• Title/Summary/Keyword: rRNA sequencing

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Septicemia Caused by Leuconostoc lactis with Intrinsic Resistance to Vancomycin in a Patient with Biliary Stent

  • Shin, Kyeong Seob;Han, Kyudong;Hong, Seung Bok
    • Biomedical Science Letters
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    • v.19 no.3
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    • pp.280-283
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    • 2013
  • Leuconostoc spp. is intrinsically resistant against vancomycin and rarely causes the infection in immunocompromised patients. In this report, we describe a fatal case of Leuconostoc lactis bacteremia in a patient with biliary tract stent insertion to resolve the biliary tract obstruction by multiple pseudocysts in the pancreatic head region. Leuconostic lactis isolated from the blood of the patients was confirmed by 16S rRNA sequencing and this isolate was susceptible against most antibiotics, including levofloxacin, penicillin, erythromycin and cefotaxime except vancomycin. The septic shock and multi-organ failure was abruptly progressed due to delayed use of adequate antibiotic. Using vancomycin as the empirical antibiotics in a bacteremic patient by Gram positive cocci, the treatment failures by the isolates with intrinsic resistance against vancomycin have to be considered. In addition, the prompt and accurate identification of Leuconostoc spp. are very important to select the adequate antibiotics.

Isolation and Identification of Lactic Acid Bacteria Isolated from a Traditional Jeotgal Product in Korea

  • Cho, Gyu-Sung;Do, Hyung-Ki
    • Ocean Science Journal
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    • v.41 no.2
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    • pp.113-119
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    • 2006
  • Seventeen lactic acid bacterial strains (LAB) were isolated using MRS agar medium from Jeotgal, a Korean fermented food, purchased at the Jukdo market of Pohang. To identify the strains isolated, they were tested by examining their cell morphologies, gram-staining, catalase activity, arginine hydrolase activity, D-L lactate form and carbohydrate fermentation. According to the phenotypic characteristics, three strains were tentatively identified as Lactobacillus spp., ten were Enterococcus spp. (or Streptococcus spp., or Pediococcus spp.) and the rest were Leuconostoc spp. (or Weissella spp.). Five strains among 17 were chosen by preliminary bacteriocin activity test. Four bacterial strains which inhibited both indicator microorganisms were identified by 16S rRNA sequencing. The results are as follows; Leuconostoc mesenteroides (HK 4), Leuconostoc mesenteroides (HK 5), Leuconostoc mesenteroides(HK 11), Streptococcus salivarius(HK 8). In order to check LAB which are showing a high survival rate in gut, we investigated three strains inhibiting both indicator microorganisms in artificial gastric acid and bile juice -all except HK8. The three strains mentioned above grew in extreme low acid conditions.

hnRNPK-regulated PTOV1-AS1 modulates heme oxygenase-1 expression via miR-1207-5p

  • Shin, Chang Hoon;Ryu, Seongho;Kim, Hyeon Ho
    • BMB Reports
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    • v.50 no.4
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    • pp.220-225
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    • 2017
  • Antisense transcripts were initially identified as transcriptional noise, but have since been reported to play an important role in the quality control of miRNA functions. In this report, we tested the hypothesis that heterogeneous nuclear ribonucleoprotein K (hnRNPK) regulates miRNA function via competitive endogenous RNAs, such as pseudogenes, long non-coding RNAs, and antisense transcripts. Based on analyses of RNA sequencing data, the knockdown of hnRNPK decreased the antisense PTOV1-AS1 transcript which harbors five binding sites for miR-1207-5p. We identified heme oxygenase-1 (HO-1) mRNA as a novel target of miR-1207-5p by western blotting and Ago2 immunoprecipitation. The knockdown of hnRNPK or PTOV1-AS1 suppressed HO-1 expression by increasing the enrichment of HO-1 mRNA in miR-1207-5p-mediated miRISC. Downregulation of HO-1 by a miR-1207-5p mimic or knockdown of hnRNPK and PTOV1-AS1 inhibited the proliferation and clonogenic ability of HeLa cells. Taken together, our results demonstrate that hnRNPK-regulated PTOV1-AS1 modulates HO-1 expression via miR-1207-5p.

IVAG: An Integrative Visualization Application for Various Types of Genomic Data Based on R-Shiny and the Docker Platform

  • Lee, Tae-Rim;Ahn, Jin Mo;Kim, Gyuhee;Kim, Sangsoo
    • Genomics & Informatics
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    • v.15 no.4
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    • pp.178-182
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    • 2017
  • Next-generation sequencing (NGS) technology has become a trend in the genomics research area. There are many software programs and automated pipelines to analyze NGS data, which can ease the pain for traditional scientists who are not familiar with computer programming. However, downstream analyses, such as finding differentially expressed genes or visualizing linkage disequilibrium maps and genome-wide association study (GWAS) data, still remain a challenge. Here, we introduce a dockerized web application written in R using the Shiny platform to visualize pre-analyzed RNA sequencing and GWAS data. In addition, we have integrated a genome browser based on the JBrowse platform and an automated intermediate parsing process required for custom track construction, so that users can easily build and navigate their personal genome tracks with in-house datasets. This application will help scientists perform series of downstream analyses and obtain a more integrative understanding about various types of genomic data by interactively visualizing them with customizable options.

Detection and Potential Abundances of Anammox Bacteria in the Paddy Soil

  • Khanal, Anamika;Lee, Seul;Lee, Ji-Hoon
    • Korean Journal of Environmental Agriculture
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    • v.39 no.1
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    • pp.26-35
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    • 2020
  • BACKGROUND: Microbes that govern a unique biochemical process of oxidizing ammonia into dinitrogen gas, such as anaerobic ammonium oxidation (anammox) have been reported to play a pivotal role in agricultural soils and in oceanic environments. However, limited information for anammox bacterial abundance and distribution in the terrestrial habitats has been known. METHODS AND RESULTS: Phylogenetic and next-generation sequencing analyses of bacterial 16S rRNA gene were performed to examine potential anammox bacteria in paddy soils. Through clone libraries constructed by using the anammox bacteria-specific primers, some clones showed sequence similarities with Planctomycetes (87% to 99%) and anammox bacteria (94% to 95%). Microbial community analysis for the paddy soils by using Illumina Miseq sequencing of 16S rRNA gene at phylum level was dominated by unclassified Bacteria at 33.2 ± 7.6%, followed by Chloroflexi at 20.4 ± 2.0% and Acidobacteria at 17.0 ± 6.5%. Planctomycetes that anammox bacteria are belonged to was 1.5% (± 0.3) on average from the two paddy soils. CONCLUSION: We suggest evidence of anammox bacteria in the paddy soil. In addition to the relatively well-known microbial processes for nitrogen-cycle, anammox can be a potential contributor on the cycle in terrestrial environments such as paddy soils.

Molecular identification of coagulase-negative staphylococci by rpoB sequence typing (rpoB 염기서열 분석을 이용한 응고효소 음성 포도알세균 분자 동정)

  • Seong, Won-Jin;Kim, Danil;Kim, Eun-Kyung;Ko, Dae-Sung;Ro, Younghye;Kim, Jae-Hong;Kwon, Hyuk-Joon
    • Korean Journal of Veterinary Research
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    • v.58 no.1
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    • pp.51-55
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    • 2018
  • Bovine mastitis (BM) has resulted in enormous economic loss in the dairy industry and coagulase-negative staphylococci (CNS) have caused subclinical BM. Although VITEK 2 GP ID card (VITEK 2) has been used for CNS identification, the probability of identification varies. The rpoB sequence typing (RSTing) method has been used for molecular diagnosis and epidemiology of bacterial infections. In this study, we undertook RSTing of CNS and compared the results with those of VITEK2 and 16S rRNA gene sequencing. As compared VITEK2, the molecular-based methods were more reliable for species identification; moreover, RSTing provided more molecular epidemiological information than that from 16S rRNA gene sequencing.

Metagenomic SMRT Sequencing-Based Exploration of Novel Lignocellulose-Degrading Capability in Wood Detritus from Torreya nucifera in Bija Forest on Jeju Island

  • Oh, Han Na;Lee, Tae Kwon;Park, Jae Wan;No, Jee Hyun;Kim, Dockyu;Sul, Woo Jun
    • Journal of Microbiology and Biotechnology
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    • v.27 no.9
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    • pp.1670-1680
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    • 2017
  • Lignocellulose, composed mostly of cellulose, hemicellulose, and lignin generated through secondary growth of woody plant, is considered as promising resources for biofuel. In order to use lignocellulose as a biofuel, biodegradation besides high-cost chemical treatments were applied, but knowledge on the decomposition of lignocellulose occurring in a natural environment is insufficient. We analyzed the 16S rRNA gene and metagenome to understand how the lignocellulose is decomposed naturally in decayed Torreya nucifera (L) of Bija forest (Bijarim) in Gotjawal, an ecologically distinct environment. A total of 464,360 reads were obtained from 16S rRNA gene sequencing, representing diverse phyla; Proteobacteria (51%), Bacteroidetes (11%) and Actinobacteria (10%). The metagenome analysis using single molecules real-time sequencing revealed that the assembled contigs determined originated from Proteobacteria (58%) and Actinobacteria (10.3%). Carbohydrate Active enZYmes (CAZy)- and Protein families (Pfam)-based analysis showed that Proteobacteria was involved in degrading whole lignocellulose, and Actinobacteria played a role only in a part of hemicellulose degradation. Combining these results, it suggested that Proteobacteria and Actinobacteria had selective biodegradation potential for different lignocellulose substrates. Thus, it is considered that understanding of the systemic microbial degradation pathways may be a useful strategy for recycle of lignocellulosic biomass, and the microbial enzymes in Bija forest can be useful natural resources in industrial processes.

Identification of Cryptosporidium in Environmental Sample using Nested PCR-RFLP and DNA Sequencing (Nested PCR-RFLP 및 DNA Sequencing을 이용한 환경시료에서의 크립토스포리디움 동정)

  • Park, Sangjung;Jeong, Hyanghee
    • Journal of Korean Society on Water Environment
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    • v.24 no.6
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    • pp.817-822
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    • 2008
  • In order to identify various Cryptosporidium species in environment, nested PCR-RFLP and DNA sequencing method were used. The sensitivity of nested PCR-RFLP based on 18s rRNA gene was shown to 1 oocyst. Therefore, we applied nested PCR-RFLP method to environmental samples. As a result, only 4 samples out of 8 samples confirmed as Cryptosporidium parvum by standard method of Cryptosporidium were identified as Cryptosporidium parvum by nested PCR-RFLP and DNA sequencing method. The rest of 4 samples among 8 samples were identified as Cryptosporidium muris, Cryptosporidium bailey. Therefore, in addition to standard method of Cryptosporidium, supplementary verification through nested PCR-RFLP and DNA sequencing should be needed to give more accurate information about risk of Cryptosporidium.

Genetic characterization and phylogenetic analysis of Clostridium chauvoei isolated from Hanwoo in Jeonbuk (전북지역 한우에서 분리한 기종저 균의 유전학적 특성 규명)

  • Kim, Chul-Min;Jeong, Jae-Myong;Choi, Ki-Young
    • Korean Journal of Veterinary Service
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    • v.37 no.3
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    • pp.157-164
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    • 2014
  • Clostridium chauvoei is the etiologic agent of blackleg, a high mortality rated disease infection mainly cattle. In the present study, the partial sequences of 16S rRNA and flagellin gene of C. chauvoei isolated in Jeonbuk, Korea were determined and compared with those of reference strain. Oligonucleotide primers were designed to amplify a 811 bp fragment of 16S rRNA gene and 1229 bp fragment of flagellin gene. Sequencing analysis of 16S rRNA gene showed high homology to the reference strains ranging 82.3% to 100%, while flagellin gene were different from published foreign clostridia, showing 98.7% to 72.0% nucleotide sequence homology. Phylogenetic analysis based on 16S rRNA gene revealed the close phylogenetic relationship of C. chauvoei and C. septicum in cluster I, which includes C. carnis, C. tertium, C. quinii, C. celatum, C. perfringens, C. absonum, C. botulinum B. Phylogentic analysis also revealed that flagellin gene formed a single cluster with C. chauvoei, C. septicum, C. novyi A, C. novyi B, C. tyrobutylicum, C. acetobutylicum. The genetic informations obtained from this study could be useful for the molecular study of C. chauvoei.

Isolation of Major Microflora Bacillus coagulans from Rice Bran (쌀겨발효로부터 분리된 우점종 미생물 Bacillus coagulans의 분리)

  • Lee, Sang-Han;Park, Po
    • Food Science and Preservation
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    • v.17 no.1
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    • pp.165-168
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    • 2010
  • It is known that temperature during solid fermentation using rice bran is increased upto 60-70 degree. To investigate the major microflora regarding temperature maintenance of rice bran bathing, we first isolated predominant microbes using various media by a limiting dilution method. The RNA of isolated strains were purified and sequenced. The rRNA sequencing revealed that the selected strains were similar to Bacillus coagulans according to their taxonomical relationships. Together, these results indicate that Bacillus coagulans is a major kind of microflora during solid fermentation using rice bran.