• Title/Summary/Keyword: 환경 DNA

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Isolation of HRD3 gene, a homologous RAD3 gene from fission yeast Schizosaccharomyces pombe

  • Choi, In-Soon;Jin, Yong-Hwan;Park, Sang-Dai
    • Environmental Mutagens and Carcinogens
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    • v.16 no.2
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    • pp.77-82
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    • 1996
  • The RAD3 gene of Saccharomyces cerevisiae is required for excision repair and is essential for cell viability. RAD3 encoded protein possesses a single stranded DNA-dependent ATPase and DNA-RNA helicase activies. To examine the extent of conservation of structure and function of RAD3 during eukaryotic evolution, we have cloned the RAD3 homolog, HRD3, from the distantly related yeast Schizosaccharomyces pombe. Here, we report the partial cloning and characterization of HRD3 gene (Homologous of RAD3 gene) which was isolated by PCR amplification using conserved domain of Saccharomyces cerevisiae RAD3 gene. Chromosomal DNA isolated from S. pombe had similar restriction patterns to those from S. cerevisiae, as determined by Southern blot analysis. The 2. 8 kb transcript of mRNA was identified by Northern hybridization. The level of transcript did not increase upon UV-irradiation, suggesting that the HRD3 gene in S. pombe is not UV-inducible.

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Oxidative DNA damage by Ethanol Extract of Green Tea

  • Park You-Gyoung;Kwon Hoonjeong
    • Environmental Mutagens and Carcinogens
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    • v.25 no.2
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    • pp.71-75
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    • 2005
  • Green tea and their major constituents such as catechins are famous materials for their anti-oxidative and anti-carcinogenic activity, but many compounds with reducing power can promote the oxidation in their oxidized form or in the presence of metal ion. We investigated the pro-oxidative effect of the ethanol extract equivalent up to 30mg of dried weight of green tea leaves in four in vitro systems which could be used for detecting DNA damage. Although ethanol extract of green tea did not show significant mutagenicity in Salmonella typhimurium TA102, which is sensitive strain to oxidative stress, it degraded deoxyribose extensively in the presence of $FeCl_3-EDTA$ complex, promoted 8-oxoguanine formation in the live bacteria cell, Salmonella typhimurium TAI04, and cleaved super coiled DNA strand with the help of copper ion. It suggested that green tea, famous anti-oxidative material, can be pro-oxidant according to the condition of extraction or metal existence.

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Cytogenic Effects of Transplacentally Administered 2-Bromopropane -Pattern of Replicative DNA Synthesis(RDS) by BrdU Labeling Method- (2-Bromopropane의 경태반 영향에 관한 연구 -마우스 태자로의 이행과 태자세포의 복제 DNA합성세포에 관하여-)

  • 김영환;배은상
    • Journal of environmental and Sanitary engineering
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    • v.13 no.3
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    • pp.37-42
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    • 1998
  • 2-Bromopropane has been implicated to be the reason for the mass intoxication of workers at an electronic company in Korea. 2-Bromopropane deposition and pattern of DNA replication in mouse fetuses were analyzed after intravenous injection of 2-bromopropane. Injections were administered to pregnant ICR mice in order to cytogenetically evaluate transplacental 2-bromopropane. The results are summarized as follows; 1. A dose-dependent effect on DNA replication was observed equally in the lung, liver and kideneys of fetuses has been exposed to 2-bromopropane transplacentally as reductions of the labeling index. 2. Deposition of transplacentally administred 2-bromopropane in the fetus was lower than placenta.

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디클로벤지딘에 폭로된 흰쥐의 간장세포와 방광 상피세포에 형성된 DNA adducts의 $^{32}P-postlabeling$과 GC/MS-SIM에 의한 분석

  • 이진헌;신호상;장미선
    • Proceedings of the Korean Environmental Health Society Conference
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    • 2002.04a
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    • pp.49-51
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    • 2002
  • To identify and evaluate the dichlorobenzidine(DCB)-DNA adducts in liver cell and bladder epithelial cells by $^{32}$ P-postlabeling and GC/MS-SIM, we orally exposed the dichlorobenzidine (20mg/kh body wt.,/day)to male sprague-dawley rats for 14 days. Two kinds of DCB-DNA adduct were found at the same site of thin layer chromatogram of $^{32}$ P-postlabeling method in liver cells and bladder epithelial cells. In liver cells, relative adduct labeling(RAL) $\times$ 10$^{12}$ of DCB-DNA adduct A1 were 34.1$\pm$3.71 and 69.9$\pm$5.02, that of adduct A2 were 74.1$\pm$10.1 and 105.1$\pm$10.1 on 10 and 14 days after treatment, respectively. And in bladder epithelia cells, RAL $\times$ 10$^{12}$ of DCB-DNA adduct A1 were 5.92$\pm$1.60 and 15.9$\pm$1.31, that of adduct A2 were 9,81$\pm$2.81 and 22.8$\pm$1.79 on 10 and 14 days after treatment, respectively. DCB metabolites formed DNA adducts were monoacetyl-dichlorobenzidine(acDCB) and diacety1-dichlorobenzidine(di-acDCB), which was identify by gas chromatography/mass spectrometry-scan ionization mode(GC/MS-SIM), along with hydrolysis, extraction and TFA(trifluoroacetyl anhyride) derivatization with DCB-DNA adducts isolated from live cells and bladder epithelial cells. The base peak of acDCB were 252 and 294 m/z, and that of di-acDCB were 252, 294 and 336 m/z. In conclusion, the exposed DCB formed two kinds of DCB-DNA adduct, the proximate materials of that were acDCB and di-acDCB in liver and bladder epithlial cells. And the above GC/MS-SIM method was found the DCB-DNA adducts could be monitoring by gas chromatography.

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A Possible Target for the Heat Inactivation of SCK Tumor Cells

  • 강만식;정주영
    • The Korean Journal of Zoology
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    • v.32 no.4
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    • pp.305-313
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    • 1989
  • The present investigation aims at inquiring into a possible target for the heat inactivation of SCK tumor cells by comparing the kinetics of cell survival, rate of protein synthesis, and DNA polymerase activity in the presence of heat protector or heat sensitirer. A possible conclusion to be drawn from the present experiment is that there is no direct correlation between cell death and decrease in the rate of protein synthesis, but that the loss of DNA polvmerase $\beta$ activity correlates quite well with cell inactivation. Thus, protein degrada-tion and/or abnormal protein synthesis causes cell inactivation innireuv, possibly by altering the cellular environment which in turn affects the DNA polymerase $\beta$ activity. Accordingly, further studies, dealing with the correlation between changes in the cellular environment and DNA polymerase $\beta$ activity, are needed to set insight into a possible target for the heat inactivation of cells. 본 연구는 열보호제 또는 열증감제의 존재하에서 세포 생존곡선, 단백질 합성률, DNA 중합효소 $\beta$의 활성변화를 비교 검토함으로써 SCK 종양세포가 열에 의해서 불활성화될 때의 표적이 무엇인지를 밝혀보기 위해서 수행되었다. 본 실험의 결과로 추정하건대 열에 의한 세포치사는 단백질 합성률의 변화와는 직접적인 연관성이 없으나, DNA 중합효소 $\beta$의 활성도와는 밀접한 연관성이 있음을 알 수 있다. 즉, 단백질의 분해 또는 비정상적인 단백질의 합성이 세포의 환경을 변화시키고 이것이 DNA 중합효소 $\beta$의 활성에 영향을 미침으로써 간접적으로 세포의 치사를 초래할 것으로 짐작할 수 있다. 따라서, 세포의 열불화성화의 표적을 좀더 분명히 밝히기 위해서는 세포의 환경변화와 DNA 중합효소 $\beta$의 활성과의 관계를 추구하는 연구가 수행되어야 할 것으로 사료된다.

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Phylogenetic study of the Genus Suaeda(Chenopodiaceae) based on chloroplast and nuclear DNA sequences from Korea (엽록체 DNA 및 핵 DNA 염기서열에 근거한 한국산 나문재속(명아주과)의 분류학적 연구)

  • Kim, Suk-Kyu;Chung, Sang Ok
    • Korean Journal of Environment and Ecology
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    • v.32 no.6
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    • pp.566-574
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    • 2018
  • The purpose of this study was to identify the phylogenetic relationships of the plants in the Korean genus Suaeda and to find out the molecular markers that could confirm the interspecies relationships in the family tree through molecular phylogenetic studies. We used the nuclear ribosomal DNA ITS and the chloroplast DNA matK, psbA-trnH, and trnL-trnF as the molecular markers. We could not distinguish between S. japonica and S. maritima and between S. maritima and S. australis in the ITS region and could not distinguish between S. japonica and S. australis with the base sequence in the psbA-trnH and trnL-trnF region. However, we analyzed the combinations of four molecular marker regions and confirmed that each of five plant species of the genus Suaeda formed the independent line. Therefore, it is considered that combinations of molecular markers would be useful for the analysis of phylogenetic relationships in the genus Suaeda. Further investigations of the ecological and morphological characteristics would be needed to understand the phylogenetic relationship and lineage diversification in the genus Suaeda.

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.

Optimization of DNA Extraction and PCR Conditions for Fungal Metagenome Analysis of Atmospheric Particulate Matter (대기 입자상물질 시료의 곰팡이 메타게놈 분석을 위한 DNA 추출 및 PCR 조건 최적화)

  • Sookyung Kang;Kyung-Suk Cho
    • Microbiology and Biotechnology Letters
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    • v.51 no.1
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    • pp.99-108
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    • 2023
  • Several challenges arise in DNA extraction and gene amplification for airborne fungal metagenome analysis from a particulate matter (PM) samples. In this study, various conditions were tested to optimize the DNA extraction method from PM samples and polymerase chain reaction (PCR) conditions with primer set and annealing temperature. As a result of comparative evaluation of DNA extraction under various conditions, chemical cell lysis using buffer and proteinase K for 20 minutes and bead beating treatment were followed by using a commercial DNA extraction kit to efficiently extract DNA from the PM filter samples. To optimize the PCR conditions, PCR was performed using 10 primer sets for amplifying the ITS2 gene region. The concentration of the PCR amplicon was relatively high when the annealing temperature was 58℃ with the ITS3tagmix3/ITS4 primer set. Even under these conditions, when the concentration of the PCR product was low, nested PCR was performed using the primary PCR amplicon as the template DNA to amplify the ITS2 gene at a satisfactory concentration. Using the methods optimized in this study, DNA extraction and PCR were performed on 15 filter samples that collected PM2.5 in Seoul, and the ITS2 gene was successfully amplified in all samples. The optimized methods can be used for research on analyzing and interpreting the fungal metagenome of atmospheric PM samples.

Epigenetic Mechanisms of Depression: Role of Histone Modification and DNA Methylation in BDNF Gene (우울증의 후성유전기전: BDNF 유전자의 히스톤 변형 및 DNA 메틸화의 역할)

  • Park, Sung Woo
    • Journal of Life Science
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    • v.28 no.12
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    • pp.1536-1544
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    • 2018
  • Depression is a common, serious, and recurring mental disorder. The pathogenesis of depression involves many factors such as environmental factor, genetic factor and alteration of structure and function in neurobiological systems. Increasing evidence supports that epigenetic alteration may be associated with depression. The epigenetics is explained as the mechanisms by which environmental factor causes changes in chromatin structure and alters gene expression without changing DNA base sequence. DNA methylation and histone modification involving histone acetylation and methylation are the main epigenetic mechanisms. Animal studies have shown that stressful environment such as early life stress can leave persistent epigenetic marks in the genome, which alter gene expression and influence neural and behavioral function through adulthood. A potentially important gene in depression is brain-derived neurotrophic factor (BDNF). BDNF plays a central role in depression and antidepressant action. In studies of the rodent, exposure to stress at prenatal, postnatal, and adult stages alters BDNF expression through histone modification and DNA methylation of the BDNF gene which results in anxiety and depressive-like behavior. This review discusses recent advances in the study of the epigenetic mechanisms that contribute to depression, particularly histone modification and DNA methylation of the BDNF gene, that may help in the development of new targets for depression treatment.