• Title/Summary/Keyword: ProteinChip

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Developing a Protein-chip for Depigmenting Agents Screening (미백제 스크리닝용 단백질칩의 개발)

  • Kim, Eun-Ki;Kwak, Eun-Young;Han, Jung-Sun;Lee, Hyang-Bok;Shin, Jung-Hyun
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.31 no.1 s.49
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    • pp.13-16
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    • 2005
  • For the high-throughput-screening system (HTS) of depigmenting agents using a protein chip, effects of oligonucleotide-inhibitor sequence on the binding of Mitf protein to E box of MC1R was investigated. The sequence of oligonucletide-inhibitor affected the binding of the target DNA to Mitf, depending on the location of the sequence variation in the inhibitor nucleotide. The oligonucletide-inhibitor that changed the CATGTG sequence didn't show enough inhibition of the target DNA to Mitf, whereas significant inhibition was observed when the sequence outside the CATGTG was changed. This result indicated that CATCTG is crucial sequence for the binding of Mitf to I-box which initiates the transcription of pigmenting genes.

Interaction of Phenolic Compound-Specific Activator with Its Promoter using SPR-Based DNA Chip (SPR 근거 DNA 칩을 이용한 페놀 화합물 특이 CapR 조절 단백질과 촉진유전자와의 상호작용 연구)

  • 박선미;박후휘;임운기;신혜자
    • Journal of Life Science
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    • v.13 no.1
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    • pp.99-104
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    • 2003
  • Aromatic compounds are of major concern among environmental pollutants due to their toxicity and persistence. To monitor aromatic compounds in a real time with a better sensitivity, a new method of SPR (surface plasmon resonance) based on DNA chip (Biacore 3000) was developed here. It is thought that CapR regulatory protein as a complex with phenol, could bind to their corresponding promoter, Po. Biotinylated DNA oligomers for the promoter was synthesized by PCR and coupled onto streptoavidin-linked CM5-chip. CapR regulatory proteins were purified after cloning their genes in pET21a (+) vector and expressing proteins. The interaction was assessed by the system where the regulatory proteins flowed with or without phenol through the cells of DNA chip. CapR regulatory protein even in the presence of phenol had no response to its promoter, Po, suggesting that other factor(s) might be required for the activation of Po promoter. The present work reveals a promising possibility of the SPR-based DNA chip in monitoring specific environmental pollutants in a real time.

Development and evaluation of surface plasmon resonance imaging for the detection of antibodies against classical swine fever virus in swine

  • Cho, Ho-Seong;Lee, Tae-Uk;Park, Nam-Yong
    • Korean Journal of Veterinary Service
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    • v.30 no.2
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    • pp.205-209
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    • 2007
  • A protein chip based on surface plasmon resonance (SPR) imaging was developed for measuring classical swine fever virus (CSFV) antibody using a recombinant gp55 protein as an antigen. The diagnostic potential of SPR imaging for detecting antibodies to the CSFV gp55 protein was compared with that of a enzyme -linked immunosorbent assay (ELISA) using 70 pig sera. There was a strong positive correlation between the SPR imaging and ELISA (n=70, r=0.916, p<0.01). Therefore, the SPR imaging, which is a label-free and high-through put method, is expected to be a valuable tool in the serodiagnosis of CSFV.

Regulation of RIP3 protein stability by PELI1-mediated proteasome-dependent degradation

  • Park, Han-Hee;Morgan, Michael J.;Kang, Ho Chul;Kim, You-Sun
    • BMB Reports
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    • v.51 no.10
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    • pp.484-485
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    • 2018
  • Receptor-interacting protein kinase-3 (RIP3 or RIPK3) is a serine-threonine kinase largely essential for necroptotic cell death; it also plays a role in some inflammatory diseases. High levels of RIP3 are likely sufficient to activate necroptotic and inflammatory pathways downstream of RIP3 in the absence of an upstream stimulus. For example, we have previously detected high levels or RIP3 in the skin of Toxic Epidermal Necrolysis patients; this correlates with increased phosphorylation of MLKL found in these patients. We have long surmised that there are molecular mechanisms to prevent anomalous activity of the RIP3 protein, and so prevent undesirable cell death and inflammatory effects when inappropriately activated. Recent discovery that Carboxyl terminus of Hsp 70-Interacting Protein (CHIP) could mediate ubiquitylation- and lysosome-dependent RIP3 degradation provides a potential protein that has this capacity. However, while screening for RIP3-binding proteins, we discovered that pellino E3 ubiquitin protein ligase 1 (PELI1) also interacts directly with RIP3 protein; further investigation in this study revealed that PELI1 also targets RIP3 for proteasome-dependent degradation. Interestingly, unlike CHIP, which targets RIP3 more generally, PELI1 preferentially targets kinase active RIP3 that has been phosphorylated on T182, subsequently leading to RIP3 degradation.

Identification of Radiation-Sensitive Gene in U937 Cell by using cDNA-Chip Composed of Human Cancer Related Gene (U937 세포에서 발암관련 유전자들로 구성된 DNA chip을 이용한 방사선 감수성 유전자들의 선발)

  • 김종수;김인규;강경선;윤병수
    • Environmental Mutagens and Carcinogens
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    • v.22 no.1
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    • pp.54-59
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    • 2002
  • We have used cDNA microarray hybridization to identify gene regulated in response to gamma-irradiation in U-937 cell. The cDNA-chip was composed entirely of 1,000 human cancer related gene including apoptosis and angiogenesis etc. In gamma-irradiated U-937 cell, highly charged protein, ribosomal protein L32, four and a half LIM domains 3, lipocalin 2 (oncogene 24p3) and interleukin 15, ataxia telangiectasia mutated (includes complementation groups A, C and D) genes showed increased level of its transcription, and cell division cycle 25A, dihydrofolate reductase, topoisomerase (DNA) II beta(180kD), kinase suppressor of ras and strarigin genes showed reduced level of its transcription compared to untreated U-937 cell. The significant change of level of transcription was not found in well-known ionizing radiation(IR)-responsive gene, such as transcription factor TP53 and p53 related gene, except ataxia telangiectasia mutated gene.

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Development of Photo-sensor for Integrated Lab-On-a-Chip (집적화된 Lab-On-a Chip을 위한 광센서의 제작 및 특성 평가)

  • 김주환;신경식;김용국;김태송;김상식;주병권
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.17 no.4
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    • pp.404-409
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    • 2004
  • We fabricated photo-sensor for fluorescence detection in LOC. LOC is high throughput screening system. Our LOC screens biochemical reaction of protein using the immunoassay, and converts biochemical reaction into electrical signal using LIF(Laser Induced Fluorescence) detection method. Protein is labeled with rhodamine intercalating dye and finger PIN photodiode is used as photo-sensor We measured fluorescence emission of rhodamine dye and analyzed tendency of fluorescence detection, according to photo-sensor size, light intensity, and rhodamine concentration. Detection current was almost linearly proportional to two parameters, intensity and concentration, and was inversely proportional to photo-sensor size. Integrated LOC consists of optical-filter deposited photo-sensor and PDMS microchannel detected 50 (pg/${mu}ell$) rhodamine. For integrated LOC including light source, we used green LED as the light source and measured emitted fluorescence.

Feature Extraction for Protein Pattern Using Fuzzy Integral (퍼지적분을 이용한 단백질패턴에 관한 특징추출)

  • Song, Young-Jun;Kwon, Heak-Bong;Kim, Mi-Hye
    • The Journal of the Korea Contents Association
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    • v.7 no.1
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    • pp.40-47
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    • 2007
  • In the protein macro array image, it is important to find out the feature of the each protein chip. A decision error by the personal sense of sight occurred from long time observation while making an experiment in many protein chip image. So the feature extraction is needed by a simulator. In the case of feature analysis for macro array scan image the efficiency is maximized. In the fluorescence scan image, the response for each cell have been depend on R, G, B distribution of color image. But it is difficult to be classified as one color feature in the case of mixed color image. In this paper, the response color of a protein chip is classified according to the fuzzy integral value with respect to fuzzy measure as the user desired color. The result of the experiment for the macro array fluorescence image with the Scan Array 5000 shows that the proposed method using the fuzzy integral is important fact to be make decision for the ambiguous color.

A Simulation of Advanced Multi-dimensional Isotachophoretic Protein Separation for Optimal Lab-on-a-chip Design (최적화된 Lab-on-a-chip 설계를 위한 향상된 다차원 프로틴 등속영동 시뮬레이션)

  • Cho, Mi-Gyung
    • Journal of the Korea Institute of Information and Communication Engineering
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    • v.13 no.7
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    • pp.1475-1482
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    • 2009
  • In this paper, a computer simulation is developed for isotachophoretic protein separation in a serpentine micro channel for optimal lab on a chip design using 2D Finite Element Method. This 2D ITP model is composed of 5 components such as hydrochloric acid as Leader, caproic acid as terminator, acetic acid and benzoic acid as two proteins, and histindine as background electrolyte. The computer model is based on mass conservation equation for 5 components, charge conservation equation for electric potential, and electro neutrality condition for pH calculation. For the validation of the 2D spatial ITP model, the results are compared with the Simul5 developed by Bohuslav Gas Group. The simulation results are in a good agreement in a ID planar channel. This proves the precision of our model. The 2Dproteinseparation is conducted in a 2D curved channel for Lab on a chip design and dispersions of proteins are revealed during the electrophoretic process in a curved shape.

Inference of Genetic Regulatory Modules Using ChIP-on-chip and mRNA Expression Data

  • Cho, Hye-Young;Lee, Do-Heon
    • Bioinformatics and Biosystems
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    • v.2 no.2
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    • pp.62-65
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    • 2007
  • We present here the strategy of data integration for inference of genetic regulatory modules. First, we construct all possible combinations of regulators of genes using chromatin-immunoprecipitation(ChIP)-chip data. Second, hierarchical clustering method is employed to analyze mRNA expression profiles. Third, integration method is applied to both of the data. Finally, we construct a genetic regulatory module which is involved in the function of ribosomal protein synthesis.

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