• Title/Summary/Keyword: 16S rRNA genes

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Differentiation of Phytoplasmas Infecting Zizyphus jujuba and Paulownia coreana Using PCR-RELP

  • Han, Mu-Seok;Noh, Eun-Woon;Yun, Jeong-Koo
    • The Plant Pathology Journal
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    • v.17 no.4
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    • pp.189-193
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    • 2001
  • The relationships between the phytoplasmas infecting Zizyphus jujuba and Paulownia coreana were investigated by PCR-RELP. The 16S rRNA genes of the phytoplasmas were analyzed and compared with each other after PCR amplification. The amplified bands 1.4 kb in size were analyzed by both restriction digestion and sequencing after cloning into a plasmid vector. In some cases, two different kinds of inserts were observed in the isolates that originated from a single plant. However, many of them appeared to be the amplification products of chloroplastic 16S rRNA gene of host plants. The phytoplasma gene could be differentiated from the chloroplastic gene by restriction digestion of the plasmids carrying the amplification products. Only the recombinant plasmids carrying phytoplasma 16S rRNA gene produced a 1.4 kb band when digested with the enzyme BanII. Of the 52 recombinant plasmids analyzed, 42 appeared to contain inserts that originated from the chloroplastic 16S rRNA gene of the host plants. No variation was detected among 16S rRNA gene of nine phytoplasma isolates infecting Z. jujuba. However, the phytoplasmas infecting Z. jujuba were different from that infecting P. coreana.

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Genetic Identification of the Kimchi Strain Using PCR-based PepN and 16S rRNA Gene Sequence (PepN과 16S rRNA Gene Sequence 및 PCR 방법을 이용한 김치 젖산균의 동정)

  • Lee, Myung-Ki;Park, Wan-Soo;Lee, Byong-H.
    • Korean Journal of Food Science and Technology
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    • v.32 no.6
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    • pp.1331-1335
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    • 2000
  • The WL6 strain isolated from Kimchi could not be made scientific name because it was identified as three species, i.e., Leuconostoc mesenternides ssp cremoris, Leu. mesenteroides ssp. dextranicum or Lactobacillus bifermentans when it was tested by API kit or Biolog system methods. The unidentifiable WL6 strain was finally reclassified as Lactobacillus bifermentans by genetic identification using two PCR-based specific sequence primer sets which were originated from homologous pepN and 16S rRNA genes.

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Molecular Characterization and Prevalence of 16S Ribosomal RNA Methylase Producing Bacteria in Amikacin Resistant Gram-negative Bacilli Isolated from Clinical Specimens

  • Shin, Kyung-A;Hwang, Seock-Yeon;Hong, Seung-Bok
    • Biomedical Science Letters
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    • v.18 no.3
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    • pp.299-306
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    • 2012
  • Recently, the prevalence of 16S rRNA methylase conferring high-level resistance to aminoglycosides has been increasing in Gram-negative bacilli globally. We determined the prevalence and genotype of these methylase-producing bacteria, and characterized the co-resistance to ${\beta}$-lactam antibiotics and quinolone in Gram-negative clinical isolates collected in 2010 at a hospital in Korea. Among 65 amikacin-resistant isolates screened from 864 Gram-negative bacilli (GNB), 16S rRNA methylase genes were detected from 49 isolates, including Acinetobacter baumannii (43), Klebsiella pneumoniae (2), Proteus mirabilis (2) and Serratia marcescens (1), Empedobacter brevis (1). All of the 16S rRNA methylase genotype was armA and no variant sequences of amplified PCR products for armA were noted. The 16S rRNA methylase producing bacteria showed much higher resistance to aminoglycoside for Enterobacteriaceae and glucose non-fermenting (NF)-GNB and to imipenem for glucose NF-GNB, than the non-producing isolates. All of the 16S rRNA methylase producing Enterobacteriaceae had the extended-spectrum-${\beta}$-lactamase. In addition, two K. pneumoniae concurrently produced both plasmid-mediated AmpC ${\beta}$-lactamase and qnrB gene. All of the amikacin-resistant A. baumannii (43) co-harbored armA 16S rRNA methylase and $bla_{OXA-23}$ carbapenemase. In conclusion, 16S rRNA methylase producing bacteria were very prevalent among GNB in South Korea, and were commonly associated with co-resistance, including carbapenem and quinolone.

Emergence of CTX-M-15 Extended Spectrum β-lactamase and ArmA-Producing Enterobacter cloacae (CTX-M-15형 Extended Spectrum β-lactamase와 ArmA 동시 생성 Enterobacter cloacae의 출현)

  • Sung, Ji-Youn
    • Journal of Digital Convergence
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    • v.13 no.12
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    • pp.313-318
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    • 2015
  • We investigated the prevalence of extended spectrum ${\beta}$-lactamase (ESBL) genes and 16S rRNA methyltransferase genes to study antimicrobial resistance mechanisms of Enterobacter cloacae strains isolated from a university hospital in the Chungcheong province of Korea. Eight of the bacteria strains involved in this study contained CTX-M-15 type ESBL. Among 8 strains harboring the ESBL gene, 3 strains also harbored armA gene. The three isolates showed resistance to antimicrobial agents belonged to third cephalosporin, aminoglycoside, and fluoroquinolones. Furthermore, interspecies plasmid transfer of the antimicrobial resistant genes may induced horizontal spreading of the genes and emergence of multidrug resistant bacteria. Therefore, surveillance for existence of antimicrobial resistance determinants is important to prevent distribution of antimicrobial resistant strains.

Simple and Rapid Detection of Vancomycin-Resistance Gene from Enterococci by Loop-Mediated Isothermal Amplification

  • Baek, Yun Hee;Hong, Seung Bok;Shin, Kyeong Seob
    • Biomedical Science Letters
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    • v.26 no.3
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    • pp.149-156
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    • 2020
  • We developed a simple and rapid method for detecting vancomycin resistance genes, such as vanA and vanB, using loop-mediated isothermal amplification (LAMP). To identify not only vancomycin resistance genes, but also the genus Enterococcus, primers were designed for vanA, vanB, and 16S rRNA. Screening for vancomycin susceptibility in Enterococcus was performed using Etest (bioMérieux Inc). The results of the LAMP assay were compared to those of real-time RT-PCR. The optimal conditions for the LAMP assay were 65℃ for 60 min. The detection limits of the LAMP assay for vanA, and vanB were 2 × 102 copies/reaction. Compared to RT-PCR, the sensitivities and specificities of LAMP for 16S rRNA, vanA, and vanB were 100/100%, 100/100%, and 100/100%, respectively. The vanA genotype-vanB phenotype accounted for 57.5% (46/80) of the vancomycin-resistant Enterococci samples collected from 2016 to 2019. In conclusion, the LAMP assay developed in this study showed high sensitivity and specificity for vancomycin-resistant genes. Moreover, due to the simplicity and rapidity of the LAMP assay, its use can be very useful in clinical microbiology laboratories.

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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Assessment of the gastrointestinal microbiota using 16S ribosomal RNA gene amplicon sequencing in ruminant nutrition

  • Minseok Kim
    • Animal Bioscience
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    • v.36 no.2_spc
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    • pp.364-373
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    • 2023
  • The gastrointestinal (GI) tract of ruminants contains diverse microbes that ferment various feeds ingested by animals to produce various fermentation products, such as volatile fatty acids. Fermentation products can affect animal performance, health, and well-being. Within the GI microbes, the ruminal microbes are highly diverse, greatly contribute to fermentation, and are the most important in ruminant nutrition. Although traditional cultivation methods provided knowledge of the metabolism of GI microbes, most of the GI microbes could not be cultured on standard culture media. By contrast, amplicon sequencing of 16S rRNA genes can be used to detect unculturable microbes. Using this approach, ruminant nutritionists and microbiologists have conducted a plethora of nutritional studies, many including dietary interventions, to improve fermentation efficiency and nutrient utilization, which has greatly expanded knowledge of the GI microbiota. This review addresses the GI content sampling method, 16S rRNA gene amplicon sequencing, and bioinformatics analysis and then discusses recent studies on the various factors, such as diet, breed, gender, animal performance, and heat stress, that influence the GI microbiota and thereby ruminant nutrition.

Morphological and Molecular Analyses of $Anabaena$ $variabilis$ and $Trichormus$ $variabilis$ (Cyanobacteria) from Korea

  • Choi, Gang-Guk;Yoon, Sook-Kyung;Kim, Hee-Sik;Ahn, Chi-Yong;Oh, Hee-Mock
    • Korean Journal of Environmental Biology
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    • v.30 no.1
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    • pp.54-63
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    • 2012
  • This study characterizes three $Anabaena$ strains and 5 $Trichormus$ strains isolated from Korean waters and 3 $Anabaena$ $flos-aquae$ strains procured from the UTEX based on morphological features and molecular analyses. The $Anabaena$ and $Trichormus$ isolates were morphologically assigned to $A.$ $variabilis$ K$\ddot{u}$tzing and $T.$ $variabilis$(K$\ddot{u}$tzing ex Bornet et Flahault) Kom$\acute{a}$rek et Anagnostidis, respectively. The $Anabaena$ and $Trichormus$ strains differed significantly in the mean length of their vegetative cells. The 16S rRNA genes from the $Anabaena$ strains showed a 100% identity to that from $A.$ $variabilis$ ATCC 29413, while the 16S rRNA genes from the $Trichormus$ strains showed a 99.9% identity to that from $T.$ $variabilis$ GREIFSWALD. The overall topology was in agreement for the 16S rRNA gene and $cpcBA$-IGS trees in the both tree-constructing methods. In a neighbor-joining tree based on the 16S rRNA gene, the 3 $Anabaena$ strains were asso-ciated with $A.$ $variabilis$, the 5 $Trichormus$ strains with $T.$ $variabilis$, and the 3 $Anabaena$ (UTEX) strains were with $Nostoc$. To date, this is the first report on $A.$ $variabilis$ and $T.$ $variabilis$ strains originating from Korea.

Application of Next Generation Sequencing to Investigate Microbiome in the Livestock Sector (Next Generation Sequencing을 통한 미생물 군집 분석의 축산분야 활용)

  • Kim, Minseok;Baek, Youlchang;Oh, Young Kyoon
    • Journal of Animal Environmental Science
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    • v.21 no.3
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    • pp.93-98
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    • 2015
  • The objective of this study was to review application of next-generation sequencing (NGS) to investigate microbiome in the livestock sector. Since the 16S rRNA gene is used as a phylogenetic marker, unculturable members of microbiome in nature or managed environments have been investigated using the NGS technique based on 16S rRNA genes. However, few NGS studies have been conducted to investigate microbiome in the livestock sector. The 16S rRNA gene sequences obtained from NGS are classified to microbial taxa against the 16S rRNA gene reference database such as RDP, Greengenes and Silva databases. The sequences also are clustered into species-level OTUs at 97% sequence similarity. Microbiome similarity among treatment groups is visualized using principal coordinates analysis, while microbiome shared among treatment groups is visualized using a venn diagram. The use of the NGS technique will contribute to elucidating roles of microbiome in the livestock sector.

Phylogenetic Relationship and DNA Polymorphism of Boleophthalmus pectinirostris and Scartelaos gigas (Teleostei: Gobiidae) of Korea (한국산 짱뚱어(Boleophthalmus pectinirostris)와 남방짱뚱어(Scartelaos gigas) (Gobiidae)의 분자유전학적 계통연관과 DNA 다형화)

  • Choi, Ki Ho;Chung, Ee Yung;Park, Gab Man
    • Korean Journal of Ichthyology
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    • v.25 no.3
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    • pp.149-156
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    • 2013
  • Phylogenetic relationships and DNA polymorphism among local populations of two Korean gobiidae species: Boleophthalmus pectinirostris and Scartelaos gigas were investigated based on 12S and 16S mitochondrial DNA and mitochondrial cytochrome b DNA sequences. DNA polymorphisms of B. pectinirostris between Suncheon and Gunsan populations were 100% identity from 434 bp segment of 12S rRNA gene and from 444 bp segment of mitochondrial cytochrome b genes, and 99.6% (2 bp different) identity from 484 bp segments of 16S rRNA genes. These results indicated the long period of geographic isolation between two populations of B. pectinirostris in Korea caused such high degrees of DNA polymorphisms. Based on the phylogenetic tree constructed from the two gobiid species in Korea, two genetically distinct groups of B. pectinirostris and S. gigas groups were recognized.