• Title/Summary/Keyword: l6S rDNA

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Phylogeny of Korean Rhus spp. Based on ITS and rbcL Sequences (ITS 및 rbcL 염기서열에 근거한 한국 자생 옻나무속의 계통분류)

  • Lee, Won-Kyung;Kim, Myong-Jo;Heo, Kweon
    • Korean Journal of Medicinal Crop Science
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    • v.12 no.1
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    • pp.60-66
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    • 2004
  • This study was carried out to confirm the phylogenetic relationships in Korean Rhus species. Sequences from internal transcribed spacers (ITS) of nuclear ribosomal DNA and rbcL gene of chloroplast DNA were determined. Cotinus coggygria was selected as outgroup because it is closest allied with Rhus in Anacardiaceae. Also, ingroup was limited as six Korean Rhus species. ITS 1 sequences in six species of Rhus and one species of Cotinus ranged from 246 to 253 bp and ITS 2 sequences from 234 to 244 bp. Concerning the G+C content of the studied taxa, ITS 1 sequences ranged from 58.0 to 68.13% and ITS 2 from 59.75 to 68.46%. On the other hand, rbcL sequences were same size in the all species examined by 1,428 bp. G+C contents of rbcL sequences were ranged from 43.56 to 43.77% which means there are nearly no different from interspecies each other. Phylogenetic tree strongly supports the colse relationships between R. succedanea and R. sylvestris. Rhus javanica and Cotinus coggygria were also closely allied with each other in ITS and rbcL trees. Therefore, R. javanica was regarded as most primitive species among the Korean Rhus species. ITS 1 region of nuclear ribosomal DNA was suggested as very useful taxonomical marker for genus Rhus.

Molecular Identification of Vaginal Lactobacillus spp. Isolated from Korean Women

  • CHANG, CHUNG EUN;SYLVIA I. PAVLOVA;LIN TAO;EUN-KI KIM;SEUNG CHUL KIM;HYUN SHIK YUN;JAE-SEONG SO
    • Journal of Microbiology and Biotechnology
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    • v.12 no.2
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    • pp.312-317
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    • 2002
  • Indigenous lactobacilli were isolated from vaginas of Korean women for possible use in ecological treatment of bacterial vaginosis. Vaginal swab samples were obtained from a gynecological clinic and streaked on Rogosa SL agar plates to select the most predominant lactobacilli in each sample. The preliminary identification of the isolates as lactobacilli was based on microscopic observation of Gram-positive rod-shaped cell morphology. The initial characterization was performed on 108 isolates in terms of their cell surface hydrophobicity (CSH), antimicrobial activity, and hydrogen peroxide (H₂O₂) production capability, and 10 isolates were then selected for further molecular identification. For a rapid procedure to identify lactobacilli, polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analyses of the l6S rRNA genes were applied. The 10 selected lactobacilli and 9 different reference strains of Lactobacillus spp. were characterized by PCR-RFLP where the amplified l6S rDNA was digested with 7 different restriction endonucleases prior to analysis. DNA sequencing of the 16S rRNA gene of one particular isolate, KLB 46, that had been identified as L. crispatus by the PCR-RFLP analysis, further confirmed its identity as L. crispatus.

A Study on the Freshwater Algal Flora Occurring in Temporary Ponds around the Dasan Arctic Station, Ny-Alesund (Norway), and the Molecular Characteristics of Chlamydomonas 18S rDNA (노르웨이 북극다산기지 주변에 형성된 일시적 담수지의 미세조류 및 Chlamydomonm 18S rDNA의 유전자 특성)

  • Ki, Jang-Seu;Kang, Sung-Ho;Jung, Sung-Won;Park, Bum-Soo;Han, Myung-Soo
    • Ocean and Polar Research
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    • v.28 no.2
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    • pp.107-117
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    • 2006
  • Freshwater algal studies in North polar environments are relatively few. This study presented the algal-flora, -biomass and genetic features of dominant cells collected from temporary ponds around the Polar Research Station (PRS), Norway. Water samples were collected from 4 stations around PRS, and analyzed for their environmental and biological variables. Water temperature, salinity and conductivity ranged from 5 to $10^{\circ}C$, 0.1 to $0.3%_{\circ}$ and 0.21 to $0.36{\mu}S/cm$, respectively. Chlorophyll a concentration ranged from 1.8 to $11.1{\mu}g/l$, and that of the size-fractionated cells was recorded from 0.7 to $1.1{\mu}g/l$ in picoplankton 0.3 to $6.5{\mu}g/l$ in nanoplankton, and 0.4 to $3.9{\mu}g/l$ in microplankton respectively. Algal flora in the present study was recorded as 10 genera, in which Chlamydomonas, particularly, was dominant in all studied sites. By comparison of Chlamydomonas 18S rDNA sequences, including two isolates from PRS, they formed a distinct clade against others: sequence similarity was significantly low (<97.2%) with C. noctigama, being the highest score by BLAST search in GenBank. This study was valuable for basic knowledge regarding the freshwater algae around PRS and their genetic information.

Analysis of Endospore-forming Bacteria or Nitrogen-fixing Bacteria Community Isolated from Plants Rhizosphere in Dokdo Island (독도서식 식물근권에서 분리한 포자형성세균과 질소고정세균의 군집구조 분석)

  • Jeon, Seon-Ae;Sung, Hye-Ri;Park, Yu-Mi;Pak, Jae-Hong;Ghim, Sa-Youl
    • Microbiology and Biotechnology Letters
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    • v.37 no.3
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    • pp.189-196
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    • 2009
  • Bacteria were isolated from roots of plants belonging to family Solanaceae and Gramineae, inhabited in Dokdo island. Fifty six endospore-forming bacteria grown on tryptic soy broth (TSB) agar medium and 23 nitrogen-fixing bacteria (NFB) grown on nitrogen free agar medium were isolated, respectively. The isolates were partially identified by analyzing the 16S rDNA and categorized into phylogenetic groups. The 16S rDNA sequences of each identified isolates were compared with sequences of each type strains to analyze phylogenetic relationship by phylogenetic tree. As a result, endospore-forming bacteria and nitrogen-fixing bacteria were classified into 4 and 6 lineage groups, respectively. Among these isolated, 18 were presumed to be novel species candidates based on the similarity (lower than 98%) analysis of the l6S rDNA sequences.

Detection and genetic characterization of Lawsonia intracellularis from swine in Korea

  • Chu, Jia-Qi;Hu, Xu-Min;Kim, Myung-Cheol;Park, Chang-Sik;Jun, Moo-Hyung
    • Korean Journal of Veterinary Service
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    • v.33 no.3
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    • pp.223-231
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    • 2010
  • A total of 191 samples collected from the commercial swine farms located in Chungnam province were investigated by PCR to estimate the prevalence of Lawsonia (L.) intracellularis infection. In the group of the pigs with proliferative enteritis, 14 (93.3%) of 15 intestinal samples and 12 (80.0%) of 15 feces were positive in PCR. In contrast, a relatively low positive rate (18.0%, 29 of 161 samples) was determined in the group of normal healthy pigs. The group of pigs over 120 days showed the highest positive rates (26.8%, 15 of 56 samples). In the comparison of the sequences of 210bp for species specific fragments and 301bp for outer membrane protein, the isolates (L1. L2) showed almost 100% identity with the reference L. intracellularis (L08049, USA). For the sequences of partial 16s rDNA, the homologies among the 5 isolates (L1-L5) were 97.4% to 99.3%, and those of 5 sequences (L1-L5) versus 5 overseas reference strains of L. intracellularis ranged from 98.6% to 99.8%. In the comparison of the nucleotide sequences among 5 isolates and other species in Desulfovibrionales showed 82.4 to 99.5% identities. The 5 isolates shared relatively low identities (76.9% to 84.4%) with the species of alpha-proteobacteria. In phylogenetic analysis based on the 16s rDNA sequences, all of the 5 isolates (L1-L5) were located in the same branch with the strains of L. intracellularis that were previously isolated from the pigs in USA and China. Seven strains of Desulfovibrio sp. were clustered in the neighboring branches, whereas alpha and gamma Proteobacteria showed distant relationship with L. intracellularis strains. The present findings suggest that L. intracellularis infection is endemic in the swine farms in the regions, and that the domestic isolates maintained very limited genetic variation.

Cytogenetic Mapping of Carthamus tinctorius L. with Tandemly Repeated DNA Sequences by Fluorescence in situ Hybridization

  • Mancia, Franklin Hinosa;Ju, Yoon Ha;Lim, Ki-Byung;Kim, Jung Sun;Nam, Sang Yong;Hwang, Yoon-Jung
    • Korean Journal of Plant Resources
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    • v.30 no.6
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    • pp.654-661
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    • 2017
  • Dual-color fluorescence in situ hybridization karyotype analysis was created using repetitive sequences including two types of rDNA repeats (45S and 5S rDNAs) and Arabidopsis-type telomere sequence repeats. The somatic metaphase cells of Carthamus tinctorius were observed as diploids (2n=2x=24). A symmetrical or slightly asymmetrical karyotype with seven pairs of metacentric and five pairs of submetacentric chromosomes was observed. The lengths of the somatic metaphase chromosomes ranged from 4.18 to $6.53{\mu}m$, with a total length of $60.71{\mu}m$. One locus of 45S rDNA was located on the pericentromeric regions of three pairs of chromosomes and the other pair was situated on the terminal regions of the short arms of a single pair of chromosomes. One locus of 5S rDNA was detected on the interstitial regions of the short arms of two pairs of chromosomes. Arabidopsis-type telomeric repeats were detected on the terminal regions of all pairs of chromosomes. Co-localization of loci between telomeric repeats and 45S rDNA was observed in a single pair of chromosomes. The results provide additional information for the existing physical mapping project of C. tinctorius and will also serve as a benchmark to a more intricate cytogenetic investigation of C. tinctorius and its related species.

A Method for Comparing Multiple Bacterial Community Structures from 16S rDNA Clone Library Sequences

  • Hur, Inae;Chun, Jongsik
    • Journal of Microbiology
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    • v.42 no.1
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    • pp.9-13
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    • 2004
  • Culture-independent approaches, based on 16S rDNA sequences, are extensively used in modern microbial ecology. Sequencing of the clone library generated from environmental DNA has advantages over fingerprint-based methods, such as denaturing gradient gel electrophoresis, as it provides precise identification and quantification of the phylotypes present in samples. However, to date, no method exists for comparing multiple bacterial community structures using clone library sequences. In this study, an automated method to achieve this has been developed, by applying pair wise alignment, hierarchical clustering and principle component analysis. The method has been demonstrated to be successful in comparing samples from various environments. The program, named CommCluster, was written in JAVA, and is now freely available, at http://chunlab.snu.ac.kr/commcluster/.

Identification and Characterization of Alginate Lyase Producing Pseudomonas sp. N7151-6 (Alginate Lyase 생산 균주 Pseudomonas sp. N7151-6의 분리 및 특성)

  • Lee, Jae-Hyung;Bae, Min-Ji;Kim, Yang-Choon;Nam, Soo-Wan
    • Microbiology and Biotechnology Letters
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    • v.37 no.4
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    • pp.350-354
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    • 2009
  • A Gram-negative, alginate lyase-producing bacterium was isolated from the Haeundae Coast, Korea. The isolated strain N7151-6 produced alginate lyase. The optimal temperature and pH for growth were found to be $30^{\circ}C$ and pH 8.0, respectively. This strain can be grown at the NaCl concentration of 0-7% (w/v). Analysis of 16S rDNA sequence and physiological profiling indicated that the strain N7151-6 belonged to Pseudomonas sp. The enzyme alginate lyase produced by Pseudomonas sp. N7151-6 was partially purified by ultrafiltration (MWCO= 30 kDa). The optimum pH and temperature for the activity of the purified enzyme were found to be 7.0 and $30^{\circ}C$, respectively. The enzyme was stable at the pH range of 5.0-9.0 and temperature range of $23-30^{\circ}C$. The total activity of alginate lyase produced was reached about 110 unit/L.

Karyotype Analyses of a Rice Cultivar 'Nakdong' and its Four Genetically Modified Events by Conventional Staining and Fluorescence in situ Hybridization

  • Jeon, Eun Jin;Ryu, Kwang Bok;Kim, Hyun Hee
    • Korean Journal of Breeding Science
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    • v.43 no.4
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    • pp.252-259
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    • 2011
  • Conventional staining and fluorescence in situ hybridization (FISH) karyotypes of the non-genetically modified (GM) parental rice line, 'Nakdong' (Oryza sativa L. japonica), and its four GM rice lines, LS28 (event LS30-32-20-1), Cry1Ac1 (event C7-1-9-1), and LS28 ${\times}$ Cry1Ac1 (events L/C1-1-3-1 and L/C1-3-1-1) were analyzed using 5S and 45S rDNAs as probes. Both parental and transgenic lines were diploids (2n=24) with one satellite chromosome pair. The lengths of the prometaphase chromosomes ranged from 1.50 to $6.30{\mu}m$. Four submetacentric and eight metacentric pairs comprised the karyotype of 'Nakdong' and its four GM lines. One pair of 5S rDNA signals was detected near the centromeric region of chromosome g in both the parental and transgenic lines. The 45S rDNA signals were detected on the secondary constrictions of the satellite chromosome pair in both the parental and transgenic lines. There was no significant difference in chromosome size, length, and composition between 'Nakdong' and its four GM lines. This research was conducted as a preliminary study for chromosomal detection of transgenes in GM rice lines and would be useful for their breeding programs.

Annual Population Variation and Identification of Antibiotic-Resistant Bacteria in the Lower Lake Geumgang (금강호의 항생제 내성세균의 분포 및 동정)

  • Bae, Myoung-Sook;Choi, Gang-Guk;Park, Suhk-Hwan;Choi, Moon-Sul;Lee, Geon-Hyoung
    • The Korean Journal of Ecology
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    • v.27 no.5
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    • pp.283-289
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    • 2004
  • This study was conducted to evaluate the annual population variation and identification of antibiotic-resistant bacteria in the lower artificial Lake Geumgang from January to December, 2002. Samples were taken from the surface waters at 3 stations near the estuarine barrage. The results were as follows; the population densities of heterotrophic bacteria varied from 4.1±1.0×10² to 6.7±1.1×10³ cfu ml/sup -1/ during the investigation periods. The population densities of antibiotic-resistant bacteria ranged from 1.5±0.7×10 to 4.3±0.3×10³ cfu ml/sup -1/ for ampicillin; from 0 to 6.4±0.4× 10² cfu ml/sup -1/ for chloramphenicol; from 0 to 2.8±0.3×10³ cfu ml/sup -1/ for gentamicin; from 0 to 4.5±1.0×10³ cfu ml/sup -1/ for kanamycin; and from 1.0±0.4 × 10 to 2.3±0.5×10³ cfu ml/sup -1/ for streptomycin, respectively. Of the sixty isolates, 90% were Gram negative. Dominant genera by 16S rDNA analysis were identified Aeromonas spp. (14 strains), Bacillus spp. (6 strains), Enterobacter spp. (4 strains), and Stenotrophomonas spp. (6 strains). These strains were clustered into 12 groups based on relatedness by average linkage method. Of the 60 isolates, 85% had the resistance to ampicilin and 32% were shown resistance to more than 2 kinds of antibiotics.