• Title/Summary/Keyword: Microarray chip

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Manufacturing Protein-DNA Chip for Depigmenting Agent Screening (전사인자 저해제 통한 미백제 탐색용 단백질 칩 제작)

  • Han Jung-Sun;Kwak Eun-Young;Lee Hyang-Bok;Shin Jlung-Hyun;Baek Seung-Hak;Chung Bong-Hyun;Kim Eun-Ki
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.30 no.4 s.48
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    • pp.479-483
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    • 2004
  • An attempt was made to develop a proteinchip for screening of MITF (microphthalmia transcription factor) inhibitor. Binding of MITF to E-box causes transcription of several pigmenting genes including tyrosinase gene. We investigated binding of MITF and its DNA binding site (E-box) using a protein-DNA chip with various detection methods including flurorescence (Cyt3). SPR (surface plasmon resonance) and SPRi (surface plasmon resonance imaging). A fusion protein (MITF-Maltose Binding Protein) was attached on the glass plate by chemical modification. An inhibitory synthetic DNA oligomer, artificially designed based on the E-box sequence, inhibited the binding of MITF and E-box. These results showed the potentials of flurorescence-based MITF protein chip as a microarray for high throughput screening (HTS) system of depigmenting agents.

Nonstandard Machine Learning Algorithms for Microarray Data Mining

  • Zhang, Byoung-Tak
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2001.10a
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    • pp.165-196
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    • 2001
  • DNA chip 또는 microarray는 다수의 유전자 또는 유전자 조각을 (보통 수천내지 수만 개)칩상에 고정시켜 놓고 DNA hybridization 반응을 이용하여 유전자들의 발현 양상을 분석할 수 있는 기술이다. 이러한 high-throughput기술은 예전에는 생각하지 못했던 여러가지 분자생물학의 문제에 대한 해답을 제시해 줄 수 있을 뿐 만 아니라, 분자수준에서의 질병 진단, 신약 개발, 환경 오염 문제의 해결 등 그 응용 가능성이 무한하다. 이 기술의 실용적인 적용을 위해서는 DNA chip을 제작하기 위한 하드웨어/웻웨어 기술 외에도 이러한 데이터로부터 최대한 유용하고 새로운 지식을 창출하기 위한 bioinformatics 기술이 핵심이라고 할 수 있다. 유전자 발현 패턴을 데이터마이닝하는 문제는 크게 clustering, classification, dependency analysis로 구분할 수 있으며 이러한 기술은 통계학과인공지능 기계학습에 기반을 두고 있다. 주로 사용된 기법으로는 principal component analysis, hierarchical clustering, k-means, self-organizing maps, decision trees, multilayer perceptron neural networks, association rules 등이다. 본 세미나에서는 이러한 기본적인 기계학습 기술 외에 최근에 연구되고 있는 새로운 학습 기술로서 probabilistic graphical model (PGM)을 소개하고 이를 DNA chip 데이터 분석에 응용하는 연구를 살펴본다. PGM은 인공신경망, 그래프 이론, 확률 이론이 결합되어 형성된 기계학습 모델로서 인간 두뇌의 기억과 학습 기작에 기반을 두고 있으며 다른 기계학습 모델과의 큰 차이점 중의 하나는 generative model이라는 것이다. 즉 일단 모델이 만들어지면 이것으로부터 새로운 데이터를 생성할 수 있는 능력이 있어서, 만들어진 모델을 검증하고 이로부터 새로운 사실을 추론해 낼 수 있어 biological data mining 문제에서와 같이 새로운 지식을 발견하는 exploratory analysis에 적합하다. 또한probabilistic graphical model은 기존의 신경망 모델과는 달리 deterministic한의사결정이 아니라 확률에 기반한 soft inference를 하고 학습된 모델로부터 관련된 요인들간의 인과관계(causal relationship) 또는 상호의존관계(dependency)를 분석하기에 적합한 장점이 있다. 군체적인 PGM 모델의 예로서, Bayesian network, nonnegative matrix factorization (NMF), generative topographic mapping (GTM)의 구조와 학습 및 추론알고리즘을소개하고 이를 DNA칩 데이터 분석 평가 대회인 CAMDA-2000과 CAMDA-2001에서 사용된cancer diagnosis 문제와 gene-drug dependency analysis 문제에 적용한 결과를 살펴본다.

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Gene expression profiling of SH -SY5Y cells in neuroprotective effect of total ginsenosides on H202 induced neurotoxicity (인간 신경모세포종 SH-SY5Y에서 인삼(人蔘) total ginsenosides의 신경보호 기능에 관련된 유전자 발현 양상에 대한 연구)

  • Lee, Seung-Gi;Chai, Young-Gyu;Jung, Kyoung-Hwa;Kim, Ji-Hyouck;Hu, Yong-Suk
    • Journal of Oriental Neuropsychiatry
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    • v.18 no.1
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    • pp.95-110
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    • 2007
  • Objective : The purpose of this study was to investigate molecular basis of neuroprotective effect in total ginsenosides. After H202 induced neurotoxicity, gene expression profiling of SH-SY5Y neuroblastoma cells treated by total ginsenosides is analyzed. Method : After SH-SY5Y cells were cultured, they were damaged by H202 induced oxidative stress. After twenty four hours, experimental group is treated by total ginsenosides and control group is treated by 0.9% saline. A high density cDNA microarray chip is used to analyze the gene expression profiling of SH-SY5Y cells. The Significance Analysis of Microarray method is used for identifying genes on a microarray. Results : 1. According to the results of microarray experiment, 17 genes were up-regulated, 38 genes were down-regulated. 2. Expression of OPHNl, KTANl, ATM, PRKCE, MAPKs genes associated with cell proliferation, neural growth, and the prevention of apoptosis were increased. 3. Change of EPX gene was the greatest among all genes. EPX gene associated with oxidative stress, and tumor suppressor gene ADAM11 were decreased. Conclusion : According to this study, molecular basis of neuroprotective effect of total ginsenosides is as followings: the increase of gene expression associated with cell proliferation, neuron growth, the prevention of apoptotsis and decrease of gene expression associated with oxidative stress and tumor suppressor.

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Microarray Analysis of Differentially Expressed Genes between Cysts and Trophozoites of Acanthamoeba castellanii

  • Moon, Eun-Kyung;Xuan, Ying-Hua;Chung, Dong-Il;Hong, Yeon-Chul;Kong, Hyun-Hee
    • Parasites, Hosts and Diseases
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    • v.49 no.4
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    • pp.341-347
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    • 2011
  • Acanthamoeba infection is difficult to treat because of the resistance property of Acanthamoeba cyst against the host immune system, diverse antibiotics, and therapeutic agents. To identify encystation mediating factors of Acanthamoeba, we compared the transcription profile between cysts and trophozoites using microarray analysis. The DNA chip was composed of 12,544 genes based on expressed sequence tag (EST) from an Acanthamoeba ESTs database (DB) constructed in our laboratory, genetic information of Acanthamoeba from TBest DB, and all of Acanthamoeba related genes registered in the NCBI. Microarray analysis indicated that 701 genes showed higher expression than 2 folds in cysts than in trophozoites, and 859 genes were less expressed in cysts than in trophozoites. The results of real-time PCR analysis of randomly selected 9 genes of which expression was increased during cyst formation were coincided well with the microarray results. Eukaryotic orthologous groups (KOG) analysis showed an increment in T article (signal transduction mechanisms) and O article (posttranslational modification, protein turnover, and chaperones) whereas significant decrement of C article (energy production and conversion) during cyst formation. Especially, cystein proteinases showed high expression changes (282 folds) with significant increases in real-time PCR, suggesting a pivotal role of this proteinase in the cyst formation of Acanthamoeba. The present study provides important clues for the identification and characterization of encystation mediating factors of Acanthamoeba.

DNA microarray analysis of gene expression of MC3T3-E1 osteoblast cell cultured on anodized- or machined titanium surface

  • Park, Ju-Mi;Jeon, Hye-Ran;Pang, Eun-Kyoung;Kim, Myung-Rae;Kang, Na-Ra
    • Journal of Periodontal and Implant Science
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    • v.38 no.sup2
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    • pp.299-308
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    • 2008
  • Purpose: The aim of this study was to evaluate adhesion and gene expression of the MC3T3-E1 cells cultured on machined titanium surface (MS) and anodized titanium surface (AS) using MTT test, Scanning electron micrograph and cDNA microarray. Materials and Methods: The MTT test assay was used for examining the proliferation of MC3T3-E1 cells, osteoblast like cells from Rat calvaria, on MS and AS for 24 hours and 48 hours. Cell cultures were incubated for 24 hours to evaluate the influence of the substrate geometry on both surfaces using a Scanning Electron Micrograph (SEM). The cDNA microarray Agilent Rat 22K chip was used to monitor expressions of genes. Results: After 24 hours of adhesion, the cell density on AS was higher than MS (p < 0.05). After 48 hours the cell density on both titanium surfaces were similar (p > 0.05). AS had the irregular, rough and porous surface texture. After 48 hours incubation of the MC3T3-E1 cells, connective tissue growth factor (CTGF) was up-regulated on AS than MS (more than 2 fold) and the insulin-like growth factor 1 receptor was down-regulated (more than 2 fold) on AS than MS. Conclusion: Microarray assay at 48 hours after culturing the cells on both surfaces revealed that osteoinductive molecules appeared more prominent on AS, whereas the adhesion molecules on the biomaterial were higher on MS than AS, which will affect the phenotype of the plated cells depending on the surface morphology.

Web-based microarray analysis using the virtual chip viewer and bioconductor. (MicroArray의 직관적 시각적 분석을 위한 웹 기반 분석 도구)

  • Lee, Seung-Won;Park, Jun-Hyung;Kim, Hyun-Jin;Kang, Byeong-Chul;Park, Hee-Kyung;Kim, In-Ju;Kim, Cheol-Min
    • Proceedings of the Korea Inteligent Information System Society Conference
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    • 2005.05a
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    • pp.198-201
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    • 2005
  • DNA microarray 칩은 신약 개발, 유전적 질환 진단, Bio-molecular 상호작용 연구, 유전자의 기능연구 등 폭넓게 사용되고 있다. 이 논문은 cDNA mimcroarray 데이터를 분석하기 위한 웹형태의 시스템 개발에 대한 내용을 다룬다. 하나의 cDNA microarray에는 수 백에서 수 만개의 유전자가 심어져 있으며, 데이터를 분석할 때 대량의 데이터와 다양한 형태의 오류로 인해서 데이터간의 차이를 보정하는 분석 도구와 통계적 기법들이 사용되어야 한다. 본 논문에서는 가상 칩 뷰어를 이용하여 실제 microarray 데이터의 foreground intensity에서 백그라운드의 intensity를 제거하여 일반화된 칩 이미지를 생성한다. 이 가상 칩 뷰어는 여러 가지 필터효과와 서로 다른 두 형광의 차이를 조정하는 global normalization 기법을 사용하여 발현 유전자 분석을 시각적으로 할 수 있고, 중복된 마이크로어레이 칩 데이터를 통하여 시간이 많이 걸리는 분석전 칩의 유효성을 검토할 수 있다. 칩 데이터의 normalization을 위한 통계 방법으로 R 통계 도구와 linear 모델을 사용하여 microarray 칩의 유전자 발현 양상을 분석한다. 통계적 방법을 사용하지 않은 데이터를 추출, 이 데이터의 패턴 그래프 그리고 발현 레벨을 분류하여 마이크로어레이의 각 스팟의 유효성 검토의 정확성을 높였다. 이 시스템은 칩의 유효성 검토, 스팟의 유효성 검토, 유전자 선정에 대해 분석의 용이성과 정확성을 높일 수 있었다.

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Nonlinear matching measure for the analysis of on-off type microarray image (온-오프 형태의 DNA 마이크로어레이 영상 분석을 위한 비선형 정합도)

  • Ryu Mun ho;Kim Jong dae
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.30 no.3C
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    • pp.112-118
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    • 2005
  • In this paper, we propose a new nonlinear matching measure for automatic analysis of the on-off type DNA microarray images in which the hybridized spots are detected by the template matching method. The proposed measure is obtained by binary-thresholding over the whole template region and taking the number of white pixels inside the spotted area. This measure is compared with the normalized covariance in terms of the classification ability of the successfulness of the locating markers. The proposed measure is evaluated for the scanned images of HPV DNA microarrays where the marker locating is a critical issue because of the small number of spots. The targeting spots of HPV DNA chips are designed for genotyping 22 types of the human papilloma virus(HPV). The proposed measure is proven to give more discriminative response reducing the miss cases of the successful marker locating.

Comparative Transcriptome Analysis for Avermectin Overproduction via Streptomyces avermitilis Microarray System

  • Im, Jong-Hyuk;Kim, Myung-Gun;Kim, Eung-Soo
    • Journal of Microbiology and Biotechnology
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    • v.17 no.3
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    • pp.534-538
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    • 2007
  • Avermectin and its analogs are major commercial antiparasitic agents in the fields of animal health, agriculture, and human infections. To increase our understanding about the genetic mechanism underlying avermectin overproduction, comparative transcriptomes were analyzed between the low producer S. avermitilis ATCC31267 and the high producer S. avermitilis ATCC31780 via a S. avermitilis whole genome chip. The comparative transcriptome analysis revealed that fifty S. avermitilis genes were expressed at least two-fold higher in S. avermitilis ATCC31780. In particular, all the avermectin biosynthetic genes, including polyketide synthase (PKS) genes and an avermectin pathway-specific regulatory gene, were less expressed in the low producer S. avermitilis ATCC31267. The present results imply that avermectin overproduction in S. avermitilis ATCC31780 could be attributed to the previously unidentified fifty genes reported here and increased transcription levels of avermectin PKS genes.

Effects of an Extract from the Roots of Platycodon grandiflorum on the Growth and Gene Expression of Human Lung Carcinoma NCI-H460 Cells (DNA chip에 의한 연구에 따른 길경 수용액 추출물이 NCI-H460 인체 폐암세포의 성장 및 유전자 발현에 미치는 영향)

  • Kim, Hoon;Park, Dong-Il
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.19 no.2
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    • pp.77-87
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    • 2006
  • Objectives : We studied Effect of Platycodon grandiflorum on Lung carcinoma Methods : By using cDNA microarray technique, we analyzed the effects of AEPG(aqueous extract of Platycodon grandiflorum) on the gene expression profile. Results : Out of 384 genes screened associated with growth inhibition of carcinoma cell, 9 genes were founded to be affected in their expression levels by more than 1.2-fold after treatment with AEPG. And 67 genes were changed the expression level 0.5 times more than that of reference RNA after treatment of AEPG. Conclusions: These findings suggest that P. grandiflorum has strong potential for development as an agent for prevention and treatment against human lung cancer.

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Regulation of Pipernonaline on Biological Functions of Human Prostate Cancer Cells Based on Microarray Analysis (Microarray를 이용한 pipernonaline의 인간 전립선 암세포에 대한 기능 조절 분석)

  • Kim, Sang-Hun;Kim, Kwang-Youn;Yu, Sun-Nyoung;Park, Seul-Ki;Kwak, In-Seok;Rhee, Moon-Soo;Bang, Byung-Ho;Chun, Sung-Sik;Ahn, Soon-Cheol
    • Journal of Life Science
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    • v.22 no.11
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    • pp.1552-1557
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    • 2012
  • It has been reported that pipernonaline isolated from Piper longum Linn. has a wide biochemical and pharmacological effect, including antitumor activity in prostate cancer PC-3 cells. However, its mechanism and expression pattern of many genes involved in biological functions are not clearly understood. To perform the gene expression study in PC-3 cells treated with pipernonaline, a cDNA microarray chip composed of 44,000 human cDNA probes was used. As a result, cell cycle-related genes, apoptosis-related genes, and cell proliferation/growth-related genes have been identified in gene ontology of the DAVID database. These results suggest that pipernonaline has antitumor activity by regulating the expression pattern of genes involved in biological signaling pathway in prostate cancer PC-3 cells. Further, additional analysis of these microarray data can be a useful tool to identify the mechanism and discovery of novel genes in cancer therapy.