• Title/Summary/Keyword: DNA microarray

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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.

A Probe Design Method for DNA Microarrays Using ${\epsilon}$-Multiobjetive Evolutionary Algorithms (${\epsilon}$-다중목적 진화연산을 이용한 DNA Microarray Probe 설계)

  • Cho Young-Min;Shin Soo-Yong;Lee In-Hee;Zhang Byoung-Tak
    • Proceedings of the Korean Information Science Society Conference
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    • 2006.06a
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    • pp.82-84
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    • 2006
  • 최근의 생물학적인 연구에 DNA microarray가 널리 쓰이고 있기 때문에, 이러한 DNA microarray를 구성하는데 필요한 probe design 작업의 중요성이 점차 커져가고 있다. 이 논문에서는 probe design 문제를 thermodynamic fitness function이 2개인 multi-objective optimization 작업으로 변환한 뒤, ${\epsilon}$-multiobjective evolutionary algorithm을 이용하여 probe set을 찾는다. 또한, probe 탐색공간의 크기를 줄이기 위하여 각 DNA sequence의 primer 영역을 찾는 작업을 진행하며, 사용자가 직접 프로그램을 테스트할 수 있는 웹사이트를 제공한다. 실험 대상으로는 mycoides를 선택하였으며, 이 논문에서 제안된 방법을 사용하여 성공적으로 probe set을 발견할 수 있었다.

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Oligonucleotide Probe Selection using Evolutionary Computation in Large Target Genes (다수의 목표 유전자에서 진화연산을 이용한 Oligonucleotide Probe 선택)

  • Shin, Ki-Roo;Kim, Sun;Zhang, Byung-Tak
    • Proceedings of the Korean Information Science Society Conference
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    • 2003.04c
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    • pp.455-457
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    • 2003
  • DNA microarray는 분자생물학에서 널리 사용되고 있는 실험 도구로써 크게 cDNA와 oligonucleotide microarray로 나뉘어진다. DNA microarray는 일련의 DNA 서열로 이루어진 probe들의 집합으로 구성되며 알려지지 않은 서열과의 hybridization 과정을 통해 특정 서열을 인식할 수 있게 된다. O1igonucieotide microarray는 cDNA 방법과는 다르게 probe를 구성하는 서열을 제작자가 임의로 구성할 수 있기 때문에 목표 서열이 가지는 고유한 부분만을 probe 서열로 사용함으로써 비용절감과 실험의 정확도를 높일 수 있다는 장점이 있다. 그러나 현재 목표 유전자 서열에 대해 probe 집합을 생성하는 결정적인 방법은 존재하지 않으며, 따라서 넓은 해 공간에서 효과적으로 최적 해를 찾아 주는 진화 연산이 probe 선택을 위한 좋은 대안으로 사용될 수 있다[1.2]. 그러나 진화연산을 이용한 probe 선택방법에 있어서 인식하고자 하는 목표 서열의 개수가 많아질 경우, 해 공간의 크기가 커짐으로 인해 문제점이 발생할 수 있다. 따라서 본 논문에서는 다수의 목표 유전자 서열을 대상으로 한 probe 선택 방법에 일어서 보다 효율적인 진화연산 접근 방법을 소개한다. 제시된 방법은 인식하고자 하는 목표 서얼의 일부를 선택해 이를 probe 집합의 후보로 사용하며. 유전 연산자를 이용한 진화과정을 통해 최적에 가까운 probe 집합을 찾는다. 본 논문은 GenBank로부터 유전자 서열을 대상으로 제안된 방법을 실험하였으며, 축소된 목표 서열만을 이용해 probe 집합을 선택하더라도 적합한 probe 집합을 찾을 수 있었다.

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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.

Gene Expression Analysis Using cDNA Microarray Assay by Cervi Pantotricuhum Cornu Herbal Acupuncture (녹용약침액(鹿茸藥鍼液)의 DNA chip을 이용(利用)한 유전자(遺傳子) 발현(發顯) 분석(分析))

  • Han, Sang-won;Seo, Jung-chul;Lee, Yun-ho;Choi, Je-yong
    • Journal of Acupuncture Research
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    • v.20 no.3
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    • pp.34-44
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    • 2003
  • Objective : Bone homeostasis is maintained by balance of bone formation and resorption. Therefore, bone related diseases arose by disturbance of this balance between osteoblast and osteoclast activities. To develop a successful screening system the therapeutic components based on oriental medicine is essential to set up systematic approach for that purpose. The purpose of this study is to the know the gene expression using cDNA microarray assay. Methods : Cervi Pantotricuhum Cornu Herbal-acupuncture extract was prepared by boiling. human osteosarcoma cells(HOS) were treated with Cervi Pantotricuhum Cornu Herbal-acupuncture solution. Then mRNA was extracted and cDNA microarray assay was performed. Results : Human osteosarcoma cells(HOS) treated with Cervi Pantotricuhum Cornu Herbal-acupuncture solution($500{\mu}g/m{\ell}$) showed that thioredoxin, TAFII31 and two novel genes were increased. However many genes decreased their expression by Cervi Pantotricuhum Cornu Herbal-acupuncture. Conclusions : This type of approach will give a good chance to explore the favorable effects of Cervi Pantotricuhum Cornu Herbal-acupuncture. Further study is needed for investigating the effect of Cervi Pantotricuhum Cornu Herbal-acupuncture.

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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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Genomic and Proteomic Profiling of the Cadmium Cytotoxic Response in Human Lung Epithelial Cells

  • Choi, Kwang-Man;Youn, Hyung-Sun;Lee, Mi-Young
    • Molecular & Cellular Toxicology
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    • v.5 no.3
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    • pp.198-206
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    • 2009
  • Microarray and proteomic expression patterns in response to cadmium exposure were analyzed in human lung epithelial cells. Among 35,000 genes analyzed by cDNA microarray, 228 genes were up-regulated and 99 genes were down-regulated, based on a fold change cut-off value of ${\geq}2$. Combining two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization-time-of-flight-mass spectrometry (MALDI-ToF-MS), 25 of 629 protein spots showed fold changes in expression ${\geq}2$ (17 up-regulated, 8 down-regulated). After comparing the cDNA microarray and proteomic analyses, only transglutaminase 2, translation elongation factor 1 alpha 1, and glyceraldehyde-3-phosphate dehydrogenase showed overlapping signals in the cDNA microarray and proteomic analyses, whereas the remaining differentially expressed proteins showed large discrepancies with respect to mRNA expression.

Basic Concept of Gene Microarray (Gene Microarray의 기본개념)

  • Hwang, Seung Yong
    • Korean Journal of Biological Psychiatry
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    • v.8 no.2
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    • pp.203-207
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    • 2001
  • The genome sequencing project has generated and will continue to generate enormous amounts of sequence data including 5 eukaryotic and about 60 prokaryotic genomes. Given this ever-increasing amounts of sequence information, new strategies are necessary to efficiently pursue the next phase of the genome project-the elucidation of gene expression patterns and gene product function on a whole genome scale. In order to assign functional information to the genome sequence, DNA chip(or gene microarray) technology was developed to efficiently identify the differential expression pattern of independent biological samples. DNA chip provides a new tool for genome expression analysis that may revolutionize many aspects of biotechnology including new drug discovery and disease diagnostics.

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Development of Microarrayer for DNA Chips (DNA Chip 제작을 위한 Microarrayer의 개발)

  • Kim, Suk-Yoel;Jung, Nam-Su;Im, Jae-Sung;Kim, Sang-Bong
    • Proceedings of the KSME Conference
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    • 2003.04a
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    • pp.899-904
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    • 2003
  • Microarrayer makes DNA chip and microarray that contain hundreds to thousands of immobilized DNA probes on surface of a microscope slide. This paper shows the development results for a printing type of microarrayer. It realizes a typical, low-cost and efficient microarrayer for generating low density microarray. The microarrayer is developed by using a robot of three-axes perpendicular type. It is composed of a computer-controlled three-axes robot and a pen tip assembly. The key component of the arrayer is the print-head containing the tips to immobilize cDNA, genomic DNA or similar biological material on glass surface. The robot is designed to automatically collect probes from two 96-well plates with up to 32 tips at the same time. To prove the performance of the developed microarrayer, the general water types of inks such as black, blue and red. The inks are distributed at proper positions of 96 well plates and the three color inks are immobilized on the slide glass under the operation procedure. As the result of the test, it can be shown that it has sufficient performance for the production of low integrated DNA chip consisted of 96 spots within 1 $cm^2$ area.

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Application of DNA Microarray Technology to Molecular Microbial Ecology

  • Cho Jae-Chang
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2002.10a
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    • pp.22-26
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    • 2002
  • There are a number of ways in which environmental microbiology and microbial ecology will benefit from DNA micro array technology. These include community genome arrays, SSU rDNA arrays, environmental functional gene arrays, population biology arrays, and there are clearly more different applications of microarray technology that can be applied to relevant problems in environmental microbiology. Two types of the applications, bacterial identification chip and functional gene detection chip, will be presented. For the bacterial identification chip, a new approach employing random genome fragments that eliminates the disadvantages of traditional DNA-DNA hybridization is proposed to identify and type bacteria based on genomic DNA-DNA similarity. Bacterial genomes are fragmented randomly, and representative fragments are spotted on a glass slide and then hybridized to test genomes. Resulting hybridization profiles are used in statistical procedures to identify test strains. Second, the direct binding version of microarray with a different array design and hybridization scheme is proposed to quantify target genes in environmental samples. Reference DNA was employed to normalize variations in spot size and hybridization. The approach for designing quantitative microarrays and the inferred equation from this study provide a simple and convenient way to estimate the target gene concentration from the hybridization signal ratio.

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