• Title/Summary/Keyword: environmental DNA

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Analysis of the Genome of Symbiobacterium toebii by Pulsed-Field Gel Electrophoresis

  • Hong, Seung-Pyo;Park, Jong-Hoon;Kim, Yong-Seung;Hwang, Hae-Jun;Rhee, Sung-Keun;Lee, Seung-Goo;Sung, Moon-Hee;Esaki, Nobuyoshi
    • Journal of Microbiology and Biotechnology
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    • v.10 no.3
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    • pp.405-409
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    • 2000
  • We have studied the genome of an obligately commensal thermophile, Symbiobacterium toebii. The chromosome was extracted from pure cultures of S. toebii recently established. Total DNA of S. toebii was resolved by pulsed-field gel electrophoresis (PFGE) into discrete numbers of fragments by digenstion with the endonuclease SspI, SpeI, XbaI, and HpaI. Estimated sizes of fragments produced by the four enzymes and their sum consistently yielded a total genome size of 2.8 Mb. Because restriction endonucleases NotI and SwaI, recognizing 8 bp, released too many fragments, these enzymes could not be used for the estimation of the genome size. Considering no mobility of undigested genome under PFGE, the genome of S. toebii appears to be circular. The presence of extrachromosomal DNA in S. toebii was excluded by the results of the conventional 1% agarose gel electrophoresis and the field inversion gel electrophoresis of undigested S. toebii DNA.

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Morphological and genetic characterization and the nationwide distribution of the phototrophic dinoflagellate Scrippsiella lachrymosa in the Korean waters

  • Lee, Sung Yeon;Jeong, Hae Jin;You, Ji Hyun;Kim, So Jin
    • ALGAE
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    • v.33 no.1
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    • pp.21-35
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    • 2018
  • The phototrophic dinoflagellate genus Scrippsiella is known to have a worldwide distribution. Here, we report for the first time, the occurrence of Scrippsiella lachrymosa in Korean waters. Unlike the other stains of S. lachrymosa whose cultures had been established from cysts in the sediments, the clonal culture of the Korean strain of S. lachrymosa was established from motile cells. When the sulcal plates of S. lachrymosa, which have not been fully described to date, were carefully examined using scanning electron microscopy, the Korean strain of S. lachrymosa clearly exhibited the anterior sulcal plate (s.a.), right sulcal plate (s.d.), left sulcal plate (s.s.), median sulcal plate (s.m.), and posterior sulcal plate (s.p.). When properly aligned, the large subunit (LSU) rDNA sequence of the Korean strain of S. lachrymosa was ca. 1% different from those of two Norwegian strains of S. lachrymosa, the only strains for which LSU sequences have been reported. The internal transcribed spacer (ITS) rDNA sequence of the Korean strain of S. lachrymosa was also ca. 1% different from those of the Scottish and Chinese strains and 3% different from those of the Canadian, German, Greek, and Portuguese strains. Thus, the Korean S. lachrymosa strain has unique LSU and ITS sequences. The abundances of S. lachrymosa in the waters of 28 stations, located in the East, West, and South Sea of Korea, were quantified in four seasons from January 2016 to October 2017, using quantitative real-time polymerase chain reaction method and newly designed specific primer-probe sets. Its abundances were >$0.1cells\;mL^{-1}$ at eight stations in January and March 2016 and March 2017, and its highest abundance in Korean waters was $26cells\;mL^{-1}$. Thus, S. lachrymosa has a nationwide distribution in Korean waters as motile cells.

Sirt1 Promotes DNA Damage Repair and Cellular Survival

  • Song, Seung-Hyun;Lee, Mi-Ok;Lee, Ji-Seon;Oh, Je-Sok;Cho, Sung-Uk;Cha, Hyuk-Jin
    • Biomolecules & Therapeutics
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    • v.19 no.3
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    • pp.282-287
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    • 2011
  • Sirt1, a nicotinamide adenine dinucleotide ($NAD^+$)-dependent histone deacetylase, is known to deacetylate a number of proteins that are involved in various cellular pathways such as the stress response, apoptosis and cell growth. Modulation of the stress response by Sirtuin 1 (Sirt1) is achieved by the deacetylation of key proteins in a cellular pathway, and leads to a delay in the onset of cancer or aging. In particular, Sirt1 is known to play an important role in maintaining genomic stability, which may be strongly associated with a protective effect during tumorigenesis and during the onset of aging. In these studies, Sirt1 was generated in stably expressing cells and during the stimulation of DNA damage to examine whether it promotes survival. Sirt1 expressing cells facilitated the repair of DNA damage induced by either ionizing radiation (IR) or bleomycin (BLM) treatment. Fastened damaged DNA repair in Sirt1 expressing cells corresponded to prompt activation of Chk2 and ${\gamma}$-H2AX foci formation and promoted survival. Inhibition of Sirt1 enzymatic activity by a chemical inhibitor, nicotinamide (NIC), delayed DNA damage repair, indicating that promoted DNA damage repair by Sirt1 functions to induce survival when DNA damage occurs.

Application of DNA Content and Total Protein Concentration to Predict Blooms Caused by Cochlodinium polykrikoides (Dinophyceae) in Korean Coastal Waters (total DNA 및 단백질 함량변화에 의한 C. polykrikoides 조기적조 예측 응용)

  • Cho, Eun-Seob;Park, Yong-Kyu
    • Journal of Life Science
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    • v.14 no.2
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    • pp.255-262
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    • 2004
  • We applied nuclear DNA content stained with 4'-6'-diamidino-2-phenylindole (DAPI) and total protein concentration to predict the existence of Cochlodinium polykrikoides before huge blooms occurred, based on a short-term survey at sites in the South Sea. Fluctuations in environmental conditions and nutrients (nitrate, nitrite, and phosphate) were of a similar range, regardless of sampling sites or early and middle field observations. However, C. polykrikoides abundance was significantly different depending on the station, with a higher cell density of 34, 62, and 57 cells L$^{-1}$ at Stn C2, C5, and C6, respectively than what was found in early August, 2000. In mid August, 2000, the highest cell density of 547 cells L$^{-1}$ at Stn C3 was observed. The relationship between C. polykrikoides abundance, DAPI-stained DNA content, and total protein concentration was a positive correlation coefficient, in particular a higher positive correlation was exposed to even a smaller abundance of C. polykrikoides. These results suggest that DNA stained by DAPI and total protein concentration could play an important index in easily predicting the presence of C. polykrikoides before blooms.

Using Reverse Dot Hybridization Method and 16S rRNA Gene (16S rDNA) for Identifying the Food Poisoning Microorganism in Foods (Reverse dot hybridization 방법과 16S rRNA gene(16S rDNA)을 이용한 식품에서 식중독균의 탐색)

  • Kim, Min-Seong;Shin, Kyu-Chul;Lee, Hyung-Gu;Han, Myung-Soo;Min, Byung-Re;Choi, Yong-Keel
    • Korean Journal of Food Science and Technology
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    • v.35 no.3
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    • pp.470-474
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    • 2003
  • DNA sequence information on small-subunit rRNA gene (16S rDNA) obtained from food-poisoning bacterial culture was used to investigate the presence of bacterial pathogens in food. By reverse dot blot detection method, presence of food-poisoning bacteria could be confirmed on hybridization of digoxigenin-labeled 16S rDNA Polymerase Chain Reaction (PCR) primer product and biotin-labeled specific oligonucleotide probe. Escherichia coli, Bacillus cereus. and Salmonella sp. were used as the representative food-poisoning bacterial microorganisms. An oligonucleotide probe, based on the variable region of 16S rRNA gene, was used as the specific probe. These tools may be more useful than classic biochemical method for rapid identification of contaminated food.

Effects of Cadmium on the Gene Expression Profile in the Rat Basal Ganglia (카드뮴이 흰쥐 뇌기저핵의 유전자 발현에 미치는 영향)

  • Lee, Chae-Kwan
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.20 no.1
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    • pp.29-40
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    • 2010
  • This study was aimed at investigating the gene expression profile in basal ganglia of cadmium exposed rat based on cDNA array analysis. For cDNA array analysis, adult Sprague-Dawley male rats (350 ${\pm}$ 25 g) were intraperitoneally injected with 2.0 mg/kg body weight/day of CdCl2 (0.3 ml) for 5 days. For doserelated gene expression analysis rats were intraperitoneally injected with 0.0, 0.1, 0.3, 1.0 mg/kg body weight/day of CdCl$_2$ for 5 days. Control rats were injected with equal volume of saline. Cadmium concentration of brain was analyzed by atomic absorption spectrophotometer. For cDNA array, RNA samples were extracted from basal ganglia and reverse-transcribed in the presence of [${\alpha}$32P]-dATP. Membrane sets of the Atlas Rat 1.2 array II and Toxicology array 1.2 (Clontech, Palo Alto, CA) were hybridized with cDNA probe sets. RT-PCR was employed to validate the relative gene expression patterns obtained from the cDNA array. Northern blot hybridization methods were employed to assess the dose-related gene expression. Among the 2352 cDNAs, 671 genes were detected in both array sets and 63 genes of 38 classes showed significant (more than two fold) changes in expression. Thirty five of these genes were up-regulated and twenty eight were down-regulated in the cadmium exposed group. According to the dose-related gene expression analysis, heat shock 27 kDa protein (HSP27), neurodegeneration-associated protein 1 (Neurodap 1) genes were significantly up-regulated and melatonin receptor 1a (Mel1a), Kinesin family member 3C (KIF3C), novel kinesinrelated protein (KIF1D) genes were significantly downregulated even in the low-dose of cadmium exposed group (0.1 mg/kg body weight/day). Conclusions Sixty three genes detected in this study can give some more useful informations about the cadmium-induced neurotoxicity in the basal ganglia. As well as, HSP27, Neurodap1, Mel1a, KIF3C and KIF1D genes may be useful for the study of the cadmium-induced neurotoxicity because these genes showed dramatic changes of mRNA levels in response to the low dose of cadmium exposure.

Korea Barcode of Life Database System (KBOL)

  • Kim, Sung-Min;Kim, Chang-Bae;Min, Gi-Sik;Suh, Young-Bae;Bhak, Jong;Woo, Tae-Ha;Koo, Hye-Young;Choi, Jun-Kil;Shin, Mann-Kyoon;Jung, Jong-Woo;Song, Kyo-Hong;Ree, Han-Il;Hwang, Ui-Wook;Park, Yung-Chul;Eo, Hae-Seok;Kim, Joo-Pil;Yoon, Seong-Myeong;Rho, Hyun-Soo;Kim, Sa-Heung;Lee, Hang;Min, Mi-Sook
    • Animal cells and systems
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    • v.16 no.1
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    • pp.11-19
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    • 2012
  • A major concern regarding the collection and storage of biodiversity information is the inefficiency of conventional taxonomic approaches in dealing with a large number of species. This inefficiency has increased the demand for automated, rapid, and reliable molecular identification systems and large-scale biological databases. DNA-based taxonomic approaches are now arguably a necessity in biodiversity studies. In particular, DNA barcoding using short DNA sequences provides an effective molecular tool for species identification. We constructed a large-scale database system that holds a collection of 5531 barcode sequences from 2429 Korean species. The Korea Barcode of Life database (KBOL, http://koreabarcode.org) is a web-based database system that is used for compiling a high volume of DNA barcode data and identifying unknown biological specimens. With the KBOL system, users can not only link DNA barcodes and biological information but can also undertake conservation activities, including environmental management, monitoring, and detecting significant organisms.

Archaeogenetic Research of Excavated Human Bones from the Ancient Tombs (분묘 유적지 출토 인골에 대한 고고유전학 연구)

  • Jee, Sang Hyun;Chung, Yong Jae;Seo, Min Seok
    • Korean Journal of Heritage: History & Science
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    • v.41 no.1
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    • pp.99-108
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    • 2008
  • The paleogenetic analysis has become an increasingly important subject of archaeological, anthropological, biological as well as public interest. Recently, scientific research for human skeletal remains was more activated because of increasing awareness of the valuable archaeological information by the ancient DNA analysis. State of preservation of organic remains vary in different soil and burying environmental condition. Almost all available tissue disappear to analysis ancient DNA of bone in acidic soil caused by climate and geological features in Korea. Many preserved human remains excavated in the 'Heogwakmyo'(limelayered tomb of Chosun Dynasty Period) is able to explain through the relationship between burial conditions and bone survival form the burial method and ceremony. Ancient DNA analysis of excavated human bone form ancient tomb requires to remove contaminants such as microorganism's DNA and soil components that affect authentic results. Particularly, contamination control of contemporary human DNA is major serious problem and should verified by criteria of authenticity. In order to understand migration and culture of ancient population, when possible, ancient DNA studies needs to go abreast both radiocarbon and stable isotope studies because the dietary inferences will suggest ancient subsistence and settlement patterns. Also when the paleogenetic research supported with the arts and humanities research such as physical anthropology and archaeology, more valuable ancient genetic information is providing a unique results about evolutionary and population genetics studies to reconstruct the past.

Purification, Characterization, and cDNA Cloning of Xylanase from Fungus Trichoderma Strain SY

  • Min, Shin-Young;Kim, Bong-Gyu;Lee, Chan;Hur, Hor-Gil;Ahn, Joong-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.12 no.6
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    • pp.890-894
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    • 2002
  • A xylanase-producing Trichoderma strain was isolated from soil. Xylanase from Trichoderma strain SY was purified 21-fold to an apparent homogeneity, with a $17.4\%$ yield. The optimum pH and temperature were determined to be 5.5 and $50^{\circ}C$, respectively, and its molecular weight was 21-kDa by SDS-PAGE. The corresponding gene, named xyl, was cloned by RT-PCR. DNA blot analysis of xyl showed that this gene is present as a single copy. The amino acid sequence of the Xyl protein showed similarity to those of other xylanases derived from various fungi. mRNA of xyl was highly expressed when this fungus was grown on cellulose or xylan as a sole carbon source, but undetectable when grown on sucrose. Extracts of Escherichia coli cells expressing xyl were found to have xylanase activity. It was confirmed that xyl from this isolate encodes xylanase.