• Title/Summary/Keyword: Repetitive DNA

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Genetic and Phenotypic Diversity of Dichlorprop-Degrading Bacteria Isolated from Soils

  • Park, Hae-Dong;Ka, Jong-Ok
    • Journal of Microbiology
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    • v.41 no.1
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    • pp.7-15
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    • 2003
  • Nine dichlorprop-degrading bacteria and three pairs of bacteria showing syntrophic metabolism of the herbicide were isolated from soils, and their genetic and phenotypic characteristics were investigated. Analysis of 16S rDNA sequences indicated that the isolates were related to members of the genera, Sphingomonas, Herbaspirillum, and Bradyrhizobium. Twelve different chromosomal DNA patterns were obtained by polymerase-chain-reaction (PCR) amplification of repetitive extragenic palindromic (REP) sequences from the 15 isolates. The isolates were able to utilize the herbicide dichlorprop as a sole source of carbon and energy and their dichlorprop derogative pathways were induced by the presence of dichlorprop. Most of the isolates and syntrophic pairs were able to degrade both (R)- and (S)-dichlorprop, but strain DP522 exhibited enantioselective degradation of (S)-dichlorprop. The isolates degraded 2,4-dichlorophenoxyacetic acid, 2-methyl-4-chlorophenoxyacetic acid , and mecoprop, in addition to dichlorprop. Oxygen uptake experiments indicated that most of the isolates degraded dichlorprop through 2,4-dichlorophenol.

UNDERSTANDING OF EPIGENETICS AND DNA METHYLATION (후생유전학 (Epigenetics)과 DNA methylation의 이해)

  • Oh, Jung-Hwan;Kwon, Young-Dae;Yoon, Byung-Wook;Choi, Byung-Jun
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.30 no.3
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    • pp.302-309
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    • 2008
  • Epigenetic is usually referring to heritable traits that do not involve changes to the underlying DNA sequence. DNA methylation is known to serve as cellular memory. and is one of the most important mechanism of epigenetic. DNA methylation is a covalent modification in which the target molecules for methylation in mammalian DNA are cytosine bases in CpG dinucleotides. The 5' position of cytosine is methylated in a reaction catalyzed by DNA methyltransferases; DNMTl, DNMT3a, and DNMT3b. There are two different regions in the context of DNA methylation: CpG poor regions and CpG islands. The intergenic and the intronic region is considered to be CpG poor, and CpG islands are discrete CpG-rich regions which are often found in promoter regions. Normally, CpG poor regions are usually methylated whereas CpG islands are generally hypomethylated. DNA methylation is involved in various biological processes such as tissue-specific gene expression, genomic imprinting, and X chromosome inactivation. In general. cancer cells are characterized by global genomic hypomethylation and focal hypermethylation of CpG islands, which are generally unmethylated in normal cells. Gene silencing by CpG hypermethylation at the promotors of tumor suppressor genes is probably the most common mechanism of tumor suppressor inactivation in cancer.

Structure and Diversity of Arsenic-Resistant Bacteria in an Old Tin Mine Area of Thailand

  • Jareonmit, Pechrada;Sajjaphan, Kannika;Sadowsky, Michael J.
    • Journal of Microbiology and Biotechnology
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    • v.20 no.1
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    • pp.169-178
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    • 2010
  • The microbial community structure in Thailand soils contaminated with low and high levels of arsenic was determined by denaturing gradient gel electrophoresis. Band pattern analysis indicated that the bacterial community was not significantly different in the two soils. Phylogenetic analysis obtained by excising and sequencing six bands indicated that the soils were dominated by Arthrobacter koreensis and $\beta$-Proteobacteria. Two hundred and sixty-two bacterial isolates were obtained from arsenic-contaminated soils. The majority of the As-resistant isolates were Gramnegative bacteria. MIC studies indicated that all of the tested bacteria had greater resistance to arsenate than arsenite. Some strains were capable of growing in medium containing up to 1,500 mg/l arsenite and arsenate. Correlations analysis of resistance patterns of arsenite resistance indicated that the isolated bacteria could be categorized into 13 groups, with a maximum similarity value of 100%. All strains were also evaluated for resistance to eight antibiotics. The antibiotic resistance patterns divided the strains into 100 unique groups, indicating that the strains were very diverse. Isolates from each antibiotic resistance group were characterized in more detail by using the repetitive extragenic palindromic-PCR (rep-PCR) DNA fingerprinting technique with ERIC primers. The PCR products were analyzed by agarose gel electrophoresis. The genetic relatedness of 100 bacterial fingerprints, determined by using the Pearson product-moment similarity coefficient, showed that the isolates could be divided into four clusters, with similarity values ranging from 5-99%. Although many isolates were genetically diverse, others were clonal in nature. Additionally, the arsenic-resistant isolates were examined for the presence of arsenic resistance (ars) genes by using PCR, and 30% of the isolates were found to carry an arsenate reductase encoded by the arsC gene.

Oxidative Stress Induces Hypomethylation of LINE-1 and Hypermethylation of the RUNX3 Promoter in a Bladder Cancer Cell Line

  • Wongpaiboonwattana, Wikrom;Tosukhowong, Piyaratana;Dissayabutra, Thasinas;Mutirangura, Apiwat;Boonla, Chanchai
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.6
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    • pp.3773-3778
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    • 2013
  • Increased oxidative stress and changes in DNA methylation are frequently detected in bladder cancer patients. We previously demonstrated a relationship between increased oxidative stress and hypomethylation of the transposable long-interspersed nuclear element-1 (LINE-1). Promoter hypermethylation of a tumor suppressor gene, runt-related transcription factor 3 (RUNX3), may also be associated with bladder cancer genesis. In this study, we investigated changes of DNA methylation in LINE-1 and RUNX3 promoter in a bladder cancer cell (UM-UC-3) under oxidative stress conditions, stimulated by challenge with $H_2O_2$ for 72 h. Cells were pretreated with an antioxidant, tocopheryl acetate for 1 h to attenuate oxidative stress. Methylation levels of LINE-1 and RUNX3 promoter were measured by combined bisulfite restriction analysis PCR and methylation-specific PCR, respectively. Levels of LINE-1 methylation were significantly decreased in $H_2O_2$-treated cells, and reestablished after pretreated with tocopheryl acetate. Methylation of RUNX3 promoter was significantly increased in cells exposed to $H_2O_2$. In tocopheryl acetate pretreated cells, it was markedly decreased. In conclusion, hypomethylation of LINE-1 and hypermethylation of RUNX3 promoter in bladder cancer cell line was experimentally induced by reactive oxygen species (ROS). The present findings support the hypothesis that oxidative stress promotes urothelial cell carcinogenesis through modulation of DNA methylation. Our data also imply that mechanistic pathways of ROS-induced alteration of DNA methylation in a repetitive DNA element and a gene promoter might differ.

Centromere Repeat DNA Originated from Brassica rapa is Detected in the Centromere Region of Raphanus sativus Chromosomes

  • Hwang, Yoon-Jung;Yu, Hee-Ju;Mun, Jeong-Hwan;Bok, Kwang;Park, Beom-Seok;Lim, Ki-Byung
    • Horticultural Science & Technology
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    • v.30 no.6
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    • pp.751-756
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    • 2012
  • Fluorescence in situ hybridization (FISH) is a powerful tool for the detection of DNA sequences in the specific region of the chromosomes. As well as for the integrated physical mapping, FISH karyotype analysis has to be preceded. Karyotype of Raphanus sativus 'Wonkyo 10039' was analyzed by a dual-color FISH technique; using various repetitive DNA probes, including 5S rDNA, 45S rDNA, and centromere retrotransposon. The length of the somatic metaphase chromosome ranged from 1.35 to $2.06{\mu}m$ with a total length of $15.29{\mu}m$. The chromosome complements comprised of eight pairs of metacentrics and one pair of submetacentric. Bleached DAPI Band analysis revealed a heterochromatin region, covering 28.6% to 50.4% each chromosomes. 5S and 45S rDNA sequences were located on two and three pairs of chromosomes, respectively. The centromere retrotransposon of Brassica (CRB) is a major component in Brassica related species that has been maintained as a common centromere component. CRB signals were detected on the centromere and pericentromeric region of R. sativus 'Wonkyo 10039' and three basic Brassica species (B. rapa, B. nigra, and B. oleracea). These results will provide a valuable background for physical mapping and elucidation of the evolutionary relationship among the Brassica related species.

Effect of Body Mass Index on Global DNA Methylation in Healthy Korean Women

  • Na, Yeon Kyung;Hong, Hae Sook;Lee, Duk Hee;Lee, Won Kee;Kim, Dong Sun
    • Molecules and Cells
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    • v.37 no.6
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    • pp.467-472
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    • 2014
  • Obesity is known to be strongly associated with cardiovascular disease and cancer, the leading causes of mortality worldwide, and develops owing to interactions between genes and the environment. DNA methylation can act as a downstream effector of environmental signals, and analysis of this process therefore holds substantial promise for identifying mechanisms through which genetic and environmental factors jointly contribute to disease risk. Global DNA methylation of peripheral blood cells has recently been proposed as a potential biomarker for disease risk. Repetitive element DNA methylation has been shown to be associated with prominent obesity-related chronic diseases, but little is known about its relationship with weight status. In this study, we quantified the methylation of Alu elements in the peripheral blood DNA of 244 healthy women with a range of body mass indexes (BMIs) using pyrosequencing technology. Among the study participants, certain clinical laboratory parameters, including hemoglobin, serum glutamic oxaloacetic transaminase, serum glutamic- pyruvic transaminase, total cholesterol, and triglyceride levels were found to be strongly associated with BMI. Moreover, a U-shaped association between BMI and Alu methylation was observed, with the lowest methylation levels occurring at BMIs of between 23 and $30kg/m^2$. However, there was no significant association between Alu methylation and age, smoking status, or alcohol consumption. Overall, we identified a differential influence of BMI on global DNA methylation in healthy Korean women, indicating that BMI-related changes in Alu methylation might play a complex role in the etiology and pathogenesis of obesity. Further studies are required to elucidate the mechanisms underlying this relationship.

Random amplified polymorphic DNA analysis of bacterial pathogens using universal rice primers

  • Monoldorova, Sezim;Kim, Jinsol;Kim, Joon Hee;Jeon, Bo-Young
    • Korean Journal of Veterinary Service
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    • v.40 no.1
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    • pp.1-6
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    • 2017
  • Molecular typing of pathogenic microorganisms is important for epidemiological investigation of infectious disease outbreaks. In this study, we applied Universal Rice Primers (URP) that were originated from repetitive sequences in rice chromosomal DNA to random amplified polymorphic DNA (RAPD) analysis of pathogenic bacteria such as Escherichia coli, Listeria monocytogenes, and Salmonella sp. Of the twelve URP primers examined to date, seven primers (URP-2, -3, -4, -5, -6, -8, and -9) generated reproducible and polymorphic PCR products ranging from 1 to 13 bands. One of them, URP-6 was very effective in differentiating seven E. coli serotypes, seven L. monocytogenes clinical isolates, and eight Salmonella subspecies (ssp.) serovars. The results thus indicate that RAPD analysis using URP primers might be useful in typing bacterial pathogens including E. coli, L. monocytogenes, and Salmonella strains.

Genetic Divesity Analysis of Fungal Species by Universal Rice Primer (URP)-PCR (Universal Rice Primer (URP)-PCR에 의한 곰팡이 종의 유전적 다양성 검정)

  • Kang, Hee-Wan
    • The Korean Journal of Mycology
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    • v.40 no.2
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    • pp.78-85
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    • 2012
  • URP primers that were derived from repetitive DNA sequence of rice weedy rice have been applied for producing PCR polymorphisms in different fungal species. URP-PCR protocol employed stringent PCR with high annealing temperature over $55^{\circ}C$ throughout the thermo-cycling reaction, giving high reproducibility. Under the PCR condition, Each single URP primer produced characteristic fingerprints from diverse genomes of different fungal species, indicating its universal applicability. URP-PCR has been accessed for applicability to various fungal species with 33 genus, 142 species and 1,489 isolates. Numerous related papers have demonstrated that URP-PCR profiles of fungal species are very useful for identifying fungal species at intra and inter species levels. The results were reviewed in this paper.

Molecular and Cellular Studies of Seed Storage Proteins from Rice and Wheat

  • Kim, Woo-Taek
    • Applied Biological Chemistry
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    • v.32 no.1
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    • pp.64-72
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    • 1989
  • Near full length cDNA clones encoding the rice seed storage protein, prolamine, were isolated and divided into two homology classes based on cross-hybridization and DNA sequencing analysis. These cDNA clones contain a single open reading frame encoding a putative rice prolamine precursor(M.W.=17,200) possessing atypical 14 amino acid signal peptide. Clones of these two homology classes diverge mainly by insertions/deletions of short nucleotide stretches and point mutations. The deduced primary structures of both types of prolamine polypeptides are devoid of any major tandem repetitive sequences, a feature prevalent in other cereal prolamines. No significant homology teas detected between the rice prolamine and other cereal prolamines, indicating that the rice gene evolved from a different ancestor that gave rise to other cereal prolamine genes. Developing wheat and rice endosperms were examined using ultrathin sections prepared from tissues harvested at various days after flowering. By immunocytochemical localization techniques, wheat prolamines are localized within vesicles from Golgi apparatus and in homogeneous regions of protein bodies. The involvement of the goli apparatus in the packaging of wheat prolamines into protein bodies indicates a pathway which differs from the mode of other cereal prolamines and resembles the mechanism employed for the storage of rice glutelin and legume globulins.

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Isolation and Characterization of 4-(2,4-Dichlorophenoxy)Butyric Acid-Degrading Bacteria from Agricultural Soils

  • Park, In-Hyun;Ka, Jong-Ok
    • Journal of Microbiology and Biotechnology
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    • v.13 no.2
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    • pp.243-250
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    • 2003
  • Eight numerically dominant 4-(2,4-dichlorophenoxy) butyric acid (2,4-DB)-degrading bacteria and three pairs of bacteria showing syntrophic metabolism of 2,4-DB were isolated from soils, and their phylogenetic and phenotypic characteristics were investigated. The isolates were able to utilize 2,4-DB as a sole source of carbon and energy, and their 2.4-DB degradative enzymes were induced by the presence of 2.4-DB. Analysis of 16S rDNA sequences indicated that the isolates were related to members of the genera, Variovorax, Sphingomonas, Bradyrhizobium, and Pseudomonas. The chromosomal DNA patterns of the isolates obtained by polymerase-chain-reaction (PCR) amplification of repetitive extragenic palindromic (REP) sequences were distinct from each other. Four of the isolates had plasmids, but only one strain, DB 1, rad a transmissible 2,4-D degradative plasmid. When analyzed with PCR using primers targeted to the tfdA, B, and C genes, only strains DB2 and DB9a produced DNA bands of the expected sizes with the tfdA and C primers, respectively. All of the isolates were able to degrade 2,4-D as well as 2,4-DB, suggesting that the degradation pathways of these compounds were closely related to each other, but respiratory activities of many isolates adapted to 2,4-DB metabolism were quite low with 2,4-D.