-D-Asp, the predominant menaquinone present was menaquinone-7 (MK-7), diaminopimelic acid was not found, and anteiso-$C_{15:0}$ and iso-$C_{15:0}$ were the major fatty acids. A phylogenetic analysis based on 16S rRNA gene sequences showed that strain $M9^T$ belonged to the genus Halobacillus and exhibited 16S rRNA gene similarity levels of 97.8-99.4% with the type strains of the other nine Halobacillus species. The DNA-DNA relatedness of strain $M9^T$ with H. trueperi, the closest relative as regards 16S rRNA gene similarity, and H. locisalis was 21% and 18%, respectively. Therefore, on the basis of phenotypic, genotypic, and phylogenetic data, strain $M9^T$ (=ATCC BAA-$1217^T$, =CIP $108771^T$, =KCTC $3980^T$) should be placed in the genus Halobacillus as a member of a novel species, for which the name Halobacillus blutaparonensis sp. nov. is proposed.