• 제목/요약/키워드: Environmental DNA

검색결과 1,798건 처리시간 0.034초

MCF-7 세포주의 γ선에 의한 DNA 손상 반응 유전자 발현 양상의 분석 (A DNA-Damage Response Gene Expression Analysis in MCF-7 followed by γ-Radiation)

  • 박지윤;황창일;박웅양;김진규;채영규
    • 환경생물
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    • 제23권1호
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    • pp.21-26
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    • 2005
  • Cell response to genotoxic agents is complex and involves the participation of different classes of genes including cell cycle control, DNA repair and apoptosis. In this report, we presented a approach to characterize the cellular functions associated with the altered transcript profiles of MCF-7 exposed to low-dose in vitro gamma-irradiation. We used the method of human 2.4 k cDNA microarrays containing apoptosis, cell cycle, chromatin, repair, stress and chromosome genes to analyze the differential gene expression characterization that were displayed by radiation-exposed cell, human breast carcinoma MCF-7 cell line, such as 4 Gy 4 hr, 8 Gy 4 hr, and 8 Gy 12 hr. Among these genes, 66 were up-regulated and 49 were down-regulated. Specific genes were concomitantly induced in the results. Cyclin dependent kinase 4 (Cdk4) is induced for starting the cell cycle. This regulation is required for a DNA damage­induced G1 arrest. In addition to, an apoptotic pathways gene Bcl-w was concomitantly induced. Mismatch repair protein homologue-l (hMLH1), a necessary component of DNA mismatch protein repair (MMR), in G2-M cell cycle checkpoint arrest. The present study provides new information on the molecular mechanism underlying the cell response to genotoxic stress, with relevance to basic and clinical research.

Cloning and Expression of Alkaline Phosphatase Gene from Schizosaccharomyces pombe

  • Kang, Sung-Won;Cho, Young-Wook;Park, Eun-Hee;Ahn, Ki-Sup;Lim, Chang-Jin
    • BMB Reports
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    • 제34권3호
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    • pp.262-267
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    • 2001
  • A cDNA coding alkaline phosphatase (AP) homologue was isolated from a cDNA library of Schizosaccharomyces pombe by colony hybridization. The nucleotide sequence of the cloned cDNA appeared to lack the N-terminal coding region. The genomic DNA encoding alkaline phosphatase homologue was isolated from S. pombe chromosomal DNA using PCR. The amplified DNA fragment was ligated into plasmid pRS315 to generate the recombinant plasmid pSW20. The DNA insert was subcloned as two smaller fragments for nucleotide sequencing. The sequence contains 2,789 by and encodes a protein of 532 amino acids with a molecular mass of 58,666 daltons. The S. pombe cells containing plasmid pSW20 showed much higher AP activity compared with the yeast cells with vector only This indicates that the cloned AP gene apparently encodes AP The predicted amino acid sequence of the S. pombe AP shares homology with those of other known APs.

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An integrated DNA barcode assay microdevice for rapid, highly sensitive and multiplex pathogen detection at the single-cell level

  • Jung, Jae Hwan;Cho, Min Kyung;Chung, So Yi;Seo, Tae Seok
    • 한국진공학회:학술대회논문집
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    • 한국진공학회 2013년도 제45회 하계 정기학술대회 초록집
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    • pp.276-276
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    • 2013
  • Here we report an integrated microdevice consisting of an efficient passive mixer, a magnetic separation chamber, and a capillary electrophoretic microchannel in which DNA barcode assay, target pathogen separation, and barcode DNA capillary electrophoretic analysis were performed sequentially within 30 min for multiplex pathogen detection at the single-cell level. The intestine-shaped serpentine 3D micromixer provides a high mixing rate to generate magnetic particle-pathogenic bacteria-DNA barcode labelled AuNP complexes quantitatively. After magnetic separation and purification of those complexes, the barcode DNA strands were released and analyzed by the microfluidic capillary electrophoresis within 5 min. The size of the barcode DNA strand was controlled depending on the target bacteria (Staphylococcus aureus, Escherichia coli O157:H7, and Salmonella typhimurium), and the different elution time of the barcode DNA peak in the electropherogram allows us to recognize the target pathogen with ease in the monoplex as well as in the multiplex analysis. In addition, the quantity of the DNA barcode strand (~104) per AuNP is enough to be observed in the laser-induced confocal fluorescence detector, thereby making single-cell analysis possible. This novel integrated microdevice enables us to perform rapid, sensitive, and multiplex pathogen detection with sample-in-answer-out capability to be applied for biosafety testing, environmental screening, and clinical trials.

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고구마 배양세포에서 Peroxiredoxin cDNA의 분리 및 발현 특성 (Molecular Cloning and Characterization of a Peroxiredoxin cDNA from Cell Cultures of Sweetpotato)

  • 박수영;류선화;권석윤;김종국;곽상수
    • Journal of Plant Biotechnology
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    • 제30권2호
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    • pp.135-141
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    • 2003
  • Peroxiredoxin(Pix) are large family of peroxidases that reduce alkyl hydroperoxides and hydrogen peroxide. A cDNA clone (referred to as swPrxl) encoding Pix was from a sweetpotato cDNA library constructed from suspension-sultured cells, and its expression was investigated in terms of stress. The swPrxl contained an open reading frame (ORF) encoding mature protein of 193 amino acids with calculated molecular mass of 20.8kDa. The predicted amino acid sequence of swPrxl has two conserved cysteines that are essential resicues for the reduction of peroxides. It showed high amino acid sequence homology ot PixIIF of Arabidopsis (77%) and putative Prx of rice(72%). RNA gel-blot analysis showed that swPrxl gene was expressed dominantly in leave among intact tissues, and also highly detect in suspension-cultured cells. Interestingly, the level of swPrxl transcripts was almost the same regardless of the growth stage in suspension culture. Furthermore, the transcription level of swPrxl gene was not significantly changed in response to various stress treatments such as wounding, extreme temperature and stress-related chemicals RT-PCR analyses.

토양 및 수계환경에서 Transformation에 의한 세균들간의 수평적 유전물질 전이 (Horizontal Gene Transfer among Bacteria by Transformation in Soil and Aquatic Environments)

  • 이건형
    • 환경생물
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    • 제18권2호
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    • pp.205-213
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    • 2000
  • 실험실에서 형질전환될 수 있는 세균들은 자연환경 조건에서도 형질전환 능력이 발달하는 것으로 알려져 있다. 따라서, 환경 내에서 형질전환 능력이 있는 세균의 존재는 확실한 것으로 여겨진다. DNA는 무기물에 부착된 상태에서는 핵산분해효소에 의한 분해로부터 보호되는 것으로 알려져 있다. 비록 DNA가 토양 속에 분산되어져 일정 비율로 가수분해되더라도, 수 주일 후에도 낮은 비율로 감지될 수 있다. 따라서 free DNA는 자연적 형질전환을 할 수 있을 만큼 충분히 지속될 수 있다. 실험실 조건에서는 세균의 형질전환이 여러 경우 보고되었지만, 자연상태에서 형질전환과 관련된 자료는 매우 적다. 생태학적으로 GMMs로부터 재조합 DNA가 토착 미생물에 전이될 수 있는 잠재력에 대한 문제가 주요 현안이 되었는데, 이는 전이된 DNA가 방출된 세균의 생태학적인 적응력을 변화시켜 생물학적 안전성의 문제를 야기할 수 있기 때문이다. 물론, 방출된 GMMs로부터 재조합 DNA가 토착 미생물에 전이되는 율은 아주 낮은 빈도로 일어나지만, 빈도가 낮다는 것은 그리 중요하지 않다. 왜냐하면, 비록 낮은 빈도로 전이되더라도 유리한 조건을 만나게 되면 전이된 유전자는 선택될 수 있기 때문이다. 이제까지 GMMs는 실험실이나 제한된 환경에서 주로 사용되었지만 앞으로는 개방된 자연 생태계에서 이루어질 전망이다. 그러므로 GMMs가 토착세균에 미치는 영향에 대해서도 연구되어야 하고 동시에 GMMs가 생태계에 방출될 경우 그에 따른 영향평가를 반드시 수행해야 한다.

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나노튜브전극을 사용한 전압전류법에 의한 식물잎에서 살충제 검출 (Detection of Pesticide Thiram in Plant Leafs Using Voltammetric at Nanotube Electrode)

  • 이장현;이수영
    • 한국환경과학회지
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    • 제19권12호
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    • pp.1335-1341
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    • 2010
  • Voltammetric diagnostics of pesticide thiram was studied in plant leafs in vivo fluid with DNA immobilized on a carbon nanotube electrode (DCE). Sensor properties of carbon nanotube (CE) and DNA immobilized nanotube were compared. DCE was more effective than CE in target detecting. The parameters such as pH strength, stripping accumulation, amplitude, and increment potential were examined to find the optimum condition for detection of pesticide thiram in a sesame leaf. The optimized conditions were as follows 550 Hz frequency, 0.15 V amplitude, 0.005 V increment potential, -1.2 V initial potential, 4.78 pH, 500 sec accumulation time. Under optimum condition, the detection limit of thiram was attained at 0.01ng/L.

비스페놀 A 및 Diethylstilbestrol의 유전독성 평가를 위한 염색체이상, 자매염색분체교환, 소핵형성, 단일세포 겔 전기영동법의 활용 (Analysis of chromosome aberration, sister chromatid exchange, micronuclei and single cell gel electrophoresis in human lymphocytes exposed in vitro to Bisphenol A and Diethylstilbestrol)

  • 김병모;정해원
    • 한국환경성돌연변이발암원학회지
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    • 제21권2호
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    • pp.135-141
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    • 2001
  • Endocrine disruptors have been implicated in carcinogenesis in animal studies, but carcinogenetic effects on human remain controversial. In order to examine the genotoxicity of two common endocrine disruptors, Bisphenol A and Diethylstilbestrol, cytogenetic endpoints including chromosome aberration (CA), sister chromatid exchange (SCE), micronuclei (MN) analyses and DNA damage by single cell gel electrophoresis (SCGE) were assessed. The effects of Bisphenol A and Diethylstilbestrol on the frequencies of CA and MN were increased in a dose-dependent manner and that of Bispheol A was more significant by Kendall'$\tau$test. Bisphenol A and Diethylstilbestrol also increased the frequency of SCE. Bisphenol A and Diethylstilbestrol induced DNA damage in a dose-dependent manner and the DNA damage induced by Diethylstilbestrol in human blood lymphocytes was more significant.

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적응반응 관련 DNA 회복유전자의 발현조절에 관한 연구 (Regulatory Expression of DNA Repair Genes Involved in Adaptive Response)

  • 최수영;이희원;박상대
    • 한국환경성돌연변이발암원학회지
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    • 제10권1호
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    • pp.1-10
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    • 1990
  • The regulation of DNA repair genes expression was investigated using fused genes, in which the promoter of repair genes was hybridized with the lacZ structural gene. The activities of beta-galactosidase expressed from the fused gense were highly increased when the host cells were exposed to methylating agents, such as methyl methansulfonate (MMS), N-methyl-N'-nitro-nitrosoguanidine (MNNG) and methyl nitrosourea (MNU). On the other hand, the enzyme activities from the fused genes were not induced when the cells were treated with ethylating or nonalkylating agents, such as ethyl methansulfonate (EMS), 4-nitroquinoline-1-oxide (4NQO), Bleomycin, and Benzo(a)pyrene (BP).

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니켈의 독성과 발암성 (Nickel Toxicity and Carcinogenicity)

  • 박형숙;박광식
    • Environmental Analysis Health and Toxicology
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    • 제19권2호
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    • pp.119-134
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    • 2004
  • Human exposure to highly nickel-polluted environments, such as those associated with nickel refining, electroplating, and welding, has the potential to produce a variety of pathologic effects. Among them are skin allergies, lung fibrosis, and cancer of the respiratory tract. The exact mechanisms of nickel-induced carcinogenesis are not known and have been the subject of numerous epidemiologic and experimental investigations. This review provides the evidence of the current state for the genotoxic and mutagenic activity of Ni (II) particularly at high doses. Such doses are best delivered into the cells by phagocytosis of sparingly soluble nickel-containing dust particles. Ni (II) genotoxicity may be aggravated through the generation of DNA-damaging reactive oxygen species (ROS) and the inhibition of DNA repair by this metal. The epigenetic effects of nickel includes alteration in gene expression resulting from DNA hypermethylation and histone hypoacetylation, as well as activation some signaling pathways and subsequent transcrziption factors.