• 제목/요약/키워드: induced hybridization

검색결과 181건 처리시간 0.02초

Heterologous Microarray Hybridization Used for Differential Gene Expression Profiling in Benzo[a]pyrene-exposed Marine Medaka

  • Woo, Seon-Ock;Won, Hyo-Kyoung;Jeon, Hye-Young;Kim, Bo-Ra;Lee, Taek-Kyun;Park, Hong-Seog;Yum, Seung-Shic
    • Molecular & Cellular Toxicology
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    • 제5권4호
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    • pp.283-290
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    • 2009
  • Differential gene expression profiling was performed in the hepatic tissue of marine medaka fish (Oryzias javanicus) after exposure to benzo[a]pyrene (BaP), a polycyclic aromatic hydrocarbon (PAH), by heterologous hybridization using a medaka cDNA microarray. Thirty-eight differentially expressed candidate genes, of which 23 were induced and 15 repressed (P<0.01), were identified and found to be associated with cell cycle, development, endocrine/reproduction, immune, metabolism, nucleic acid/protein binding, signal transduction, or non-categorized. The presumptive physiological changes induced by BaP exposure were identified after considering the biological function of each gene candidate. The results obtained in this study will allow future studies to assess the molecular mechanisms of BaP toxicity and the development of a systems biology approach to the stress biology of organic chemicals.

3중 염색체 probe를 이용한 FISH(fluorescence in situ hybridization)기법으로 분석한 정상인의 염색체 이상빈도 (Frequency of Chromosome Aberrations Detected by Fluorescence in Situ Hybridization Using Triple Chromosome-Specific Probes in o Healthy Korean Population)

  • 정해원;김수영;신은희
    • 한국환경성돌연변이발암원학회지
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    • 제18권2호
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    • pp.109-115
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    • 1998
  • Fluorscence in situ hybridization with chromosome-specific probe has been shown to be a valid and rapid method for detection of chromosome rearrangements induced by chemical and physical agents. This method is useful for quantifying structural aberrations, expecially for stable ones, such as translocation and insertion, which are difficult to detect with conventional method in human lymphocyte. In order to use the FISH method as a biodosimeter for monitoring human population exposed to various chemical and physical agent, baseline level of chromosome rearragement was established. Blood from forty four healthy adults were collected and analysed with whole chromosome-specific probes by human chromosome 1,2 and 4. The frequencies of stable translocation were 2.45 per 100 cell equivalent and those of insertion, color juction, acentric and dicentric were 0.32, 3.28, 0.23 and 0.27 per 100 cell equivalent respectively. The frequencies of chromosome rearragements increased with age in both sexes except for dicenrics. From above result, stable aberrations accumulate with age and it may reflect integrated lifetime exposure of adverse environment.

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Expression of EuNOD-ARP1 Encoding Auxin-repressed Protein Homolog Is Upregulated by Auxin and Localized to the Fixation Zone in Root Nodules of Elaeagnus umbellata

  • Kim, Ho Bang;Lee, Hyoungseok;Oh, Chang Jae;Lee, Nam Houn;An, Chung Sun
    • Molecules and Cells
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    • 제23권1호
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    • pp.115-121
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    • 2007
  • Root nodule formation is controlled by plant hormones such as auxin. Auxin-repressed protein (ARP) genes have been identified in various plant species but their functions are not clear. We have isolated a full-length cDNA clone (EuNOD-ARP1) showing high sequence homology to previously identified ARP genes from root nodules of Elaeagnus umbellata. Genomic Southern hybridization showed that there are at least four ARP-related genes in the genome of E. umbellata. The cDNA clone encodes a polypeptide of 120 amino acid residues with no signal peptide or organelle-targeting signals, indicating that it is a cytosolic protein. Its cytosolic location was confirmed using Arabidopsis protoplasts expressing a EuNOD-ARP1:smGFP fusion protein. Northern hybridization showed that EuNOD-ARP1 expression was higher in root nodules than in leaves or uninoculated roots. Unlike the ARP genes of strawberry and black locust, which are negatively regulated by exogenous auxin, EuNOD-ARP1 expression is induced by auxin in leaf tissue of E. umbellata. In situ hybridization revealed that EuNOD-ARP1 is mainly expressed in the fixation zone of root nodules.

Review on Functionalization of Laser-Induced Graphene

  • Jin Woo An;Hee Jae Kim;Seoung-Ki Lee
    • 한국전기전자재료학회논문지
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    • 제36권3호
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    • pp.203-213
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    • 2023
  • Owing to carbon materials' diverse functionalization and versatility, the design and synthesis of carbon-based three-dimensional porous structures have become important foundational research topics across various fields. Among the various methods for producing porous carbon structures, laser-induced graphene (LIG) has garnered attention because of its large surface area, controllable structure, excellent electrical conductivity, scalability, and eco-friendly synthesis process. In addition, recent research results have reported more novel functionalities by advancing further from the unique characteristics of LIG through functionalization or compounding of LIG, making it an attractive material for various applications in electronic devices, sensing, catalysis, and energy storage. This review aims to update the research trends in LIG and its functionalization, providing insights to inspire more interesting studies on functional LIG to expand its potential applications ultimately. Starting with the synthesis method and material characteristics of LIG, we introduce the functionalization of LIG, which is classified into surface modification, heteroatom doping, and hybridization based on the interaction mechanism. Finally, we summarize and discuss the prospects of LIG and its functionalization.

Identification of Genes that are Induced after Cadmium Exposure by Suppression Subtractive Hybridization

  • 이미옥
    • 한국환경독성학회:학술대회논문집
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    • 한국환경독성학회 2003년도 춘계학술대회
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    • pp.107-107
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    • 2003
  • The heavy metal cadmium is a xenobiotic toxicant of environmental and occupational concern and it has been classified as a human carcinogen. Inhalation of cadmium has been implicated in the development of emphysema and pulmonary fibrosis, but, the detailed mechanism by which cadmium induces adverse biological effects is not yet known. Therefore, we undertook the investigation of genes that are induced after cadmium exposure to illustrate the mechanism of cadmium toxicity For this purpose, we employed the polymerase chain reaction-based suppression subtractive hybridization technique. We identified 29 different cadmium-inducible genes in human peripheral mononuclear cells, such as macrophage migration inhibitory factor, lysophosphatidic acid acyltransferase-${\alpha}$, enolase-1${\alpha}$, VEGF, Bax, neuron-derived orphan receptor-1, and Nur77, which are known to be associated with inflammation, cell survival, and apoptosis. Induction of these genes by cadmium treatment was further confirmed by semi-quantitative reverse-transcription polymerase chain reaction. Further, we found that these genes were also induced after cadmium exposure in normal human lung fibroblast cell line, WI-38, suggesting potential use of this induction profile to monitor cadmium toxicity in the lung. Next, Nur77, one of cadmium-inducible genes, was further studied since the products of Nur77 are known to be involved in the apoptotic process of lung cells. Following cadmium treatment, Nur77 gene expression was increased at protein-level in A549 cells. Consistently, the reporter containing Nur77 binding sequence was activated by 2.5-fold after exposure to cadmium in reporter gene analysis by transient transfection experiments. When the plasmid encoding dominant negative Nur77 that represses the transcriptional function of wild-type Nur77 was transfected into A549 cells, the expression of Bax was significantly reduced, suggesting that induction of Nur77 was an important process in cadmium-induced apoptosis in the cells. Cadmium induced the expression of Nur77 in vivo, confirming the relevance of the data obtained in viro. Together our results suggest that Nur77 gene expression in exposure to cadmium leads apoptosis of lung cells which may cause pathological changes in lung.

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DNA-Functionalized Polymers and Nanoparticles for Gene Sensing

  • Maeda, Mizuo
    • 한국고분자학회:학술대회논문집
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    • 한국고분자학회 2006년도 IUPAC International Symposium on Advanced Polymers for Emerging Technologies
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    • pp.33-34
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    • 2006
  • The graft copolymer consisting of poly(N-isopropylacrylamide) (PNIPAAm) and single-stranded DNA was prepared. Interestingly, the copolymer was found to form nanoparticles above physiological temperature. We found that non-crosslinking aggregation of the nanoparticles was induced by the hybridization of the surface-bound DNA with the full-match complementary DNA, but not with one-base mismatch. The core material is not restricted to PNIPAAm; DNA-functionalized gold nanoparticle was found to show a similar aggregation induced only by the fully-complementary DNA, resulting in rapid color change within 3 min at ambient temperature. This methodology is general in principle and applicable for wide variety of clinical gene diagnosis.

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Schizosaccharomyces pombe에서 SNF2에 속하는 hrp2+ 유전자의 특성 연구 (Characterization of hrp2 + Gene Related to SNF2 Family in Schizosaccharomyces pombe)

  • Park, In-Soon
    • 한국환경성돌연변이발암원학회지
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    • 제22권3호
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    • pp.137-141
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    • 2002
  • 본 연구는 분열형 효모 Schizosaccharomyces pombe에서 여러 가지 DNA 절제회복 및 유전자 발현에 관여하는 SNF2/SW12유전자의 기능을 연구하기 위하여 이에 관련되는 유전자를 분리하고 그 특성을 연구하였다. SNF2 motif 의 conserved sequence를 primer로 하여 중합효소 연쇄반응(PCR) 방법으로 480 bp 크기의 DNA fragment를 분리하여, 이를 probe로 하여 효모에서 hrp2+ 유전자를 분리하였다. 분리한 hrp2+ 유전자의 sequence homology를 비교한 결과 3개의 SNF2 motif를 포함하고 있었다. hrp2+유전자의 전사체 크기는 4.7 kb임을 Northern hybridization으로 확인하였다. hrp2+유전자의 전사 개시 부위를 알기 위하여 primer extension분석을 한 결과, 첫 번째 ATG에서 약47 base pair 위쪽에 위치함을 확인하였다. 또한 특성 연구를 위하여 Northern hybridization으로 hrp2+ 유전자의 UV와 MMS에 대한 유도성을 조사한 결과 자외선에 대해서만 유전자의 발현이 유도되었다. 이 결과 분리한 hrp2+는 UV-inducible 유전자임을 확인하였다.

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Schizosaccharomyces pombe에서 SNF2에 속하는 hrp2+ 유전자의 특성 연구 (Isolation and Characterization of hrp2+ Gene Related to SNF2 Family In Yeast)

  • 최인순
    • 생명과학회지
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    • 제15권2호
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    • pp.192-196
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    • 2005
  • 본 연구는 분열형 효모 Schizosaccharomyces pombe에서 여러 가지 DNA 절제회복 및 유전자 발현에 관여하는 SNF2/SW12유전자의 기능을 연구하기 위하여 이에 관련되는 유전자를 분리하고 그 특성을 연구하였다. SNF2 motif의 conserved sequence를 primer로 하여 중합효소 연쇄반응 (PCR) 방법으로 480 bp 크기의 DNA fragment를 분리하여, 이를 probe로 하여 효모에서 hrp2+ 유전자를 분리하였다. 분리한 hrp+ 유전자의 sequence homology를 비교한 결과 3개의 SNF2 motif를 포함하고 있었다. hrp2+ 유전자의 전사체 크기는 4.7kb임을 Northern hybridization으로 확인하였다. 분리한 유전자의 특성 연구를 위하여 Northern hybridization 으로 hrp2+ 유전자의 UV와 MMS에 대한 유도성을 조사한 결과 자외선에 대해서만 유전자의 발현이 유도되었다. 이 결과 분리한 hrp2+는 UV-inducible 유전자임을 확인하였다. 또한 분리한 유전자의 특성연구 중 하나로 hrp2+ 단백질을 분리하여 helicase activity를 측정하였다. 이 결과 분리한 hrp2+ 유전자는 전혀 helicase activity를 나타내지 않았다.

Identification of Differentially Expressed Genes by Exposure of Methylmercury in Neuroblastoma Cell Line Using Suppression Subtractive Hybridization (SSH)

  • Kim, Youn-Jung;Ryu, Jae-Chun
    • Molecular & Cellular Toxicology
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    • 제2권1호
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    • pp.60-66
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    • 2006
  • Methylmercury (MeHg), one of the heavy metal compounds, can cause severe damage to the central nervous system in humans. Many reports have shown that MeHg is poisonous to human body through contaminated foods and has released into the environment. Despite many studies on the pathogenesis of MeHg-induced central neuropathy, no useful mechanism of toxicity has been established so far. This study, using of suppression subtractive hybridization (SSH) method, was peformed to identify differentially expressed genes by MeHg in SH-SY5Y human neuroblastoma cell line. We prepared to total RNA from SH-SY5Y cells treated with solvent (DMSO) and $6.25\;{\mu}M\;(IC_{50})$ MeHg and performed forward and reverse SSH. Differentially expressed cDNA clones were screened by dot blot, sequenced and confirmed that individual clones indeed represent differentially expressed genes with real time RT-PCR. These sequences were identified by BLAST homology search to known genes or expressed sequence tags (ESTs). Analysis of these sequences may provide an insight into the biological effects of MeHg in the pathogenesis of neurodegenerative disease and a possibility to develop more efficient and exact monitoring system of heavy metals as ubiquitous environmental pollutants.

Gene expression involved in dark-induced leaf senescence in zoysiagrass (Zoysia japonica)

  • Cheng, Xiaoxia;Dai, Xiaomei;Zeng, Huiming;Li, Yunxia;Tang, Wei;Han, Liebao
    • Plant Biotechnology Reports
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    • 제3권4호
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    • pp.285-292
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    • 2009
  • Zoysiagrass (Zoysia japonica) is one of the important turfgrass species. Extending green period of zoysiagrass via delaying leaf senescence will make this species have more potential in the turfgrass industry. In this study, we found that zoysiagrass seedlings treated with $GA_3$ could delay the leaf senescence induced by darkness. To study expression of genes responsive to staying green in zoysiagrass, suppression subtractive hybridization (SSH) was used to identify differentially expressed genes between non-$non-GA_3-treated$ and $GA_3-treated$ seedlings subjected to darkness. A total of 307 ESTs were generated, of which 226 ESTs clustered into 54 contigs and 81 were singlets. Differentially expressed genes selected by subtractions were classified into six categories according to their putative functions generated by BLAST analysis. Expression of five selected genes, Met, SAM, V-ATPase, Cry (Cryptochrome gene), and An (diphthine synthase gene) were examined by RT-PCR and Real-time PCR. Both RT-PCR and Real-time PCR results demonstrated that the differential expressions of these genes were attributable to delaying senescence by exogenously applied gibberellic acid. This is the first genome-wide study of senescence in a species of turfgrass.