• Title/Summary/Keyword: 16S rRNA Gene

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Analysis of Microbial Diversity in Makgeolli Fermentation Using PCR-DGGE (PCR-DGGE를 이용한 막걸리발효에서 미생물 다양성 분석)

  • Kwon, Seung-Jik;Ahn, Tae-Young;Sohn, Jae-Hak
    • Journal of Life Science
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    • v.22 no.2
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    • pp.232-238
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    • 2012
  • Kumjungsansung-Makgeolli$^{(R)}$ is a traditional Korean rice wine that is fermented from traditional nuruk and rice. In this study, we performed the PCR-denaturing gradient gel electrophoresis (DGGE) analysis targeting the 16S and 28S rRNA genes to characterize bacterial and fungal diversity during Makgeolli fermentation. The predominant bacteria in the PCR-DGGE profile during Makgeolli fermentation were Lactobacillus spp. (Lactobacillus curvatus, L. kisonensis, L. plantarum, L. sakei, and L. gasseri), Pediococcus spp. (P. acidilactici, P. parvulus, P. agglomerans, and P. pentosaceus), Pantoea spp. (P. agglomerans and P. ananatis), and Citrobacter freundii; these were identified on the base of analysis of 16S rRNA gene sequences. The dominant bacterium during Makgeolli fermentation was L. curvatus. The predominant fungi in PCR-DGGE profile during Makgeolli fermentation were Pichia kudriavzevii, Saccharomyces cerevisiae, Asidia idahoensis, Kluyveromyces marxianus, Saccharomycopsis fibuligera, and Torulaspora delbrueckii, and these were identified on the basis of analysis of 28S rRNA gene sequences. The dominant fungal species during Makgeolli fermentation changed from P. kudriavzevii at 0-2 days incubation to S. cerevisiae at 3-6 days incubation. This study suggests that PCR-DGGE analysis could be a suitable tool for the understanding of microbial diversity and structure during Makgeolli fermentation.

Isolation of Cyanobacteria Producing Microcystin from Lakes (담수 생태계에서 Microcystin을 생산하는 남조세균의 분리)

  • Lee, Hee-Seon;Oh, Kyoung-Hee;Cho, Young-Cheol
    • Korean Journal of Microbiology
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    • v.44 no.3
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    • pp.251-257
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    • 2008
  • Four potential microcystin-producing cyanobacteria were isolated from large reservoirs which act as sources of drinking water supply in Korea. Strain DC-2, YD-l, and YD-6 were closely related to Microcystis aeruginosa based on the analysis of l6S rRNA gene and mcyA gene sequences. mcyA gene sequence of YDS2-3, isolated from Yongdam Reservoir, was closed to that of M. aeruginosa, whereas l6S rRNA gene sequence was not related to the known sequences of microcystin-producing cyanobacteria indicating this strain can be a novel cyanobacterium belonging to the genus Microcystis. When mcyA gene sequences of isolated cyanobacteria were compared with the mcyA gene sequence library of two reservoirs, the sequence of DC-2 matched with the dominant ones.

Development and Validation of Quick and Accurate Cephalopods Grouping System in Fishery Products by Real-time Quantitative PCR Based on Mitochondrial DNA (두족류의 진위 판별을 위한 Real-time Quantitative PCR 검사법 개발 및 검증)

  • Chung, In Young;Seo, Yong Bae;Yang, Ji Young;Kwon, Ki sung;Kim, Gun Do
    • Journal of Food Hygiene and Safety
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    • v.33 no.4
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    • pp.280-288
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    • 2018
  • In this study, an approach for the analysis of the five cephalopod species (octopus, long-arm octopus, squid, wet-foot octopus, beka squid) consumed in the Republic of Korea is developed. The samples were collected from the Southeast Asian countries Thailand, Indonesia, Vietnam, and China. The SYBR-green-based real-time qPCR method, based on the mitochondrial DNA genome of the five cephalopods was developed and validated. The intergroup variations in the mitochondrial DNA are evident in the bioinformatic analysis of the mitochondrial genomic DNA sequences of the five groups. Some of the highly-conserved and slightly-variated regions are identified in the mitochondrial cytochrome-c-oxidase subunit I (COI) gene, 16s ribosomal RNA (16s rRNA) gene, and 12s ribosomal RNA (12s rRNA) gene of these groups. To specify each five cephalopod groups, specific primer sets were designed from the COI, 16s rRNA and 12s rRNA regions. The specific primer sets amplified the DNA using the SYBR-green-based real-time PCR system and 11 commercially secured animal tissues: Octopus vulgaris, Octopus minor, Todarodes pacificus, Dosidicus gigas, Sepia esculenta, Amphioctopus fangsiao, Amphioctopus aegina, Amphioctopus marginatus, Loliolus beka, Loligo edulis, and Loligo chinensis. The results confirmed by a conveient way to calculate relative amplification levels between different samples in that it directly uses the threshold cycles (Ct)-value range generated by the qPCR system from these samples. This genomic DNA-based molecular technique provides a quick, accurate, and reliable method for the taxonomic classification of the animal tissues using the real-time qPCR.

Bacterial Diversity at Different Depths in Lead-Zinc Mine Tailings as Revealed by 16S rRNA Gene Libraries

  • Zhang, Han-Bo;Shi, Wen;Yang, Ming-Xia;Sha, Tao;Zhao, Zhi-Wei
    • Journal of Microbiology
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    • v.45 no.6
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    • pp.479-484
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    • 2007
  • Bacterial communities at 10 cm, 100 cm, and 200 cm depths in a 100-year-old lead-zinc tailing heap were evaluated by constructing 16S rRNA gene libraries. In total, 98 operational taxonomic units (OTUs) were identified from 193 clones at a 3% sequence difference level. The OTU number and species richness decreased with the depth. Species composition was significantly different between the three libraries. Fifty-seven percent of the examined clones were Acidobacteria and 27% belonged to Proteobacteria. Other sequences included Chloroflexi, Firmicutes, Chlamydiae, Actinobacteria, Gemmatimonadetes, Nitrospira, and three unclassified OTUs. Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, Firmicutes, and Actinobacteria were mainly distributed in the rhizosphere of naturally colonizing plants; however, Deltaproteobacteria, Acidobacteria, and Chloroflexi tended to inhabit the deeper tailings (below the 100 cm-depth).

Diversity of Leuconostocs on Garlic Surface, an Extreme Environment

  • KIM, MYUNG HEE;SUN TAEK SHIM;YOUN SOON KIM;KYU HANG KYUNG
    • Journal of Microbiology and Biotechnology
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    • v.12 no.3
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    • pp.497-502
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    • 2002
  • Thirty-nine strains of Leuconostocs found to be tolerant to $10\%$ or more garlic were selected for further identification, by comparing their whole-cell protein pattern, 16S rRNA gene (first 530 bases) sequence, cellular fatty acid composition, and carbon source metabolism. Two isolates were Identified as Leuconostoc mesenteroides and 32 others as Leuconostoc citreum. Five other strains belonging to a cluster could not be allocated to the existing species. 16S rRNA gene sequence and cellular fatty acid composition of the unidentified bacteria exhibited close similarity with Leuconostoc argentinum. The unidentified isolates were not allocated to L. argentinum, because they formed polysaccharide from sucrose, while L. argentinum strains do not. Leuconostocs tolerant to high concentration of garlic were found predominantly on garlic surface, an extreme environment which is unfit for most of other microorganisms.

Cloning and Sequencing of Resistance Determinants to Aminoglycoside Antibiotics from Sterptoalloteichus hindustanus ATCC 31219 (Streptoalloteichus hindustanus ATCC 31219로부터 아미노글라이코사이드계 항생제에 내성을 지정하는 유전자의 클로닝 및 염기서열 결정)

  • Kim, Jong-Woo;Han, Jae-Jin;Choi, Young-Nae;Eom, Joon-Ho;Yoon, Sung-Joon;Hyun, Chang-Gu;Suh, Joo-Won
    • Microbiology and Biotechnology Letters
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    • v.23 no.4
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    • pp.384-389
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    • 1995
  • Streptoalloteichus hindustanus ATCC 31219, a nebramycin complex producer, is similar to Streptomyeces tenebrarius in a viewpoint of resistance to a wide range of aminoglycoside antibiotics. S. tenebrarius has resistance mechanisms of 16s rRNA methylation and aminogycoside modification. However, it is not known whether resistance mechanisms of Stall. hindustanus are the same as in S. tenebrarius. Therefore, we have tried to isolate resistance determinants from Stall. hindustanus. Two different types of aminoglycoside resistance determinants were isolated from Stall. hindustanus and expressed in Streptomyces lividans TK24. The apramycin resistance gene (amr) and the tobramycin resistance gene (tmr) isolated from Stall. hindustanus showed broad resistance spectrum against a dozen of aminoglycoside antibiotics. The complete nucleotide sequences of apramycin resistance gene (amr) were determined. The deduced amino acid sequence of the amr gene of Stall hindustanus ATCC 31219 showed extensive sequence homology to the 16s rRNA methylase gene (kamB) of S. tenebrarius.

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Bacterial Diversity and Distribution of Cultivable Bacteria Isolated from Dokdo Island (독도 주변의 해수에서 분리한 세균의 다양성과 군집구조 분석)

  • Sung, Hye-Ri;Ghim, Sa-Youl
    • Microbiology and Biotechnology Letters
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    • v.38 no.3
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    • pp.263-272
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    • 2010
  • One hundred sixty three strains showing different colony morphological characteristics on different concentration of marine agar (MA) plates were isolated from ambient seawater near Dokdo island. Bacterial diversity and distributions were studied by phylogenetic analysis of the partial 16S rRNA gene sequences. One hundred sixty three strains were partially sequenced and analyzed phylogenetically. They were composed of 5 phyla, of which gamma-proteobacteria (58%), alpha-proteobacteria (20%), bacteriodetes (16%) were predominant. They were affiliated with 90 species. The 16S rRNA sequence similarity of the isolates was in 93.3 to 100 % range to reported sequence data. Thirty six isolates of among them were assumed to be novel species candidates based on similarity analysis of the 16S rRNA gene sequences. Overall, Proteobacteria and Bacteriodetes of the Dokdo coastal sea water showed a high diversity.

Use of 16S-23S rRNA Intergenic Spacer Region for Species-specific Primer Developed of Vibrio Ichthyoenteri (16S-23S rRNA Intergenic Spacer Region을 이용한 Vibrio ichthyoenteri Species-specific Primer 개발)

  • Moon Young-Gun;Heo Moon-Soo
    • Korean Journal of Microbiology
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    • v.41 no.2
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    • pp.117-124
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    • 2005
  • Two bacterial isolates obtained from rotifer and diseased olive flounder larvae, Paralichthys olivaceus, were identified as Vibrio ichthyoenteri based on the results of phenotypic characterization. In an attempt to develop rapid PCR method for the detection of V. ichthyoenteri, we examined the 16S-23S rRNA intergenic spacer region(ISR) of V. ichthyoenteri and developed species-specific primer for V. ichthyoenteri. Analysis of the ISR sequences showed that V. ichthyoenteri contains one type of polymorphic ISRs. The size of ISRs was 348 bp length and did not contain tRNA genes. Mutiple alignment of representative sequences from different V. species revealed several domains of high sequence variability, and allowed to design species-specific primer for detection of V. ichthyoenteri. The specificity of the primer was examined using genomic DNA prepared from 19 different V. species, isolated 18group Vibrio species and most similar sequence of other known Vibrio species. The results showed that the PCR reaction using species-specific primer designed in this study can be used to detect V. ichthyoenteri.

Bacterial Diversity in a Korean Traditional Soybean Fermented Foods (Doenjang and Ganjang) by 16S rRNA Gene Sequence Analysis

  • Cho, Kye-Man;Seo, Weon-Taek
    • Food Science and Biotechnology
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    • v.16 no.2
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    • pp.320-324
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    • 2007
  • The bacterial diversity in Korean soybean-fermented foods was investigated using a PCR-based approach. 16S rRNA sequences were amplified and cloned from two different soybean-fermented foods such as doenjang (soybean paste), and ganjang (soybean sauce). Staphylococcus equorum (60.6%), Tetragenococcus halophila (21.2%), Leuconostoc mesenteroides (9.1%), Lactobacillus sakei (6.1%), and Bacillus subtilis (3.0%) were detected among clones isolated from soybean paste samples and Halanaerobium sp. (37.5%), Halanaerobium fermentans (37.5%), T. halophila (12.5%), Staphylococcus sp. (6.3%), S. equorum (3.1%), and B. subtilis (3.1%) were detected among clones isolated from soybean sauce. Our approach revealed different bacterial distributions and diversity from those previously obtained using culture-dependent methods.

Prevalence and molecular characteristics of 16s rRNA methylase gene rmtB in amikacin resistant Escherichia coli isolated from South Korea

  • Belaynehe, Kuastros Mekonnen;Won, Ho Geun;Yoon, In Joong;Yoo, Han Sang
    • Korean Journal of Veterinary Research
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    • v.59 no.3
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    • pp.157-160
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    • 2019
  • The production of rmtB-encoded 16S rRNA methylases has emerged as a novel mechanism promoting high-level resistance toward aminoglycosides in Gram-negative bacteria. Between 2015 and 2017, 636 distinct commensal Escherichia (E.) coli isolates were collected from different farms in South Korea to determine the prevalence and molecular characteristics of rmtB. The positive rates of rmtB between all the isolates and amikacin-resistant isolates were 1.1 and 100%, respectively. High-level aminoglycoside resistance could be transferred by conjugation from rmtB-positive donors to higher amikacin-resistance efficacies. This is the first report of 16S rRNA methylase-encoding genes in E. coli isolated from food-producing animals in Korea.