• Title/Summary/Keyword: RNA, Ribosomal, 16S

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Phylogeny of the subfamily Salmoninae distributed in Korea based upon nucleotide sequences of mitochondrial ribosomal RNA genes (미토콘드리아 ribosomal RNA 유전자 염기서열분석에 의한 한국산 연어아과 어류의 유전적 계통도)

  • LEE Heui-Jung;PARK Jung-Youn;LEE Jeong-Ho;MIN Kwang-Sik;JEON Im Gi;YOO Mi-Ae;LEE Won-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.33 no.2
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    • pp.103-109
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    • 2000
  • Complete senuences of the mitochondrial rRNA Benes were determined among six salmonines in Korean Waters (Brachpmystax lenok, Onoorhpchus keta, O. masou mason, O. mason ishikawae, O. mykiss, and albino mutant of O. mykiss). The purposes of this study were to provide the basic information on levels of mtDNA polymorphism among these species for genetic characterization; discuss phylogentic relationships among three Oncorhynchus sepecies; demonstrate the utility of rRNA gene sequence data as a genetic marker for disringuishinf among Korean salmonines. PCR/direct sequencing data indicated the following consistent results; 1) 12S rRNA genes was 945 bases long in Oncorhynchus species, and 946 bases in B. lenot including one insertion. 2) Of sequence variation in mitochondrial rRNA regions, transitional substitutions were superior to transversion. 3) The significant differences were not shown in the intraspecific variation values in these gene regions. The percentage sequence divergence values were ranged from $0.066 to 0.212{\%}$. 4) The interspecific divergences were greater than the intraspecific variation. Nevertheless, ribosomal RMh genes were more conserved among species than the other mitochondrial genes, and they showed potentiality as an intergenic marker for systematics. In addition, phylogenetic trees, constructed from this data, supported that cherry salmon was closer to chum salmon than to rainbow trout, and that lenok was most distantly related species in six salmonid species.

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Detection of Clostridium difficile by Loop-Mediated Isothermal Amplification (등온증폭법을 이용한 Clostridium difficile 검출)

  • In, Ye-Won;Ha, Su-Jeong;Yang, Seung-Kuk;Oh, Se-Wook
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.9
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    • pp.1326-1330
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    • 2012
  • This study was conducted to develop a loop-mediated isothermal amplification (LAMP) method for the detection of Clostridium difficile. The tested target gene was 16S ribosomal RNA. Five different LAMP primer sets were designed, and LAMP was performed. All primer sets targeting the 16S rRNA gene (BIP, FIP, B3, F3, LF, PF) were determined as positive in tcdA-positive, tcdB-postive ($A^+B^+$) and tcdA-negative, tcdB-negative ($A^-B^-$) Clostridium difficile strains. As the LAMP reaction took less than 80 min and did not require expensive machine such as thermocycler, it can be used as a rapid and simple detection method for foodborne pathogens.

Seasonal Variation of Bacterial Community in the Seawater of Gwangyang Bay Estimated by Amplified Ribosomal DNA Restriction Analysis (Amplified Ribosomal DNA Restriction Analysis를 이용한 광양만 해수의 세균 군집의 계절적 변화)

  • Ramos, Sonny Cachero;Hwang, Yeoung Min;Lee, Ji Hee;Baik, Keun Sik;Seong, Chi Nam
    • Journal of Life Science
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    • v.23 no.6
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    • pp.770-778
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    • 2013
  • To determine the seasonal variation of bacterial community in the seawater of Gwangyang Bay, three hundred thirty six bacterial strains were isolated on February, May, July and October 2011. Amplified Ribosomal DNA Restriction Analysis (ARDRA) was used to construct the phylotyes of the isolates using the restriction endonuclease, Hae III. Diversity indices of ARDRA patterns were calculated. One hundred and one phylotypes including 40 unique pylotypes were found at the 80% similarity level. Partial 16S rRNA genes of one hundred thirty nine strains representing each phylotypes were sequenced and compared. Bacterial community composed of 4 different phyla which include Proteobacteria, Actinobacteria, Bacteroidetes and Firmicutes. Proteobacteria was the prevailing phylum in all seasons, followed by Bacteroidetes in winter, spring and autumn while Actinobacteria in summer. At the family level, Flavobacteriaceae dominated in winter and spring and Pseudoalteromonadaceae did in summer and autumn. Genera Altererythrobacter, Loktanella, Pseudoalteromonas and Vibrio were encountered in all seasons. The most diverse bacterial community was found in autumn followed by the order of spring, winter and summer.

Functional Analysis of the Residues C770 and G771 of E. coli 16S rRNA Implicated in Forming the Intersubunit Bridge B2c of the Ribosome

  • Kim, Hong-Man;Yeom, Ji-Hyun;Ha, Hye-Jung;Kim, Jong-Myung;Lee, Kang-Seok
    • Journal of Microbiology and Biotechnology
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    • v.17 no.7
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    • pp.1204-1207
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    • 2007
  • Structural analyses have shown that nucleotides at the positions 770 and 771 of Escherichia coli 16S rRNA are implicated in forming one of highly conserved intersubunit bridges of the ribosome, B2c. To examine a functional role of these residues, base substitutions were introduced at these positions and mutant ribosomes were analyzed for their protein synthesis ability using a specialized ribosome system. The results showed requirement of a pyrimidine at the position 770 for ribosome function regardless of the nucleotide identity at the position 771. Sucrose gradient profiles of ribosomes revealed that the loss of protein-synthesis ability of mutant ribosome bearing a base substitution from C to G at the position 770 stems from its inability to form 70S ribosomes. These findings indicate involvement of nucleotide at the position 770, not 771, in ribosomal subunit association and provide a useful rRNA mutation that can be used as a target to investigate the physical interaction between 16S and 23S rRNA.

Genetic Analysis of a Structural Motif Within the Conserved 530 Stem-Loop of Escherichia coli 16S rRNA

  • Szatkiewicz Jin P.;Cho Hyun-Dae;Ryou Sang-Mi;Kim Jong-Myung;Cunningham Philip R.;Lee Kang-Seok
    • Journal of Microbiology and Biotechnology
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    • v.16 no.4
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    • pp.569-575
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    • 2006
  • The 530 stem-loop is a 46 nucleotide stem-loop structure found in all small-subunit ribosomal RNAs. Phylogenetic and mutational studies by others suggest the requirement for Watson-Crick interactions between the nucleotides 505-507 and 524-526 (530 pseudoknot), which are highly conserved. To examine the nature and functional significance of these interactions, a random mutagenesis experiment was conducted in which the nucleotides in the proposed pseudoknot were simultaneously mutated and functional mutants were selected and analyzed. Genetic analysis revealed that the particular nucleotide present at each position except 524 was not exclusively critical to the selection of functional mutants. It also indicated that basepairing interactions between the positions 505-507 and 524-526 were required for ribosomal function, and much weaker base-pairing interactions than those of the wild-type also allowed high ribosomal function. Our results support the hypothesis that the 530 pseudoknot structure may undergo a 'conformational switch' between folded and unfolded states during certain stages of the protein synthesis process by interacting with other ligands present in its environment.

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.

Bacterial Diversity in the Mud Flat of Sunchon Bay, Chunnam Provice, by 16S rRNA Gene Analysis (16S rRNA 유전자 분석에 의한 전남 순천만 갯벌의 세균 다양성)

  • 이명숙;홍순규;이동훈;배경숙
    • Korean Journal of Microbiology
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    • v.37 no.2
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    • pp.137-144
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    • 2001
  • In order to investigate the diversity of bacterial community in the mud flat of Sunchon Bay, Chunnam province, diversity of amplified 16S rDNA was examined. Total DNA was extracted from sediment soils and 16S rDNAs were amplified using PCR primers based on the universally conserved sequences in bacteria. Clonal libraries were constructed and 111 clones were examined by amplified rDNA restriction analysis (ARDRA) using HaeIII. Clones were clustered based on restriction patterns using computer program, GelCompar II. One hundred different RFLP types were detected from 111 clones. The 20 clones were selected and sequenced according to dendrograms derived from ARDRA, to cover most of the bacterial diversity in the clone libraries. None of the clones were identical to any representatives in the Ribosomal Database Project small subunit RNA databases and GenBank. All sequences showed between 77 and 96.8% similarity to the known 16s rRNA sequence from cultured organisms. The 20 clones sequenced fell into seven major lineages of the domain Bacteria: alpha-, delta-, gamma-Proteobacteria, low G+C Gram positive bacteria, high G+C Gram positive bacteria, Sphingobacteria (Cytophaga) and Cyanobacteria (chloroplast). Among the clones, the Proteobacteria were dominant.

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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.

Complete genome sequence of Gordonia sp. MMS17-SY073, a soil actinobacterium (토양 방선균인 Gordonia sp. MMS17-SY073 균주의 유전체 분석)

  • Kim, Yeong Seok;Kim, Seung Bum
    • Korean Journal of Microbiology
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    • v.55 no.3
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    • pp.303-305
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    • 2019
  • An actinobacterial strain designated Gordonia sp. MMS17-SY073 (=KCTC 49257) was isolated from a coastal soil of an island, and its complete genome was analyzed. A single contig consisting of 5,962,176 bp with the G + C content of 67.4% was obtained, and the annotation resulted in 5,201 protein-coding genes, 6 rRNA genes and 45 tRNA genes. Strain MMS17-SY073 was closest to the type strain of Gordonia soli based on the 16S rRNA gene sequence comparison, sharing 98.5% sequence similarity. A number of biosynthetic gene clusters for secondary metabolites, non-ribosomal peptide synthetase types in particular, could be identified from the genome.