• Title/Summary/Keyword: cDNA microarray chip

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The Antioxidant Effect, Inhibition of Interleukin-4 and the Effect on the Gene Expression by Using cDNA Chip of Chungsangboha-tang(Qingshangbuxia-tang) (청상보하탕의 항산화 효과, Interleukin-4 억제 및 cDNA chip을 이용한 유전자발현에 미치는 영향)

  • 이동생;정희재;정승기;이형구
    • The Journal of Korean Medicine
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    • v.24 no.2
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    • pp.148-158
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    • 2003
  • Backgrounds & Objectives: In many recent studies, molecular biological methods have been used to investigate the role of cytokines in pathogenesis and new therapeutic targets of asthma. Recently, as a method of research on the gene expression, they are applying another method which assays multiple gene expressions at the same time by the microarray. In this study, the antioxidant effect, the inhibitory effect against interleukin-4 and the effect on the CD/cytokine gene expression in PBMC (peripheral blood mononuclear cells) was evaluated by using cDNA microarray chip of Chungsangboha-tang. Methods: Experimental studies were performed for the antioxidant effect of Chungsangboha-tang on DPPH (1, 1-diphenyl-2-picrylhydrazyl) solution, for the IL-4-inhibiting effect on BALB/c mouse spleen, and for the gene expression effect on PBMC (peripheral blood mononuclear cells) with microarray. Results: Chungsangboha-tang showed antioxidant effect dose-dependently. Chungsangboha-tang inhibited interleukin-4 dose-dependently and showed significant difference in 10ug/ml and 100ug/ml of test groups. There was no 2 more times upregulated genes than in the control group by using cDNA microarray chip of Chungsangbohn-tang, but there were 140%-200% upregulated genes. There was no 2 more times downregulated genes than in the control group by using cDNA microarray chip of Chungsangboha-Tang, but there was 50%-75% downregulated genes. Conclusions: This study showed that Chungsangboha-tang has an antioxidant effect and inhibition of Interleukin-4, but further studies are necessary with microarray.

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Highly Integrated DNA Chip Microarrays by Hydrophobic Interaction

  • Park, Yong-Sung;Kim, Do-Kyin;Kwon, Young-Soo
    • KIEE International Transactions on Electrophysics and Applications
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    • v.11C no.2
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    • pp.23-27
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    • 2001
  • Microarray-based DNA chips provide an architecture for multi-analyte sensing. In this paper, we report a new approach for DNA chip microarray fabrication. Multifunctional DNA chip microarrays were made by immobilizing many kinds if DNAs on transducers (particles). DNA chip microarrays were prepared by randomly distributing a mixture of the particles on a chip pattern containing thousands of micro meter-scale sites. The particles occupied different sites from array to array. Each particle cam be distinguished by a tag that is established on the particle. The particles were arranged on the chip pattern by the random fluidic self-assembly (RFSA) method, using hydrophobic interaction.

Development of High-Intergrated DNA Array on a Microchip by Fluidic Self-assembly of Particles (담체자기조직화법에 의한 고집적 DNA 어레이형 마이크로칩의 개발)

  • Kim, Do-Gyun;Choe, Yong-Seong;Gwon, Yeong-Su
    • The Transactions of the Korean Institute of Electrical Engineers C
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    • v.51 no.7
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    • pp.328-334
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    • 2002
  • The DNA chips are devices associating the specific recognition properties of two DNA single strands through hybridization process with the performances of the microtechnology. In the literature, the "Gene chip" or "DNA chip" terminology is employed in a wide way and includes macroarrays and microarrays. Standard definitions are not yet clearly exposed. Generally, the difference between macro and microarray concerns the number of active areas and their size, Macroarrays correspond to devices containing some tens spots of 500$\mu$m or larger in diameter. microarrays concern devices containing thousnads spots of size less than 500$\mu$m. The key technical parameters for evaluating microarray-manufacturing technologies include microarray density and design, biochemical composition and versatility, repreducibility, throughput, quality, cost and ease of prototyping. Here we report, a new method in which minute particles are arranged in a random fashion on a chip pattern using random fluidic self-assembly (RFSA) method by hydrophobic interaction. We intend to improve the stability of the particles at the time of arrangement by establishing a wall on the chip pattern, besides distinction of an individual particle is enabled by giving a tag structure. This study demonstrates the fabrication of a chip pattern, immobilization of DNA to the particles and arrangement of the minute particle groups on the chip pattern by hydrophobic interaction.ophobic interaction.

Construction of Ovine Customer cDNA Chip and Analysis of Gene Expression Patterns in the Muscle and Fat Tissues of Native Korean Cattle (cDNA microarray를 이용하여 한우의 근육과 지방조직의 유전자 발현 패턴 분석 및 bovine customer cDNA chip 구성 연구)

  • Han, Kyung Ho;Choi, Eun Young;Hong, Yeon-Hee;Kim, Jae Yeong;Choi, In Soon;Lee, Sang-Suk;Choi, Yun Jaie;Cho, Kwang Keun
    • Journal of Life Science
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    • v.25 no.4
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    • pp.376-384
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    • 2015
  • To investigate the molecular events of controlling intramuscular fat (or marbling), which is an important factor in the evaluation of beef quality, we performed cDNA microarray analyses using the longissimus dorsi muscle and back fat tissues. For this study, we constructed normalized cDNA libraries: fat tissues in native Korean cattle (displaying 1,211 specific genes), and muscle tissues in native Korean cattle (displaying 1,346 specific genes). A bovine cDNA chip was constructed with 1,680 specific genes, consisting of 760 genes from muscle tissues and 920 genes from fat tissues. The microarray analysis in this experiment showed a number of differentially expressed genes, which compared the longissimus dorsi muscle (Cy5) with back fat tissue (Cy3). Among many specific differentially expressed genes, 12-lipoxygenase (oxidizing esterified fatty acids) and prostaglandin D synthase (differentiation of fibroblasts to adipocytes) are the key candidate enzymes that should be involved in controlling the accumulation of intramuscular fat. In this study, differentially and commonly expressed genes in the muscle and fat tissues of native Korean cattle were found in large numbers, using the hybridization assay. The expression levels of the selected genes were confirmed by semi-quantitative RT-PCR, and the results were similar to those of the cDNA microarray.

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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Electrochemical Detection of Single Nucleotide Polymorphism (SNP) Using Microelectrode Array on a DNA Chip (미소전극어레이형 DNA칩을 이용한 유전자다형의 전기화학적 검출)

  • 최용성;권영수;박대희
    • The Transactions of the Korean Institute of Electrical Engineers C
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    • v.53 no.5
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    • pp.286-292
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    • 2004
  • In this study, an integrated microelectrode array was fabricated on glass slide using microfabrication technology. Probe DNAs consisting of mercaptohexyl moiety at their 5-end were spotted on the gold electrode using micropipette or DNA arrayer utilizing the affinity between gold and sulfur. Cyclic voltammetry in 5mM ferricyanide/ferrocyanide solution at 100 ㎷/s confirmed the immobilization of probe DNA on the gold electrodes. When several DNAs were detected electrochemically, there was a difference between target DNA and control DNA in the anodic peak current values. It was derived from specific binding of Hoechst 33258 to the double stranded DNA due to hybridization of target DNA. It suggested that this DNA chip could recognize the sequence specific genes. It suggested that multichannel electrochemical DNA microarray is useful to develop a portable device for clinical gene diagnostic System.

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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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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Microarray 자료분석에서 표준화

  • 이성곤;박태성;최호식
    • Proceedings of the Korean Statistical Society Conference
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    • 2001.11a
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    • pp.149-153
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    • 2001
  • 본 논문은 microarray를 분석하기위한 표준화에 대한 여러 방법들을 소개하고 비교해보았다. Microarray 연구는 Human Genome Project에서 파생된 여러 생명공학 기술 중 가장 널리 사용되는 기술로 기존에는 하지 못했던 총체적인 유전자의 발현상황을 탐색할 수 있다는 장점을 지니고 있으나, 자료들에 일정한 패턴이 나타나거나 잡음이 첨가되어 정보의 추출이 용의하지 않다는 단점을 지니고 있다. 특히 자료에 일정한 패턴이 있는 경우에 올바르지 못한 결론을 이끌어낼 수도 있기에 이 패턴을 제거하는 표준화작업은 microarray 분석에 있어서 매우 중요한 처리과정이다. 본 논문에서는 표준화방법들을 소개하고 각각 가지고 있는 장단점을 실제 국내에서 얻어진 자료를 통해 비교하였고, 그 결과 LOWESS 적합을 통한 표준화방법이 타 방법에 비해 유용한 점이 많음을 확인할 수 있었다.

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A Method for Evaluation of the Quality of DNA Microarray Spots

  • Zhang, Bao;Ma, Wen-Li;Hu, Zi-You;Shi, Rong;Song, Yan-Bin;Zheng, Wen-Ling
    • BMB Reports
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    • v.35 no.5
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    • pp.532-535
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    • 2002
  • To establish a method to evaluate the quality of the printed microarray and DNA fragments' immobilization. The target gene fragments that were made with the restriction display PCR (RD-PCR) technique were printed on a superamine modified glass slide, then immobilized with UV cross-linking and heat. This chip was hybridized with universal primers that were labeled with cy3-dUTP, as well as cDNA that was labeled with cy3-dCTP, as the conventional protocol. Most of the target gene fragments on the chip showed positive signals, but the negative control showed no signal, and vice versa. We established a method that enables an effective evaluation of the quality of the microarrays.