• Title/Summary/Keyword: Oligo DNA microarray

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Microarray Probe Design with Multiobjective Evolutionary Algorithm (다중목적함수 진화 알고리즘을 이용한 마이크로어레이 프로브 디자인)

  • Lee, In-Hee;Shin, Soo-Yong;Cho, Young-Min;Yang, Kyung-Ae;Zhang, Byoung-Tak
    • Journal of KIISE:Software and Applications
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    • v.35 no.8
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    • pp.501-511
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    • 2008
  • Probe design is one of the essential tasks in successful DNA microarray experiments. The requirements for probes vary as the purpose or type of microarray experiments. In general, most previous works use the simple filtering approach with the fixed threshold value for each requirement. Here, we formulate the probe design as a multiobjective optimization problem with the two objectives and solve it using ${\epsilon}$-multiobjective evolutionary algorithm. The suggested approach was applied in designing probes for 19 types of Human Papillomavirus and 52 genes in Arabidopsis Calmodulin multigene family and successfully produced more target specific probes compared to well known probe design tools such as OligoArray and OligoWiz.

Probe Selection of DNA Microarrays Using Genetic Algorithms (유전 알고리즘을 이용한 DNA Microarray의 Probe 선택)

  • Kim, Sun;Zhang, Byoung-Tak
    • Proceedings of the Korean Institute of Intelligent Systems Conference
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    • 2002.05a
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    • pp.183-187
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    • 2002
  • DNA microarray는 분자생물학 및 DNA 컴퓨팅 분야에 널리 사용되고 있는 실험 도구이다. DNA microarray를 이용하는 한 예는 알려진 유전자 집합을 바탕으로 하여 hybridization을 통해 새로운 DNA 서열을 분석하는 것이다. 이를 위한 가장 간단한 방법은 알려진 유전자의 모든 서열을 DNA microarray 상에 올려놓는 것이지만 이는 결과의 정확도 및 칩 제작비용 면에서 비효율적이다. 따라서 일반적으로는 유전자 서열 정보를 파악한 후 일련의 DNA 서열을 선택하는 probe 디자인 과정을 거친다. 그러나 현재 유전자 서열을 바탕으로 최적의 probe 집합을 찾는 결정적인 방법이 존재하고 있지 않다. 이에 본 논문은 oligo DNA microarray을 이용한 DNA 서열 분석 문제에 있어서 가능한 많은 유전자를 인식하면서 최소의 probe 개수를 갖는 집합을 찾는 방법을 제안한다. 제시된 방법은 가능한 probe 집합들로 해집합을 구성한 후, 유전알고리즘을 이용한 진화 과정을 통해 목적하는 probe 집합을 찾는다. 본 논문에서는 GenBank로부터 얻은 일련의 유전자 집합을 대상으로 실험하였으며 그 결과를 분석하였다.

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Genes expression monitoring using cDNA microarray: Protocol and Application

  • Muramatsu Masa-aki
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2000.11a
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    • pp.31-41
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    • 2000
  • The major issue in the post genome sequencing era is determination of gene expression patterns in variety of biological systems. A microarray system is a powerful technology for analyzing the expression profile of thousands of genes at one experiment. In this study, we constructed cDNA microarray which carries 2,304 cDNAS derived from oligo-capped mouse cDNA library. Using this hand-made microarray we determined gene expression in various biological systems. To determine tissue specific genes, we compared Nine genes were highly-expressed in adult mouse brain compared to kidney, liver, and skeletal muscle. Tissue distribution analysis using DNA microarray extracted 9 genes that were predominantly expressed in the brain. A database search showed that five of the 9 genes, MBP, SC1, HiAT3, S100 protein-beta, and SNAP25, were previously known to be expressed at high level in the brain and in the nervous system. One gene was highly sequence similar to rat S-Rex-s/human NSP-C, suggesting that the gene is a mouse homologue. The remaining three genes did not match to known genes in the GenBank/EMBL database, indicating that these are novel genes highly-expressed in the brain. Our DNA microarray was also used to detect differentiation specific genes, hormone dependent genes, and transcription-factor-induced genes. We conclude that DNA microarray is an excellent tool for identifying differentially expressed genes.

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The effect on gene expression profile of rat hippocampus caused by administration of memory enhancing herbal extract (육미지황탕가미방(六味地黃湯加味方)이 흰쥐의 기억능력과 중추신경계 유전자 발현에 미치는 영향)

  • Choi, Bo-Eop
    • Korean Journal of Oriental Medicine
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    • v.8 no.1
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    • pp.109-126
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    • 2002
  • The herbal extract (YMT_02) is a modified herbal extracts from Yukmijihwangtang (YMJ) to promote memory-enhancing. The YMJ extracts has been widely used as an anti-aging herbal medicine for hundred years in Asian countries. The purpose of this study is to; 1) quantitatively evaluate the memory-enhancing effect of YMT_02 by hehavior task, 2) identify candidate genes responsible for enhancing memory by cDNA microarray and 3) assess the anti-oxidant effect of YMT_02 on PC12 cell. Memory retention abilities are addressed by passive avoidance task with Sprague-Dawley (SD) male rat. Before the training session, the rats are subdivided into four groups and administrated with YMT_02, Ginkgo biloba, Soya lecithin and normal saline for 10 days. The retention test was performed. 24 hours after the training session. The retention time of the YMT_02 group was significantly (p<0.05) delayed $({\sim}100%)$, whereas Ginkgo biloba and Soya lecithin treatment delayed 20% and 10% respectively. The hippocampi of YMT_02 and control group were dissected and mRNA was further purified. After synthesizing cDNA using oligo-dT primer, the cDNA were applied and mRNA was further purified. After synthesizing cDNA using oligo-dT primer, the cDNA were applied to Incyte rat GEMTM 2 cDNA microarray. The microarray results show that prealbumin(transthyretin), phosphotidy lethanolamine N-methyltransferase, and PEP-19 are expressed abundantly in the YMT_02 treated group. Especially, PEP-19 is a neuron-specific protein, which inhibits apoptotic processes in neuronal cell. On the other hand, transcripts of RAB15, glutamate receptor subunit 2 and CDK 108 are abundant in control group. Besides, neuronal genes involved in neuronal death or neurodegeneration such as neuronal-pentraxin and spectrin are abundantly expressed in control group. Additionally, the YMT_02 shows an anti oxidative effect in the PC12 cell. The list of differentially expressed genes may implicate further insight on the action and mechanism behind the memory-enhancing effect of herbal extracts YMT_02, for example, anti-apoptotic, anti-oxidative, and neuroprotective effects.

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Acute Toxicity of Cadmium on Gene Expression Profiling of Fleshy Shrimp, Fenneropenaeus Chinensis Postlarvae Using a cDNA Microarray (Microarray 분석을 이용한 대하 (Fenneropenaeus chinensis) 유생의 카드뮴 단기 노출에 따른 유전자변화)

  • Kim, Su-Kyoung;Qiao, Guo;Yoon, Jong-Hwa;Jang, In-Kwon
    • Journal of Environmental Science International
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    • v.24 no.5
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    • pp.623-631
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    • 2015
  • Microarray technology provides a unique tool for the determination of gene expression at the level of messenger RNA (mRNA). This study, the mRNA expression profiles provide insight into the mechanism of action of cadmium in Fleshy shrimp (Fenneropenaeus chinensis). The ability of genomic technologies was contributed decisively to development of new molecular biomarkers and to the determination of new possible gene targets. Also, it can be approach for monitoring of trace metal using oligo-chip microarray-based in potential model marine user level organisms. 15K oligo-chip for F. chinensis that include mostly unique sets of genes from cDNA sequences was developed. A total of 13,971 spots (1,181 mRNAs up- regulated and 996 down regulated) were identified to be significantly expressed on microarray by hierarchical clustering of genes after exposure to cadmium for different conditions (Cd24-5000 and Cd48-1000). Most of the changes of mRNA expression were observed at the long time and low concentration exposure of Cd48-1000. But, gene ontology analysis (GO annotation) were no significant different between experiments groups. It was observed that mRNA expression of main genes involved in metabolism, cell component, molecular binding and catalytic function. It was suggested that cadmium inhibited metabolism and growth of F. chinensis.

Anti-inflammatory Effects of Extracts and Their Solvent Fractions of Rice Wine Lees (주박 추출물과 이들의 유기용매 분획물에 의한 항염증 활성)

  • Park, Mi-Jeong;Kang, Hyung-Taek;Kim, Mi-Sun;Shin, Woo-Chang;Sohn, Ho-Yong;Kim, Jong-Sik
    • Journal of Life Science
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    • v.24 no.8
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    • pp.843-850
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    • 2014
  • In the current study, we prepared eighty-five different kinds of solvent fractions of rice wine lees and nuruk extracts and investigated their effects on cell viability and nitric oxide (NO) production in mouse RAW 264.7 cells. Among the treated solvent fractions, only three solvent fractions (KSD-E1-3, KSD-E2-3 and KSD-E4-3) significantly decreased NO production in LPS-activated RAW 264.7 cells without affecting cell viability. And, they also reduced the expression of pro-inflammatory genes such as COX-2, TNF-alpha and iNOS. To understand the molecular mechanisms involved in the inhibition of inflammation in (KSD-E4-3)-treated RAW 264.7 cells, we carried out oligo DNA microarray analysis using Agilent Mouse microarray. To confirm microarray data, 6 genes (IL-1F6, iNOS, IL-10, Fabp4, IL-1RN and CSF2) were selected and performed RT-PCR and quantitative real-time PCR analysis with gene specific primers. The results of RT-PCR and real-time PCR agreed with microarray data. Overall, our results suggest that rice wine lees can be a novel resource for the development of foods and drugs which possess anti-inflammatory activity.

Quantitative analysis using decreasing amounts of genomic DNA to assess the performance of the oligo CGH microarray

  • Song Sunny;Lazar Vladimir;Witte Anniek De;Ilsley Diane
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2006.02a
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    • pp.71-76
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    • 2006
  • Comparative genomic hybridization (CGH) is a technique for studying chromosomal changes in cancer. As cancerous cells multiply, they can undergo dramatic chromosomal changes, including chromosome loss, duplication, and the translocation of DNA from one chromosome to another. Chromosome aberrations have previously been detected using optical imaging of whole chromosomes, a technique with limited sensitivity, resolution, quantification, and throughput. Efforts in recent years to use microarrays to overcome these limitations have been hampered by inadequate sensitivity, specificity and flexibility of the microarray systems. The oligonucleotide CGH microarray system overcomes several scientific hurdles that have impeded comparative genomic studies of cancer. This new system can reliably detect single copy deletions in chromosomes. The system includes a whole human genome microarray, reagents for sample preparation, an optimized microarray processing protocol, and software for data analysis and visualization. In this study, we determined the sensitivity, accuracy and reproducibility of the new system. Using this assay, we find that the performance of the complete system was maintained over a range of input genomic DNA from 5 ug down to 0.15 ug.

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DNA microarray analysis of RNAi plant regulated expression of NtROS2a gene encoding cytosine DNA demethylation (시토신 탈메틸화 관련 NtROS2a 유전자 발현을 제어한 RNAi 식물의 DNA microarray 분석)

  • Choi, Jang Sun;Lee, In Hye;Jung, Yu Jin;Kang, Kwon Kyoo
    • Journal of Plant Biotechnology
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    • v.43 no.2
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    • pp.231-239
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    • 2016
  • To study the transcript levels of epigenetically regulated genes in tobacco, we have developed a transgenic line OX1 overexpressing NtROS2a gene encoding cytosine DNA demethylation and a RNAi plant line RNAi13. It has been reported that salt- and $H_2O_2$-stress tolerance of these transgenic lines are enhanced with various phenotypic characters (Lee et al. 2015). In this paper, we conducted microarray analysis with Agilent Tobacco 4 x 44K oligo chip by using overexpression line OX1, RNAi plant line RNAi 13, and wild type plant WT. Differentially expressed genes (DEGs) related to metabolism, nutrient supply, and various stressed were up-regulated by approximately 1.5- to 80- fold. DEGs related to co-enzymes, metabolism, and methylation functional genes were down-regulated by approximately 0.03- to 0.7- fold. qRT-PCR analysis showed that the transcript levels of several candidate genes in OX1 and RNAi lines were significantly (p < 0.05) higher than those in WT, such as genes encoding KH domain-containing protein, MADS-box protein, and Zinc phosphodiesterase ELAC protein. On the other hand, several genes such as those encoding pentatricopeptide (PPR) repeat-containing protein, histone deacetylase HDAC3 protein, and protein kinase were decreased by approximately 0.4- to 1.0- fold. This study showed that NtROS2a gene encoding DNA glycosylase related to demethylation could regulate adaptive response of tobacco at transcriptional level.

Analysis of Differentially Expressed Genes by Allicin in Human Colorectal Cancer Cell (마늘성분 Allicin에 의해 차별적으로 발현되는 유전자군의 발현 분석)

  • Kim, Kyung-Ho;Kim, Min-Jung;Kim, Jong-Sik;Pyo, Suhk-Neung;Kim, Byung-Oh
    • Microbiology and Biotechnology Letters
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    • v.38 no.4
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    • pp.442-447
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    • 2010
  • We investigated anti-cancer and anti-proliferative activity of allicin and analyzed global gene expression changes by allicin treatment in human colorectal HCT116 cells. As a result, allicin decreased cell viabilities in a dose and time-dependent manner and induced apoptosis. Oligo DNA microarray analysis, we found that 7,840 genes were up-regulated more than 2-folds, whereas 10,010 genes were down-regulated more than 2-folds by $50\;{\mu}M$ allicin treatment. To confirm specific gene expressions, we performed RT-PCR. Consistent with the results of DNA microarray analysis, allicin dramatically induced ATF3 and NAG1 gene expression. Interestingly, NAG-1 protein expression was dependent on p53 presence. Taken together, our present results increase the knowledge of the molecular mechanism of anti-cancer and anti-proliferative activity mediated by allciin in human colorectal cancer cell.

Identification of Cuts-specific Myogenic Marker Genes in Hanwoo by DNA Microarray (DNA Microarray 분석을 통한 한우 부위별 특이 마커 유전자의 발굴)

  • Lee, Eun-Ju;Shin, Yu-Mi;Lee, Hyun-Jeong;Yoon, Du-Hak;Chun, Tae-Hoon;Lee, Yong-Seok;Choi, In-Ho
    • Journal of Animal Science and Technology
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    • v.52 no.4
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    • pp.329-336
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    • 2010
  • Myogenic satellite cells (MSCs) are mononuclear, multipotent progenitors of adult skeletal muscle possessing a capacity of forming adipocyte-like cells (ALC). To identify the skeletal muscle type-specific myogenic and adipogenic genes during MSCs differentiation, total RNA was extracted from bovine MSCs, myotube-formed cell (MFC), and ALC from each of Beef shank, Longissimus dorsi, Deep pectoral, and Semitendinosus. DNA microarray analysis (24,000 oligo chip) comparing MSCs with MFC and ALC, respectively, revealed 135 differentially expressed genes (> 4 fold) among four cuts. Real-time PCR confirmed expression of 29 genes. Furthermore, the whole tissue sample RNAs analysis showed 6 differentially expressed genes in Beef shank. Among which, 1 gene in MSCs, 4 in MFC, and 1 in ALCs were highly expressed. This study will provide an insight for better understanding the molecular mechanism of differentiation of skeletal muscle type-specific MSCs. The identified genes may be used as marker to distinguish skeletal muscle types.