• Title/Summary/Keyword: DNA migration

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Detection of DNA Rearrangement in Rice Using a Cosmid Library

  • Mun, Eun-Pyo;Nahm, Baek-Hie
    • Animal cells and systems
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    • v.1 no.4
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    • pp.629-635
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    • 1997
  • Previously we reported the migration and rearrangement of a chloroplast gene cluster into mitochondria. The exact genomic locations of the clusters, modes of the gene rearrangement and mechanisms of the interorganellar migration of the clusters have yet to be understood. The detailed analysis needs to include a larger region of DNA surrounding each cluster. To study DNA rearrangement and migration in more detail a cosmid library was constructed using the total rice genomic DNA including nuclear, chloroplast and mitochondrial DNA. From this cosmid library, a sub-library was obtained by selecting the clones hybridized to various regions of chloroplast DNA. According to the hybridization pattern 136 clones from the sub-library were classified into 29 groups. Detailed analysis of these clones revealed that in addition to authentic chloroplast DNA, the clones contain its homologs resulted from rearrangement and mutation. We analyzed two clones in detail, which contain different rp12 homologs resulted from rearrangement and/or migration, respectively.

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Mitochondrial DNA polymorphism in Fourteen Geographical Strains of Drosophila melanogoater (세계 14지역 계통에 대한 초파리 미토콘드리아 DNA의 다형현상)

  • 김봉기
    • The Korean Journal of Zoology
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    • v.31 no.3
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    • pp.218-224
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    • 1988
  • Restriction endonucleases were used to search for intraspecific variation at 32 cleavage sites in mitochondrial DNA(mtDNA) purified from fourteen strains of Drosophila melanogaster helonging to different localities of the world. mtDNA of D. melanogaster was displayed site variation(Hpall, Haelll and Seal endonucleases) and length variation(maxirnum 550bp). Six genotypes, Ml, M2, M3, M4, M6 and M7, could be distinguished based on ihe site types witti a low average of intraspecific substitution rate (1.88%),but M5 type of Ogasawara strain in Japan was not detected in this study. A possible explanation for the low divergence was that mtDNA variation of fourteen strains in D. melanogaster could not he accumulated sufficiently owing to recent divergence of few individuals, and that sequence divergence was prevented by frequent migration in spite of the geographical isolation.

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Single Cell Gel Electrophoresis (comet assay) to Detect DNA Damage and Apoptosis in Cell Level (DNA damage와 Apoptosis를 정량화하는 단세포전기영동법)

  • 류재천;김현주;서영록;김경란
    • Environmental Mutagens and Carcinogens
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    • v.17 no.2
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    • pp.71-77
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    • 1997
  • The single cell gel electrophoressis(SCGE) assay, also known as the comet assay, is a rapid, simple, visual and sensitive technique for measuring and analysing DNA breakage in mammalian cells. The SCGE or comet assay is a promising test for the detection of DNA damage and repair in individnal cells. It has widespread potential applications in DNA damage and repair studies, genotoxicity testing and biomonitoring. In this microgel electrophoresis technique, cells are embedded in agarose gel on microscope slides, iysed and electrophoresed under alkaline conditions. Cells with increased DNA damage display increased migration of DNA from the nucleus towards the anode. The length of DNA migration indicates the amount of DNA breakage in the cell. The comet assay is also capable of identifying apoptotic cells which contain highly fragmented DNA. Here we review the development of the SCGE assay, existing protocols for the detection and analysis of comets, the relevant underlying principles determining the behaviour of DNA and the potential applications of the technique.

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Biogeography and Distribution Pattern of a Korean Wood-eating Cockroach Species, Cryptocercus kyebangensis, Based on Genetic Network Analysis and DNA Sequence Information

  • Park, Yung-Chul;Choe, Jae-Chun
    • Journal of Ecology and Environment
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    • v.30 no.4
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    • pp.331-340
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    • 2007
  • We examined the evolutionary and ecological processes shaping current geographical distributions of a Korean wood-eating cockroach species, Cryptocercus kyebangensis. Our research aims were to understand evolutionary pattern of DNA sequences, to construct genetic network of Cryptocercus kyebangensis local populations and to understand evolutionary and ecological processes shaping their current geographical distribution patterns via DNA sequence information and genetic networks, using sequence data of two genes (ITS-2 and AT region) from local populations of C. kyebangensis. The results suggest that the ITS-2 and AT region are appropriate molecular markers for elucidating C. kyebangensis geographic patterns at the population level. The MSN-A based on the ITS-2 showed two possible routes, the Hwaak-san and Myeongji-san route and the Seorak-san and Gyebang-san route, for migration of ancestral C. kyebangensis into South Korea. The MSNs (MSN-A and -B) elucidate migration routes well within South Korea, especially the route of Group I and Group II.

Transfer of Genetic Substance Through the Cell Wall of Geranium (Pelargonium zonale hybrids, 'Pinto Scarlet') Callus (제라니움 세포벽을 통한 유전물질의 전이)

  • 유장걸;소인섭;홍경애
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.1
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    • pp.29-34
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    • 1994
  • The possibility that DNA could move out of the single cells isolated from geranium (Pelargonium zonale hybrids, 'PintoScarlet') callus was determined by the elechophoretic DNA analysis after treatment of low pH, various concentrations of KNO$_3$, 2,4-D, and GA$_3$ followed by the centrifugal force, all of which are hewn to and the physico-chemical properties of the cell wall. The centrifugal force of l,800 xg was need for DNA migration after the above treatment, but 7k300 xg was required without the treatments. In this experiment the optimum concentration (300 mg/L) of sodium dodecyl sulfate (SDS) used as an anion detergent to collect the negatively charged DNA was very critical not to damage the cell wall It can be concluded that the centrifugal force played a key role for the DNA migration through the cell wall, and the treatments of low pH (4.0), 0.5% KNO$_3$, 1.5 mg/L GA$_3$and 1mg/L 2,4-D further increased the DNA migration.

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Capillary Electrophoresis of Single-stranded DNA

  • Choi, Hyun-Ju;Kim, Yong-Seong
    • Bulletin of the Korean Chemical Society
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    • v.24 no.7
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    • pp.943-947
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    • 2003
  • We have studied the migration behavior of single-stranded DNA using capillary gel electrophoresis under various conditions. It was found that optimum electric fields should be less than 150 V/cm for the good tradeoff between the separation time and the resolution. It seems that the gel matrix with the combination of different polymer average molecular weights is important to extend the maximum readable DNA bases. The total gel concentration less than 3.1% in the mixed gel system showed good separation efficiency up to 600 bases. The best result was obtained with the poy(ethylene)oxide (PEO) gel concentration of 1.2% of Mr 8,000,000 and 1.8% of Mr 600,000. We observed that the capillary length between 50 cm to 100 cm (effective length) should be employed for the optimization between the total DNA migration time and the maximum readable length. A trizma base-boric acid-ethlyenediaminetetraacetic acid (EDTA) (TBE) buffer was commonly used for DNA sequencing, but we found that 3-[tris(hydroxymethyl)methyl amino]-1-propane sulfonic acid (TAPS) buffer worked as well for the single-stranded DNA separation. Especially, TAPS buffer showed a good resolution for very short DNA bases (1 to 30 bases).

Effects of Abeliophyllum distichum Nakai Flower Extracts on Antioxidative Activities and Inhibition of DNA Damage

  • Ahn, Joungjwa;Park, Jae Ho
    • Korean Journal of Plant Resources
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    • v.26 no.3
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    • pp.355-361
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    • 2013
  • The present study attempts to evaluate antioxidant activities of extracts from Abeliophyllum distichum. Nakai flower. The samples were collected in Jangyyeon-myeon, Goesan-gun, Korea and extracted with either hot-water or ethyl acetate (EtOAC). In DPPH, hydroxyl radical scavenging activity and $Fe^{2+}$ chelating activity of EtOAC extracts were 93.41%, 98.43%, and 7.38%, while those of hot-water extracts were 86.93%, 41.33% and 47.68% at 200 ${\mu}g/ml$, respectively. In ${\varphi}X$-174 RF I plasmid DNA cleavage assay, the protective effects of EtOAC and hot-water extracts against oxidative DNA damage were 82% and 17% at 200 ${\mu}g/ml$, respectively. Both extracts showed the protective effect of DNA migration by oxidative stress in intracellular DNA migration assay. Both extracts had no cytotoxity in NIH3T3 cells. Several polyphenolic compounds were identified such as 2-methoxy-benzoic acid, vanillic acid, phytol and pulegone by GC/MS. These results indicated that extracts of Abeliophyllum distichum Nakai flower showed antioxidant activities and protective activities against oxidative DNA damage and showed the possibility to be used as an effective natural antioxidants.

Effect of Salinity-stratified Waters on Upward Migration and Ratio of Extracted DNA/RNA in Cochlodinium polykrikoides Margalef Based on the Ratio of Absorbance at 260 and 280nm (염분 구배가 Cochlodinium polykrikoides Margalef의 수직이동 및 DNA/RNA 비율에 미치는 영향)

  • Cho Eun Seob;Lee Young Sik
    • Journal of Life Science
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    • v.15 no.3 s.70
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    • pp.468-473
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    • 2005
  • The coastal regions of Yeosu, the South Sea of Korea, has occurred annually the red tide which is caused by potentially ichthyotoxic dinoflagellate C. polykrikoides, with a wide avenue for exchange with oceanic waters and freshwater runoff from Sumjin river. We attempted to examine the variability in response to vertical migration and concentration of extracted DNA/RNA of C. polykrikoides exposed to salinity-stratified waters. The experimental aquarium of the 60 liter was employed to culture C. polykrikoides. One aquarium was supplied with only sea water, the other was consisted of sea water and freshwater. Experiment was conducted for 5 days. In experimental column (mixture of freshwater and sea water), salinity was maintained to 20 at upper and approximately 30 at bottom during the period of this study. The fluctuation with related to dissolved oxygen and pH was similar pattern to both columns. Chlorophyll a was significantly higher value at upper than bottom. During 24h, chlorphyll a on experimental column was extremely high on the top as soon as lighting, compared with control. With elapsed time, the gap between experimental and control columns was a little. In darkness, chlorophyll a was not significantly different between upper and bottom, most cells appeared to randomly distribute on column regardless of water layer. Fluctuation with related to concentration of extracted DNA and RNA based on ratio of absorbance of 260 and 280 nm in experimental column was higher at final day or diel migration than control. These results implied that a large volume of freshwater could be associated with influence of concentration of DNA and RNA, in particular, rapid upward movement caused massive fish kills as soon as sunset.

Cloning, characterization, and expression of the macrophage migration inhibitory factor gene from the pacific abalone (Haliotis discus hannai) (북방전복 (Haliotis discus hannai) 의 대식세포이동저해인자 (MIF, Macrophage migration inhibitory factor) 유전자 동정 및 발현분석)

  • Park, Eun Hee;Shin, Eun-Ha;Kim, Young-Ok;Kim, Dong-Gyun;Kong, HeeJeong;Kim, Woo-Jin;An, Chul Min;Nam, Bo-Hye
    • The Korean Journal of Malacology
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    • v.32 no.4
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    • pp.241-247
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    • 2016
  • Macrophage Migration Inhibitory Factor (MIF) are well-defined role as unique cytokine and critical mediator in acute and chronic inflammatory diseases, autoimmune diseases. In this study, we isolated and characterized a full-length of MIF cDNA from the abalone (Haliotis discus hannai). The full-length cDNA of abMIF was of 1264 bp, consisting of a 5'-terminal UTR of 143 bp, an open reading frame of 360 bp and a 3-terminal UTR of 761 bp. The abalone MIF cDNA encodes a 119-amino acid polypeptide with a calculated molecular mass of 13.4 kDa and isoelectric point of 9.07. Multiple alignments and phylogenetic analysis with the deduced abalone MIF protein and showed strong homology with disk abalone (Haliotis discusdiscus). The deduced amino acid sequence of abMIF exhibited homology with other reported MIFs, such as 80%, with that of other disk abalone H. discus discus MIF gene. Quantitative real-time PCR (qRT-PCR) analysis indicated that abMIF was highly expression observed in hapatopacreas, intestine, foot, and gonad of normal conditioned abalone. Even though AbMIF mRNA level in hemocytes was low under the normal condition, it was sharply up-regulated and reached the maximum at 6 h post-infection with Vibrio parahaemolyticus, and then decreased at 24 h post-infection. This result indicates that abMIF plays an important role in responding in the innate immune system.

DNA Separation Chips Using Asymmetrically-Switched Nonuniform Electric Fields (비대칭 교차전기장의 불균일 분포를 이용한 DNA 분리 소자)

  • Yi, So-Yeon;Cho, Young-Ho
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.33 no.3
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    • pp.265-268
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    • 2009
  • We present the experimental study to realize a DNA separation chip using asymmetrically-switched nonuniform electric fields. The DNA separation chip redistributes DNA molecules within a specific area based on the size- and field-dependent nonlinearity of DNA drift velocity. The present chip is composed of a width variable channel to distribute nonuniform electric field, a DNA loading slit and a pair of electrodes to apply electric field. We focus on the design of DNA separation chips with identifying the nonlinearity of DNA drift velocity using three different DNA molecules (11.1kbp, 15.6kbp, and 48.5kbp) in the chips. It is demonstrated that different size of DNA shows different net migration in different direction under the asymmetrically-switched nonuniform electric field.