• Title/Summary/Keyword: 16S-rRNA

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Taxonomic characteristics of novel Flavobacteriumsp. B1 from a freshwater pond

  • Bae, Young-Min
    • Journal of the Korean Applied Science and Technology
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    • v.39 no.5
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    • pp.605-613
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    • 2022
  • The genus Flavobacterium, type genus of the family Flavobacteriaceae and a member of the phylum Bacteriodetes includes gram-negative and yellow-pigmented rods. Those bacteria have been isolated from various environments of the earth. A yellow-pigmented, gram-negative rod was isolated from a pond in the campus of the Changwon University, Changwon, Kyeongnam and designated as strain B1. Strain B1 was further analyzed physiologically, biochemically and phylogenetically, and concluded to be a member of genus Flavobacterium. BLAST search of the 16S rRNA gene sequence of strain B1 shows homology no higher than 99.0% with those sequences of other bacteria. The major fatty acids of strain B1 are iso-C15:0 (19.6%), summed feature 3(C16:1 ω7c and/or C16:1 ω6c, 16.1%), iso-C17:0 3OH(10.2%), iso-C15:0 3OH(8.4%) and iso-C15:1 G(6.6%) showing significant differences in fatty acid compositions between strain B1 and the other known Flavobacterium species. DNA sequence of 16S rRNA gene of strain B1 was deposited in genbank under accession number OP060681.

Characterization of beer-spoilage microorganism and its rapid detection by specific PCR primer (맥주오염미생물의 동정과 specific PCR primer의한 신속한 검출 방법)

  • Lee, Taek-In;Choi, Shin-Geon
    • Journal of Industrial Technology
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    • v.28 no.A
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    • pp.141-147
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    • 2008
  • Several contaminated bacteria such as Lactobacillus brevis and Pediococcus damnosus in beer production cause beer spoilage by producing off flavours and turbidity. Detection of these organisms is complicated by the strict anaerobic conditions and lengthy incubation times required for their cultivation, consequently there is a need for more rapid detection methods. Recently, two contaminated strains were isolated from vessel of beer production and identified as Lactobacillus species by API kit identificaton as well as 16S-23S ITS sequencing analyses. Two isolated strains were named as Lactobacillus sp. HLA1 and Lactobacillus HLB2, respectively. A polymerase chain reaction (PCR) method was developed for the rapid and specific detection of Lactobacillus sp.. Two sets of primer pairs (HLA1-F/HLA1-R and HLB2-F/HLB2-R) were designed for the amplification of a 1576 base pair (bp) fragment of the HLA1 16S-23S rRNA gene and 1888 bp fragement of the HLB2 16S-23S rRNA. Amplified PCR products were highly specific to detect corresponding bacteria when other contaminated strains were used as PCR templates. However, detection of both strains were limited when $100{\mu}{\ell}$ of cultured samples were mixed with $100m{\ell}$ of beer sample in arbitrary manner. The sensitivity of the assay still needs to be improved for direct detection of the small amounts of bacteria present in beer.

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Comparison between DNA- and cDNA-based gut microbial community analyses using 16S rRNA gene sequences (16S rRNA 유전자 서열 분석을 이용한 DNA 및 cDNA 기반 장내 미생물 군집 분석의 비교)

  • Jo, Hyejun;Hong, Jiwan;Unno, Tatsuya
    • Korean Journal of Microbiology
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    • v.55 no.3
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    • pp.220-225
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    • 2019
  • Studies based on microbial community analyses have increased in the recent decade since the development of next generation sequencing technology. Associations of gut microbiota with host's health are one of the major outcomes of microbial ecology filed. The major approach for microbial community analysis includes the sequencing of variable regions of 16S rRNA genes, which does not provide the information of bacterial activities. Here, we conducted RNA-based microbial community analysis and compared results obtained from DNA- and its cDNA-based microbial community analyses. Our results indicated that these two approaches differed in the ratio of Firmicutes and Bacteroidetes, known as an obesity indicator, as well as abundance of some key bacteria in gut metabolisms such as butyrate producers and probiotics strains. Therefore, cDNA-based microbial community may provide different insights regarding roles of gut microbiota compared to the previous studies where DNA-based microbial community analyses were performed.

Genetic Divergence and Phylogenetic Relationships among the Korean Fireflies, Hotaria papariensis, Luciola lateratis, and Pyrocoelia rufa(Coleoptera: Lampyridae), using Mitochondrial DNA Sequences (미토콘드리아 DNA의 염기서열을 이용한 파파리반딧불이, 애반딧불이 및 늦반딧불이 (딱정벌레목: 반딧불이과)의 유전적 분화 및 계통적 관련)

  • 김익수;이상철;배진식;진병래;김삼은;김종길;윤형주;양성렬;임수호
    • Korean journal of applied entomology
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    • v.39 no.4
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    • pp.211-226
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    • 2000
  • Genetic divergence and phylogenetic relationships among the major Korean fireflies (Hotaria papariensis, Luciola lateralis, and Pyrocoelia rufa) were studied. A portion of mitochondrial COI (403 bp) and 165 rRNA (490~504 bp) genes were sequenced, and the GenBank-registered, homologous 165 rRNA sequences of Japanese fireflies were compared (27 species of Lampyridae, one of Lycidae, and one of Rhgophthalmidae). Greatest DNA and/or amino acid sequence divergence was found when P rufa, belonging to Lampyrinae was compared with H. papariensis and L. lateralis, both belong-ing to Luciolinae, confirming the current taxonomic status of the species. In the PAUP and PHYLIP analyses with 165 rRNA data, grouping of the two geographic samples of H. papariensis with H. tsushimana validate the use of generic name, Hotaria. Nevertheless, lack of sister-group relationship of the two geographic samples of H. papariensis renders further investigation on this group . Although the Korean and Japanese L. lateralis formed a strong monophyletic group, a substantial genetic differentiation was detected between them (2.9% of 165 rRNA gene sequence divergence). Finally, the geographic samples of Korean p. rufa strongly formed a group with Japanese p. rufa, warranting the use of generic name, Pyrocoelia, but the genetic distance observed between the Cheju-Island individual and all others requires further investigation on this subject. Summarized, this study supports the current taxonomic status of the Korean fireflies in that each respectively formed a strong monophyletic group with its own species or genus.

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Molecular Systematics of the Tephritoidea (Insecta: Diptera): Phylogenetic Signal in 16S and 28S rDNAs for Inferring Relationships Among Families

  • Han, Ho-Yeon;Ro, Kyung-Eui;Choi, Deuk-Soo;Kim, Sam-Kyu
    • Animal cells and systems
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    • v.6 no.2
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    • pp.145-151
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    • 2002
  • Phylogenetic signal present in the mitochondrial 16S ribosomal RNA gene (16S rDNA) and the nuclear large subunit ribosomal RNA gene (28S rDNA) was explored to assess their utility in resolving family level relationships of the superfamily Tephritoidea. These two genes were chosen because they appear to evolve at different rates, and might contribute to resolve both shallow and deeper phylogenetic branches within a highly diversified group. For the 16S rDNA data set, the number of aligned sites was 1,258 bp, but 1,204 bp were used for analysis after excluding sites of ambiguous alignment. Among these 1,204 sites, 662 sites were variable and 450 sites were informative for parsimony analysis. For the 28S rDNA data set, the number of aligned sites was 1,102 bp, but 1,000 bp were used for analysis after excluding sites of ambiguous alignment. Among these 1000 sites, 235 sites were variable and 95 sites were informative for parsimony analysis. Our analyses suggest that: (1) while 16S rDNA is useful for resolving more recent phylogenetic divergences, 28S rDNA can be used to define much deeper phylogenetic branches; (2) the combined analysis of the 16S and 28S rDNAs enhances the overall resolution without losing phylogenetic signal from either single gene analysis; and (3) additional genes that evolve at intermediate rates between the 16S and 28S rDNAs are needed to further resolve relationships among the tephritoid families.

Specific Primer for Detection of Jujube Witches' Broom Phytoplasma Group (16SrV) in Korea

  • Han, Sang-Sub
    • The Plant Pathology Journal
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    • v.21 no.1
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    • pp.55-58
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    • 2005
  • In order to diagnose and differentiate jujube witches' broom (JWB) phytoplasma rapidly, oligonucleotide primer pair, 16Sr(V) F/R, for polymerase chain reactions (PCRs) was designed on the basis of 16S rRNA sequences of JWB phytoplasma. The PCR employing phytoplasma universal primer pair P1/P7 consistently amplified DNA in all tested phytoplasma isolates. But no phytoplasma DNA was detected from healthy jujube seedlings. The nested PCR, the primer pair 16S(V) F/R, about 460 bp fragment, amplified DNA in all tested JWB and related phytoplasmas including ligustrum witches' broom phytoplasma of the 16S rRNA group V, but no DNA amplification was detected from other phytoplasma strains such as groups 16SrI (Aster yellows) and 16SrXII (Stolbur group) in which mulberry dwarf phytoplasma and chrysanthemum witches' broom phytoplasma belong to, respectively. The same results were obtained from both Korean and Chinese isolates of JWB phytoplasma. Nested-PCR using phytoplasma universal primer pair P1/P7 and 16SrV group-specific primer pair 16S(V) F/R could detect group V phytoplasmas rapidly and easily, in particular JWB phytoplasma.

유전자보유 계통수를 이용한 Archaea와 Proteobacteria 분류

  • Lee, Dong-Geun;Lee, Jin-Ok;Lee, Jae-Hwa
    • 한국생물공학회:학술대회논문집
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    • 2003.04a
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    • pp.686-689
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    • 2003
  • A Gene content phylogenetic tree and a 16S rRNA based phylogenetic tree were compared for 9 Archaea and 15 Proteobacteria, whole-genome sequenced, by neighbor joining and bootstrap methods (n=1000). Ratio of conserved COG (clusters of orthologous groups of proteins) to ortholog revealed that they were within the range of 4.60% (Mezorhizobium loti) or 56.57% (Mycoplasma genitalium), The diversity of ratio meant the Possibility of searching for useful genes, as they possess peculiar genes. The gene content tree and the 16S rDNA tree showed coincidence and discordance in Archaea and Proteobacteria.

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Analysis of 16S-23S rRNA Intergenic Spacer Regions of Aeromonas veronii biogroup sobria and A. caviae (Aeromonas veronii biogroup sobria와 Aeromonas caviae의 16S-23S rRNA Intergenic Spacer Regions 분석)

  • 강동율;이훈구
    • Korean Journal of Microbiology
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    • v.36 no.3
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    • pp.173-180
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    • 2000
  • The intern1 spacer regions (ISR) between the 16s and 23s $1_RNA$ genes of Aeronzonus iwonii blogroupsobria and A. caviae were investigated by PCR fragment length typing and DNA sequencing. A. iwonii bv.sobria has a speciIic 16s-23s pattern of 2-4 fiagments ranging Goin 479-539 bp, with the exception of thespecies Aeron7onns cmiae, which has 3 fragments ranglog from 470-602 bp. In all of the.4 vei*onii bv. sobr,iaand A, caviae strains examined in this study, the 470-481bp Tragnent, designated TSR-1, invariably contained $tDNA^{uc(GAT)$ and $tDNA^{Ala(TGC)$ in contrast to ISR-2 (513-525 bp). ISR-3 (537-539 bp) and ISR-4 (568-602 bp)containing TEX>$tDNA^{Olu(ITC)$ A stretch of 20 nucleotides (178-197 bp) in the ISR-4 was conserved only wit11mA.caiiue, from which the A. caiiae specific primer, named prAC-F, was designed and used for PCR with aAcaviae coimnon reverse primer A PCR product of 450 bp was apparent alnong I , caiizne strains, but not ii1.4.ijeronii bv. sob~ia strains. The PCR product was oot detected t"-om strains belonging to A. hjili-o~~hila, Ebrio,aud the family Ef\ulcornertei,obncteriaceae. This study provides the first molecular tool for mdentifying the species 8.caviae.ing the species 8. caviae.

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Comparison of Culture-dependent and DGGE based Method for the Analysis of Marine Bacterial Community (배양법과 DGGE에 의한 해양세균 군집의 비교분석)

  • Kim, Mal-Nam;Bang, Hyo-Joo
    • Korean Journal of Environmental Biology
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    • v.24 no.4
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    • pp.307-313
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    • 2006
  • Seasonal variation of marine bacterial community was analyzed in the surface sea water collected from one of the stations locating at Tongyeoung coastal area, Korea. The results obtained by the culture method through identification with the VITEK Microbe ID system after pure culture in the selective medium were compared with those obtained by the DGGE based 16S rRNA PCR method. The composition of the marine bacterial community in the sea water samples harvested in September, 2004, November, 2004, January, 2005, May, 2005 and August, 2005 determined by the culture method showed 5, 5, 4, 6, and 10 strains respectively. Pseudomonas fluorescens and Acinetobacter lwoffii were detected in all seasons. The other strains were identified to be Pseudomonas stutzeri, Sphingomonas paucimobilis, Burkholderia mallei and Chryseobacterium indologenes. In contrast, the 16S rRNA PCR-DGGE method detected 10, 11, 6, 9 and 13 populations respectively in the same sea water samples and the strains were identified to be Acinetobacter lwoffii, Burkholderia mallei, Pseudomonas fluoresence, Actinobacillus ureae, Burkholderia sp., Pseudomonas stutzeri, Roseobacter sp., Vibrio parahaemolyticue, Sphingomonas paucimobilis and Rugeria algocolus. This results indicated that the DGGE based 16S rRNA PCR method was more efficient than the culture method for the grasp of the characteristics of the marine bacterial community.

Molecular Identification of the Bloom-forming Cyanobacterium Anabaena from North Han River System in Summer 2012 (북한강 수계 조류대발생 원인종 남조 Anabaena의 분자계통학적 검토)

  • Li, Zhun;Han, Myung-Soo;Hwang, Su-Ok;Byeon, Myeong-Seop;Hwang, Soon-Jin;Kim, Baik-Ho
    • Korean Journal of Ecology and Environment
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    • v.46 no.2
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    • pp.301-309
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    • 2013
  • Between May and August 2012, a massive cyanobacterial bloom with Anabaena has been occurred throughout the North Han River. Sampling was conducted at one station on each lake, L. Uham, L. Cheongpyung, and L. Paldang, where occurred a dense bloom, in 13 July. According to the microscopic examination, the blooms was dominated by one specific filamentous cyanobacterium Anabaena and other phytoplankton. Morphologically, previous literature proven that this Anabaena species is A. crassa (Lemmermann) Komark.-Legn. & Cronberg. However, identification of species in a mixed population is complicated due to limited morphological differences. Therefore, with live sample including trichome, akinete and heterocyst, the sequences of 16S rRNA gene of Anabaena isolates were cloned and analyzed, and three 16S rRNA gene sequences of 1188~1520 bp in length were obtained. It was shown from the homologous analysis results that the obtained 16S rRNA sequences were highly homologous to the relevant sequences of A. crassa in GenBank. The 16S rRNA sequences of 63 species were retrieved from GenBank, and the phylogenetic tree was constructed by using these sequences.