• Title/Summary/Keyword: cDNA microarray

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Identification of Biomarkers for Radiation Response Using cDNA Microarray

  • Park, Woong-Yang
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2001.10a
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    • pp.29-44
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    • 2001
  • DNA damage by physical insult including UV and g-radiation might provoke genetic alterations in cells, which is followed by either acute cell death or tumorigenesis. The responsiveness to g-radiation depends on cellular context of target cells. To understand the mechanisms of checkpoint control, repair and cell death following genotoxic stimu]i, cDNA microarray can provide the gene expression profile. To make a profile of gene expression in irradiated Jurkat T cells, we hybridized the cDNA microarray using cDNA from g-irradiated Jurkat T cells. Jurkat T cells were exposed to 4Gy to 16Gy, and total RNA were extracted at 4 to 24 hrs after irradiation. The hybridization of the microarray to fluorescence-labeled cDNA from treated and untreated cells was analyzed by bioinformatic analysis to address relative changes in expression levels of the genes present in the array. Responses varied widely in different time points, suggesting acute stress response and chronic restoration or cell death. From these results we could select 384 genes related to radiation response in Tcells, and radiation response might be different in various types of cells. Using Radchip, we could separate "the exposed" from control PBMCs. We propose that Radchip might be useful to check the radiation research as well as radiation carcinogenesis.

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Effects of Dioxin Exposed in Human by Using Radioactive cDNA Microarray

  • Ryu, Yeon-Mi;Kim, Ki-Nam;Kim, Hye-Won;Sohn, Sung-Hwa;Lee, Seung-Ho;Kim, Yu-Ri;Seo, Sang-Hui;Lee, Seung-Min;Lee, Eun-Il;Kim, Meyoung-Kon
    • Molecular & Cellular Toxicology
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    • v.2 no.1
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    • pp.35-47
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    • 2006
  • 2, 3, 7, 8-Tetrachlorodibenzo-p-dioxin (TCDD) are well known as the most toxic environmental compound in these days. Many researches are reported that dioxin produces multiple toxic effects, such as endocrine toxicity, reproductive toxicity, immunotoxicity and cancer. In this study, we carried to discover novel evidence for previously unknown gene expression patterns in human exposed to dioxin by using radioactive cDNA microarray. 548 workers who were divided into experimental and control groups according to their urinary Naphthol levels were enrolled in our study. Blood mRNA in human was isolated, and the gene expression profiles were analyzed by cDNA microarray. Gene expression analysis identified 52 genes which exhibited a significant change. In our study, most notably, genes involved in cell cycle, cell proliferation, signal transduction and apoptosis in human exposed to dioxin, such as CCND3, TSHR, and EFRN5, were up-regulated. In the current study, we observed gene expression of people that are exposed to dioxin using radioactive cDNA microarray. Through these results, we suggest when objects are exposed to toxic compounds, such as dioxin, the radioactive cDNA microarray may be using in sensitively detecting of cancerous change.

Comparison of Expression Profiling of Gastric Cancer by O1igonucleotide and cDNA Microarrays (O1igonucleotide Microarray와 cDNA Microarray를 이용한 위암조직의 대단위 유전자 발현 비교)

  • Jung, Kwang-Hwa;Kim, Jung-Kyu;Noh, Ji-Heon;Eun, Jung-Woo;Bae, Hyun-Jin;Lee, Sug-Hyung;Park, Won-Sang;Yoo, Nam-Jin;Lee, Jung-Young;Nam, Suk-Woo
    • YAKHAK HOEJI
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    • v.51 no.3
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    • pp.179-185
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    • 2007
  • Gastric cancer is one of the most common malignancies in Korea, but the predominant molecular event underlying gastric carcinogenesis remain unknown. Recently, DNA microarray technology has enabled the comprehensive analysis of gene expression level, and as such has yielded great insight into the molecular nature of cancer, However, despite the powerful approach of this techniques, the technical artifacts and/or bias in applied array platform limited the liability of resultant tens of thousand data points from microarray experiments. Therefore, we applied two different any platforms, such as olignucleotide microarray and cDNA microarray, to identify gastric cancer related large-scale molecular signature of the same human specimens. When thirty sets of matched human gastric cancer and normal tissues subjected to oligonucleotide microarray, total 623 genes were resulted as differently expressed genes in gastric cancer compared to normal tissues, and 252 genes for cDNA microarray analysis. In addition, forty three outlier genes which reflect the characteristic expression signature of gastric cancer beyond array platform and analytical protocol was recapitulated from two different expression profile. In conclusion, we were able to identify robust large-scale molecular changes in gastric cancer by applying two different platform of DNA microarray, this may facilitate to understand molecular carcinogenesis of gastric cancer.

Gene Expression study of human chromosomal aneuploid

  • Lee Su-Man
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2006.02a
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    • pp.98-107
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    • 2006
  • Chromosomal copy number changes (aneuploidies) are common in human populations. The extra chromosome can affect gene expression by whole-genome level. By gene expression microarray analysis, we want to find aberrant gene expression due to aneuploidies in Klinefelter (+X) and Down syndrome (+21). We have analyzed the inactivation status of X-linked genes in Klinefelter Syndrome (KS) by using X-linked cDNA microarray and cSNP analysis. We analyzed the expression of 190 X-linked genes by cDNA microarray from the lymphocytes of five KS patients and five females (XX) with normal males (XY) controls. cDNA microarray experiments and cSNP analysis showed the differentially expressed genes were similar between KS and XX cases. To analyze the differential gene expressions in Down Syndrome (DS), Amniotic Fluid (AF)cells were collected from 12 pregnancies at $16{\sim}18$ weeks of gestation in DS (n=6) and normal (n=6) subjects. We also analysis AF cells for a DNA microarray system and compared the chip data with two dimensional protein gel analysis of amniotic fluid. Our data may provide the basis for a more systematic identification of biological markers of fetal DS, thus leading to an improved understanding of pathogenesis for fetal DS.

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Gene Expression in Zn-deficient U937 Cell Line : Using cDNA Microarray (아연결핍된 단핵구 U937 Cell Line에 있어서의 유전자 발현 탐색 : cDNA Microarray 기법 이용)

  • Beattie, John H.;Trayhurn, Paul
    • Journal of Nutrition and Health
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    • v.35 no.10
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    • pp.1053-1059
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    • 2002
  • In post-genome period, the technique for identifying gene expression has been changed to high throughput screening. In the field of molecular nutrition, the need for this technique to clarify molecular function of the specific nutrient is essential. In this study, we have tested the zinc-regulated gene expression in zinc-deficient U937 cells, using cDNA microarray which is the cutting-edge technique to screen large numbers of gene expression simultaneously. The study result can be used for the preliminary gene screening data for clarifying, using monocyte U937 cell line, molecular Zn aspect in atherosclerosis. U937 cells were cultured in Zn-adequate (control, 12 $\mu$M Zn) or Zn-deficient (experimental, 0 $\mu$M Zn) ESMI media during 2 days, respectively. Cells were harvested and RNA was extracted. Total RNA was reverse-transcriptinized and synthesized cDNA probe labeled with Cy-3. fluorescent labeled cDNA probe was applied to microarray slide for hybridization slide, and after then, the slide was scanned using fluorescence scanner. ‘Highly expressed genes’ in Zn-deficient U937 cells, comparing to Zn-adequate group, are mainly about the genes for motility protein, immune system protein, oncogene and tumor suppressor and ‘Less highly expressed genes’ are about the genes for transcription, apoptosis associated protein, cell cycle, and several basic transcription factors. The results of this preliminary study imply the effectiveness of cDNA microarray for expression profiling of a singly nutrient deficiency, specially Zn. Furthur study, using tailored-cDNA array and capillary endothelial cell lines, would be beneficial to clarify molecular Zn function, more in detail.

Identification of Virulence Factors in Vibrio vulnificus by Comparative Transcriptomic Analyses between Clinical and Environmental Isolates Using cDNA Microarray

  • Kim, In-Hwang;Kim, Byung-Soo;Lee, Kyung-Shin;Kim, Ik-Joong;Son, Jee-Soo;Kim, Kun-Soo
    • Journal of Microbiology and Biotechnology
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    • v.21 no.12
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    • pp.1228-1235
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    • 2011
  • We compared the gene expression among four clinical and five environmental V. vulnificus isolates, using a cDNA microarray containing 131 genes possibly associated with pathogenicity, transport, signal transduction, and gene regulations in the pathogen. cDNAs from total RNAs of these isolates were hybridized into the cDNA microarray using the cDNA of the wild-type strain MO6-24/O as a reference. We focused on selecting differentially expressed (DE) genes between clinical and environmental isolates using a modified t-statistic. We could detect two statistically significant DE genes between virulent isolates and less-virulent isolates with a marginal statistical significance (p-value of 0.008). These were genes putatively encoding pilin and adenlyate cylase. Real time-PCR confirmed that these two selected genes transcribed in significantly higher levels in virulent isolates than in less-virulent isolates. Mutants with lesions in the gene encoding pilin showed significantly higher $LD_{50}$ values than that of wild type.

cDNA Microarray Normalization에 대한 연구

  • Kim, Jong-Yeong;Lee, Jae-Won
    • Proceedings of the Korean Statistical Society Conference
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    • 2003.10a
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    • pp.331-334
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    • 2003
  • 마이크로 어레이(microarray)실험에서 표준화(normalization)는 유전자의 발현수준에 영향을 미치는 여러 기술적인 변인을 제거하는 과정이다. cDNA microarray normalization에 있어 여러 방법이 제안되었지만, 이중 print-tip 효과가 존재할 때 사용되는 방법으로 print-tip lowess normalization이 대표적으로 사용된다. normalization에 사용되는 lowess 함수는 데이터의 특성에 따라 window width를 정해야만 연구의 목적에 맞는 결과를 도출할 수 있다. 본 논문에서는 각각의 tip에서 최적의 window width를 계산하는 절차를 논의하였다. 또한 이의 결과와 기존의 같은 window width를 사용하는 print-tip lowess normalization 결과와 비교 평가하여 normalization의 기본 원칙에 대한 타당성을 확인하였다.

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Comparison of Hybridization Behavior between Double and Single Strand of Targets and the Application of Asymmetric PCR Targets in cDNA Microarray

  • Wei, Qing;Liu, Sanzhen;Huang, Jianfeng;Mao, Xueying;Chu, Xiaohui;Wang, Yu;Qiu, Minyan;Mao, Yumin;Xie, Yi;Li, Yao
    • BMB Reports
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    • v.37 no.4
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    • pp.439-444
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    • 2004
  • Double stranded targets on the cDNA microarray contain representatives of both the coding and noncoding strands, which will introduce hybridization competition with probes. Here, the effect of double and single strands of targets on the signal intensity and the ratios of Cy5/Cy3 within the same slide were compared. The results show that single stranded targets can increase the hybridization efficiency without changing the Cy5/Cy3 ratio. Based on these results, a new strategy was established by generating cDNA targets with asymmetric PCR, instead of conventional PCR, to increase the sensitivity of the cDNA microarray. Furthermore, the feasibility of this approach was validated. The results indicate that the cDNA microarray system based on asymmetric PCR is more sensitive, with no decrease in the reliability and reproducibility as compared with that based on conventional symmetric PCR.

cDNA microarray profiling of Bombyx mori(kl20) during early embryogenesis

  • Hong, Sun-Mee;Kang, Seok-Woo;O, Tae-Jaeng;Kim, Nam-Soon;Lee, Jin-Sung;Goo, Tae-Won;Yun, Eun-Young;Choi, Ho;Hwang, Jae-Sam;Nho, Si-Kab
    • Proceedings of the Korean Society of Sericultural Science Conference
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    • 2003.04a
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    • pp.47-48
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    • 2003
  • The development of cDNA microarray has permitted the analysis of thousands of genes simultaneously. cDNA microarray has been used to analyze gene expression profiles during developmental stage in both single and multicellular organisms. Two significant factors contributing to the limitation of the development of cDNA microarray in the Bombyx mori are the shortage of accessible repositories of cDNA clones and ESTs and the relative scarcity of facilities to produce microarrays and analyze the data generated. (omitted)

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Toxicogenomic analysis of Effects of Bisphenol A on Japanese Medaka fish using high density-functional cDNA microarray

  • Jiho Min;Park, Kyeong-Seo;Hong, Han-Na;Gu, Man-Bock
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 2003.10a
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    • pp.173-173
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    • 2003
  • With the introduction of DNA microarrays, a high throughput analysis of gene expression is now possible as a replacement to the traditional time-consuming Southern-blot analysis. This cDNA microarray should be ahighly favored technology in the area of molecular toxicology or analysis of environmental stresses.In this study, therefore, we developed a novel cDNA microarray for analyzing stress-specific responses in japanese Medaka fish. In the design and fabrication of this stress specific functional cDNA microarray, 123 different genes in Medaka fish were selected from eighteen different stress responsive groups and spotted on a 25${\times}$75 mm glass surface. After exposure of the fish to bisphenol A which is the one of the well-known endocrine disrupting chemicals (EDCs), over 1 or 10 days, the responses of the DNA chip were found to show distinct expression patterns according to the mode of toxic actions from environmental toxicants. As a results, they showed specific gene expression pattern to bisphenol A, additionally, the chemical spesific biomarkers could be suggested based on the chip analysis data. Therefore, this chip can be used to monitor stress responses of unknown and/or known toxic chemicals using Medaka fish and may be used for the further development of biomarkers by utilizing the gene expression patterns for known contaminants.

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