• Title/Summary/Keyword: 26S rDNA

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Karyotype Analysis and rDNA Physical Mapping in Rye (Secale cereale L.) (호밀(Secale cereale L.)의 핵형분석과 rDNA의 Physical Mapping)

  • Lee, Joon Soo;Seo, Bong Bo;Kim, Min
    • Korean Journal of Breeding Science
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    • v.42 no.2
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    • pp.163-168
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    • 2010
  • This study was carried out to determine the chromosomal localization of the 5S and 18S-26S ribosomal DNA(rDNA) genes by means of fluorescence in situ hybridization(FISH) techniques, and the constitutive heterochromatin detected by means of Gimsa C-banding technique in rye(Secale cereale L.). The somatic chromosomes number was 2n=14. The karyotype consists of four pairs of metacentrics(chromosomes 1, 2, 3, and 7) and three pairs of submetacentrics(chromosomes 4, 5, and 6). Secondary constrictions appeared in the short arm of chromosome 1. The 5S rDNA genes have been located on two pairs of chromosomes 1 and 5, and 18S-26S rDNAs genes have been located on one pair of chromosome 1. 5S rDNA genes were detected on the distal region of the secondary constrictions in nucleolus organizer regions(NOR) in chromosome 1, and other detected on the intercalary region in the short arm of chromosome 5.

Molecular Biological Identification of Bacteria in Middle Ear Effusion Using 16S rDNA Multiplex PCR (중이 삼출액 미생물의 16S rDNA 복합중합효소연쇄반응을 이용한 분자생물학적인 진단)

  • 이정구;이인숙;박지연;정상운;오충훈
    • Korean Journal of Microbiology
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    • v.39 no.1
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    • pp.36-39
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    • 2003
  • The rapid and reliable 16S rDNA multiplex polymerase chain reaction (PCR) assay was established to characterize bacterial etiologies of middle ear effusion. These etiologies included Haemophilus influenzae, Moraxella catarrhalis and Streptococcus pneumonia, which were detected in middle-ear effusion (MEE) samples taken from patient with otitis media. A total of 39 MEE samples were aspirated from 26 patients. DNA was extracted from MEE samples, and PCR was done with DNA extracts by using the common primers, which is localized at C4 region in the 16S rDNA gene of all bacterial species, and species-specific primers: (i) Haemophilus-specific primer, (ii) Moraxella- specific primer, and (iii) Streptococcus-specific primer. Among 39 samples tested, 24 (61.5%) were positive for H. influenzae, 10 (25.6%) were positive for M. catarrhalis, 3(7.7%) were positive for S. pneumonia, and 11 (28%) were negative for 165 rDNA multiplex PCR reaction. Nine samples (28.6%) exhibited a mixed infection and were positive for both H. infuenzae and M. catarrhalis. We suggested that 16S rDNA multiplex PCR is a useful method to identify rapidly for rapid identification of the pathogenic bacteria and characterization of bacterial etiologies of middle ear effusion.

Cytotaxonomical Study of the Chenopodium album and its Related Species in Korea (한국산 흰명아주와 근연종의 세포분류학적 연구)

  • Chung, Youngjae;l Kim, Muyeol;Lee, Byongsoon
    • Korean Journal of Plant Taxonomy
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    • v.41 no.4
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    • pp.324-328
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    • 2011
  • The purpose of this study was to analyze the interspecific relationships of Chenopodium album and its related taxa collected in Korea. The 18S-26S ribosomal DNA (45S rDNA) loci were detected directly on mitotic chromosomes by fluorescence in situ hybridization (FISH) and the chromosome numbers were examined using aceto-orcein methods. The chromosomal numbers of Chenopodium album var. album and C. album var. centrorubrum were 2n = 6x = 54, whereas for C. album var. stenophyllum, this number was 2n = 4x = 36. The basic chromosome number was x = 9. The biotin labeled 18S-26S rDNA probe exhibited eight yellow fluorescent signals on the metaphase chromosome of C. album var. album and var. centrorubrum respectively, while two yellow signals of C. album var. stenophyllum were noted. All of the signals on the chromosomes were located at the terminal regions. The chromosome number and FISH findings suggest that C. album var. centrorubrum is merged into var. album and that it is clearly distinguished from C. album var. stenophyllum.

The 18s rDNA Sequences of the Basidiocarps of Tricholoma matsutake in Korea (한국산 송이버섯에서의 18s ribosomal DNA 서열)

  • Lee, Sang-Sun;Hong, Sung-Woon
    • The Korean Journal of Mycology
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    • v.26 no.2 s.85
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    • pp.256-264
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    • 1998
  • The 18S rDNA sequences of Tricholoma matsutake (TM=T. caligatum var. nauseoum) collected in Korea were analyzed for the ectomycorrhizal fungi in the roots of Pinus densiflora. The 514 base pairs of rDNA region were synthesized by UF-5 and UR-6 primers, and double checked in the base pair. The sequence of four strains synthesized were all identical in this work, but different from those done by the previous workers. The basidiocarps collected in this work. were identified to T. matstake after searching the 18s rDNA by the BLAST in NCBI. Only several base pairs of 18S rDNA analyzed from other related basidiocarps were different from our analyses of 18S rDNA. The dendrogram were made based on the sequences of the 514 bp 18S rDNA by CLUSTAL-X alignment program. The groupings of the species at the level of genus in the dendrogram were well constructed.

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Analysis of Chromosome Composition of Gastrodia elata Blume by Fluorescent in situ Hybridization using rDNA and Telomeric Repeat Probes (rDNA와 말단소체 반복서열 탐침을 이용한 천마의 FISH 염색체 조성 분석)

  • Zhou, Hui Chao;Park, Eung Jun;Kim, Hyun Hee
    • Korean Journal of Medicinal Crop Science
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    • v.26 no.2
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    • pp.113-118
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    • 2018
  • Background: Gastrodia elata Blume is a saprophytic perennial plant in the Orchidaceae family, because of its agricultural and medicinal effectiveness, researchers focus on its genome and chemical components. However, cytogenetic information based on the chromosome structure and composition to construct chromosomal backbone for genome sequencing research and for the development and breeding of plants is very limited. Methods and Results: We determined the metaphase chromosome composition of the G. elata genome by fluorescence in situ hybridization (FISH) using 5S and 45S rDNAs and telomeric repeat probes. The nuclear genome of G. elata was organized into 2 n = 36, with relatively small ($2.71-5.50{\mu}m$)chromosomes that showed gradual decrease in size. Conglutination phenomenon was observed among the metaphase chromosomes, and it was distinguished from that in other plant metaphase chromosome spreads. One pair of signal was detected for each 5S and 45S rDNA in the pericentromeric region and interstitial region on the short arm of chromosomes 10 and 4, respectively, and telomeric DNA signals were detected in the terminal region of most chromosomes. Conclusions: To our knowledge, this is the first FISH chromosome composition result in G. elata and could be useful in more comprehensive molecular cytogenetic and genomic analyses as well as breeding programs of the medicinal plant G. elata.

Genetic identification of Aeromonas species using a housekeeping gene, rpoD, in cultured salmonid fishes in Gangwon-Do (강원도 양식 연어과 어류에서 분리된 에로모나스 종의 유전학적 동정)

  • Lim, Jongwon;Koo, Bonhyeong;Kim, Kwang Il;Jeong, Hyun Do;Hong, Suhee
    • Journal of fish pathology
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    • v.30 no.2
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    • pp.79-88
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    • 2017
  • At the present, fish farms are suffering a lot of economic losses due to infectious diseases caused by various pathogens including aeromonad. Aeromonad is ubiquitous bacteria that causes infectious diseases. At least 26 species in the genus Aeromonas have been reported to cause fatal infections not only in salmonid fishes, but also in other freshwater and seawater fishes. Molecular techniques based on nucleic acid sequences of 16S rDNA and housekeeping genes can be used to identify the Aeromonas species. In this study, The genus Aeromonas was isolated from salmonid fishes of sixteen fish farms in Gangwon-Do, Korea and phylogenetically identified based on the sequences of 16S rDNA and housekeeping genes for Aeromonad, i.e. RNA polymerase sigma factor ${\sigma}^{70}$ (rpoD) or DNA gyrase subunit B (gyrB). Consequently, 96 strains were collected from Atlantic salmon (Salmo salar), coho salmon (Oncorhynchus kisutch), masou salmon (Oncorhynchus masou) and rainbow trout (Oncorhynchus mykiss), and 36 isolates were identified as the genus Aeromonas by 16S rDNA analysis. Thirty six Aeromonad isolates were further analysed based on rpoD or gyrB gene sequences and found Aeromonas salmonicida (24 isolates), A. sobria (10 isolates), A. media (1 isolates) and A. popoffii (1 isolates), indicating that A. salmonicida is a main infectious bacteria in Salmonid fishes in Gangwon-Do. It was also proved that the phylogenetic identification of Aeromonas species based on the sequences of housekeeping gene is more precise than the 16S rDNA sequence.

DNA Sequencing and Phylogenetic Analysis of the 18S rRNA Gene of Atractylodes japonica Koidz and Analysis of Atractylon (삽주의 18S rRNA 유전자의 염기서열 결정, 계통분류학적 분석 및 atractylon 분석)

  • Bae, Young-Min
    • Korean Journal of Medicinal Crop Science
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    • v.17 no.1
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    • pp.26-32
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    • 2009
  • The region containing 18S rRNA gene, ITS 1 and part of the 5.8S rRNA gene of the Atractylodes japonica Koidz was amplified by PCR and the product cloned in a pBluescript SK II plasmid. DNA sequence of the cloned DNA was determined and submitted to the GenBank (accession number EU678363). Phylogenetic analysis of the ITS 1 DNA showed close similarity with the other plant species of the family Compositae. The extract of the plant materials of five different members of the family Compositae was analyzed by HPLC to detect atractylon. Extract of the A. japonica Koidz showed presence of significant amount of atractylon. However, noticeable amount of atractylon was not detected by the same analyses from the extracts of the other plants belonging to the family Compositae including Artemisia capillaris, Chrysantemum zawadskii, Eclipta prostrata or Taraxacum platycarpum.

Analysis of Bacterial Community Structure in Bulk Soil, Rhizosphere Soil, and Root Samples of Hot Pepper Plants Using FAME and 16S rDNA Clone Libraries

  • Kim, Jong-Shik;Kwon, Soon-Wo;Jordan, Fiona;Ryu, Jin-Chang
    • Journal of Microbiology and Biotechnology
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    • v.13 no.2
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    • pp.236-242
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    • 2003
  • A culture-independent and -dependent survey of the bacterial community structure in the rhizosphere and soil samples from hot pepper plants was conducted using 16S rDNA clone library and FAME analyses. Out of the 78 clones sequenced, 56% belonged to Proteobacteria, 4% to high G+C Gram- positive group, 3% to Cytophyga-Flexibacter-Bacreroides, and 32% could not be grouped with any known taxonomic division. Among the 127 FAME isolates identified, 66% belonged to low G+C Gram-positive bacteria (Baciilus spp.) and 26% to high G+C Gram-positive bacteria. In a cluster analysis, the results for both methods were found to be strikingly dissimilar. The current study is the first comparative study of FAME and 165 rDNA clonal analyses performed on the same set of soil, rhizosphere soil, and root samples.

Identification of the Orchid Mycorrhizal Fungi Isolated from the Roots of Korean Native Orchid

  • Lee, Sang-Sun;You, Jae-Hyung
    • Mycobiology
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    • v.28 no.1
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    • pp.17-26
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    • 2000
  • The orchid symbiotic fungi were isolated from the roots of Korean native orchid (Cymbidium goeringii) collected and Chinese orchid (C. sinense) obtained from greenhouses. They were identified as a species of Rhizoctonia, based on the sequences of 18r rDNA, the microscopic observations of mycelia, and the symbiotic relationships with commercial orchids. The isolate collected from Chinese orchids was revealed to be a species of Ceratobasidium endophytica, and to be different from the other isolates at the thickness of the mycelia stained in the root cells of Korean native orchids. The other isolates collected from the Korean native orchids were considered to be a species of Tulsanella repens (anamorphic: Epulorhiza repens) or its related one. The physiologic or microscopic variations were oftenly observed among them, but the tendency of grouping these in the 18s rDNA sequences were observed to be consistent with those of the localities collected. The further taxonomical segregating for Korean symbiotic fungi was not made because the information concerned were limited in this moment, but was recognized as based on the sequences of 18s DNA.

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The Diversity of Heterotrophic Bacteria Isolated from Intestine of Starfish(Asterias amurensis) by Analysis of 16S rDNA Sequence (16S rDNA염기서열에 의한 불가사리(Asterias amurensis) 장내에서 분리된 종속영양세균 군집의 다양성)

  • Choi, Gang-Guk;Lee, Oh-Hyung;Lee, Geon-Hyoung
    • The Korean Journal of Ecology
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    • v.26 no.6
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    • pp.307-312
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    • 2003
  • To study the diversity of heterotrophic bacteria isolated from intestine of starfish, Asterias amurensis, we collected starfishes from the coastal area near Jangheung-Gun, Jeollanam-Do, Korea during July, 2000. Population density and bacterial diversity in the intestine of starfish were measured. The results were as follows; The population densities of heterotrophic bacteria in the intestine of starfish were 8.65${\pm}$0.65${\times}10^3\;dfu\;g^{-1}$. Gram positive bacteria occupied 59% among 29 isolates. The community structure of dominant heterotrophic bacteria in the intestine of starfish consisted of Bacillaceae in the low G+C gram positive bacteria subphylum, Microbacteriaceae in the high G+C gram positive bacteria subphylum, and Alteromonadaceae in ${\gamma}$-Proteobacteria subphylum. Among eight strains of Bacillus spp., three strains showed more than 97% identity, but five strains showed about 90% identity with type strain on the basis of partial 16S rDNA sequence.