• Title/Summary/Keyword: cDNA Microarray

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Molecular Cloning of Adipose Tissue-specific Genes by cDNA Microarray

  • Kim, Kee-Hong;Moon, Yang Soo
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.12
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    • pp.1837-1841
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    • 2003
  • In an attempt to isolate novel molecules that may play a regulatory role in adipocyte differentiation, we devised an experimental strategy to identify adipose tissue-specific genes by modifying cDNA microarray technique. We used genefilter membranes containing approximately 15,000 rat non-redundant EST clones of which 4,000 EST were representative clones of known genes and 11,000 ESTs were uncharacterized clones. A series of hybridization of genefilter membranes with cDNA probes prepared from various rat tissues and nucleic acids sequence analysis allowed us to identify two adipose-tissue specific genes, adipocyte-specific secretory factor (ADSF) and H-rev107. Verification of tissue-specific expression patterns of these two genes by Northern blot analysis showed that ADSF mRNA is exclusive expressed in adipose tissue and the H-rev107 mRNA is predominantly expressed in adipose tissue. Further analysis of gene expression of ADSF and H-rev107 during 3T3-L1 adipocyte differentiation revealed that the ADSF and H-rev107 gene expression patterns are closely associated with the adipocyte differentiation program, indicating their possible role in the regulation of adipose tissue development. Overall, we demonstrated an application of modified cDNA microarray technique in molecular cloning, resulting in identification of two novel adipose tissue-specific genes. This technique will also be used as a useful tool in identifying novel genes expressed in a tissue-specific manner.

An Iterative Normalization Algorithm for cDNA Microarray Medical Data Analysis

  • Kim, Yoonhee;Park, Woong-Yang;Kim, Ho
    • Genomics & Informatics
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    • v.2 no.2
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    • pp.92-98
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    • 2004
  • A cDNA microarray experiment is one of the most useful high-throughput experiments in medical informatics for monitoring gene expression levels. Statistical analysis with a cDNA microarray medical data requires a normalization procedure to reduce the systematic errors that are impossible to control by the experimental conditions. Despite the variety of normalization methods, this. paper suggests a more general and synthetic normalization algorithm with a control gene set based on previous studies of normalization. Iterative normalization method was used to select and include a new control gene set among the whole genes iteratively at every step of the normalization calculation initiated with the housekeeping genes. The objective of this iterative normalization was to maintain the pattern of the original data and to keep the gene expression levels stable. Spatial plots, M&A (ratio and average values of the intensity) plots and box plots showed a convergence to zero of the mean across all genes graphically after applying our iterative normalization. The practicability of the algorithm was demonstrated by applying our method to the data for the human photo aging study.

Genes expression by using cDNA Microarray in Whallak-tang (활락탕(活絡湯)의 cDNA Microarray를 이용한 유전자 발현에 미치는 영향)

  • Sin, Cheol-Kyung;Lee, Chae-Woo;Yoo, Sun-Ae;Youn, Hyoun-Min;Jang, Kyung-Jeon;Song, Choon-Ho;Ahn, Chang-Beohm;Kim, Cheol-Hong
    • Journal of Pharmacopuncture
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    • v.11 no.4
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    • pp.5-14
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    • 2008
  • Objective This study was undertaken to determine the effect of Whallak-tang on expression of CD/cytokine Genes. Methods The expression of CD/Cytokine Genes were examined by cDNA microarray using the human mast cell line(HMC-1). Results The expression of ATP5F1, FLJ20671, unknown, KIAA0342, OAS2, unknown genes were increased in $200{\sim}300%$ range. The expression of unknown, MDS006, IFITM1, MRPL3, ZNF207, FTH1, FBP1, NRGN, NR1H2, KIAA0747 genes were decreased in $0{\sim}33%$ range. Conclusion These results would provide important basic data on the possibility of the clinical treatment of Whallak-tang in musculoskeletal disease.

Survey of Genes Responsive to Long-Term Heat Stress Using a cDNA Microarray Analysis in Mud Loach (Misgurnus mizolepis) Liver (장기 고온 스트레스에 대한 미꾸라지(Misgurnus mizolepis) 간 조직 내 유전자 발현 반응의 cDNA microarray 분석)

  • Cho, Young Sun;Lee, Sang Yoon;Noh, Choong Hwan;Nam, Yoon Kwon;Kim, Dong Soo
    • Korean Journal of Ichthyology
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    • v.18 no.2
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    • pp.65-77
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    • 2006
  • Gene transcripts potentially responsive to the heat stress were surveyed by cDNA microarray analysis in mud loach (Misgurnus mizolepis). Transcriptional profiles of hepatic tissue in the fish exposed to either $23^{\circ}C$ or $32^{\circ}C$ for 4 weeks were compared each other by 3 replicated hybridization assays using 1,124 unigene clones selected from mud loach liver expressed sequence tags (ESTs). A total of 93 clones showed the substantially increased mRNA levels (>2-fold) in $32^{\circ}C$-exposed group when compared in $23^{\circ}C$control group. It includes various enzymes and proteins involved in energy pathway, protease/protein metabolisms, immune/antioxidant functions, cytoskeleton/cell structure, transport and/or signal transduction. Maximum level of increase was up to 15-fold relative to $23^{\circ}C$ treatment. Heat exposure also resulted in the significant decrease (less than 50% relative to $23^{\circ}C$-exposed fish) of the transcriptional activities in 85 genes. Besides the above categories, yolk protein (vitellogenin) and ribosomal proteins were notably down regulated in the fish exposed to heat stress. A number of novel gene transcripts were also detected in both up-regulated and down-regulated groups.

Patterns of Intrahepatic Gene Expression in Neonatal Cholestasis (신생아 담즙정체성 간질환에서 간조직 유전자의 발현 양상)

  • Choi, BoHwa;Choe, Byung Ho;Chung, Eun Jung;Kim, Kyung Mo;Kim, Heng Mi;Park, Jin Young;Park, Woo Hyun;Kim, Moon Kyu;Kim, Jung Chul
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.8 no.2
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    • pp.177-193
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    • 2005
  • Purpose: To identify genes specifically expressed in biliary atresia, we compared the patterns of gene expression between biliary atresia and neonatal hepatitis syndrome using cDNA microarray analysis. Methods: Liver tissues were taken from livers of 11 patients (7 patients with biliary atresia and four with neonatal hepatitis) with neonatal cholestasis by needle biopsy. Normal control could be obtained from donor liver tissue during living-related liver transplantation. Total RNA was extracted from each samples and reversely transcribed to make cDNA. Then fluorescent cDNA were pooled and hybridized to the clones on the microarray. Fluorescence intensities at the immobilized targets were measured. Utilizing cDNA arrays of 4.7 K human genes, gene expression profiles were analyzed. Results: Among 4,700 microarray clones, 17 cDNA clones were significantly over-expressed in all 11 patients with neonatal cholestasis, while 20 clones were significantly decreased. Genome-wide expression analysis was carried out in livers obtained at the time of diagnosis. We could identify 49 genes, in which there showed differential expression between biliary atresia and neonatal hepatitis syndrome. Conclusion: This study shows the pattern of differentially expressed genes in biliary atresia and neonatal hepatitis syndrome. We believe that this study can contribute to the understanding of pathogenesis of neonatal cholestasis.

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Statistical Methods for Gene Expression Data

  • Kim, Choongrak
    • Communications for Statistical Applications and Methods
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    • v.11 no.1
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    • pp.59-77
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    • 2004
  • Since the introduction of DNA microarray, a revolutionary high through-put biological technology, a lot of papers have been published to deal with the analyses of the gene expression data from the microarray. In this paper we review most papers relevant to the cDNA microarray data, classify them in statistical methods' point of view, and present some statistical methods deserving consideration and future study.

Print-tip Normalization for DNA Microarray Data (DNA 마이크로어레이 자료의 PRINT-TIP별 표준화(NORMALIZATION) 방법)

  • Yi Sung-Gon;Park Taesung;Kang Sung Hyun;Lee Seung-Yeaun;Lee Yang Sung
    • The Korean Journal of Applied Statistics
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    • v.18 no.1
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    • pp.115-127
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    • 2005
  • DNA microarray experiments allow us to study expression of thousands of genes simultaneously, Normalization is a process for removing noises occurred during the microarray experiment, Print-tip is regarded as one main sources of noises, In this paper, we review normalization methods most commonly used in the microarray experiments, Especially, we investigate the effects of print-tips through simulated data sets.

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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Platform of Hot Pepper Stress Genomics: Indentification of Stress Inducible Genes in Hot Pepper (Capsicum annuum L.) Using cDNA Microarray Analysis

  • Chung, Eun-Jo;Lee, Sanghyeob;Park, Doil
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.81.1-81
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    • 2003
  • Although plants have evolved to possess various defense mechanisms from local biotic and abiotic stressors, most of yield loss is caused by theses stressors. Recent studies have revealed that several different stress responsive reactions are inter-networking. Therefore, the identification and dissection of stress responsive genes is an essential and first step towards understanding of the global defense mechanism in response to various stressors. For this purpose, we applied cDNA microarray analysis, because it has powerful ability to monitor the global gene expression in a specific situation. To date, more than 10,000 non-redundant genes were identified from seven different cDNA libraries and deposited in our EST database (http://plant.pdrs.re.kr/ks200201/pepper.html). For this study, we have built 5K cDNA microarray containing 4,685 unigene clones from three different cDNA libraries. Monitoring of gene expression profiles of hot pepper interactions with biotic stress, abiotic stresses and chemical treatments will be presented. Although this work shows expression profiling at the sub-genomic level, this could be a good starting point to understand the complexity of global defense mechanism in hot pepper.

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Gene Expression Profile of Zinc-Deficient, Homocysteine-Treated Endothelial Cells

  • Kwun, In-Sook;Beattie, John H.
    • Preventive Nutrition and Food Science
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    • v.8 no.4
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    • pp.390-394
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    • 2003
  • In the post-genome period, the technique for identifying gene expression has been progressed to high throughput screening. In the field of molecular nutrition, the use of screening techniques to clarify molecular function of specific nutrients would be very advantageous. In this study, we have evaluated Zn-regulated gene expression in Zn-deficient, homocystein-treated EA.hy926 cells, using cDNA microarray, which can be used to screen the expression of many genes simultaneously. The information obtained can be used for preliminary assessment of molecular and signaling events modulated by Zn under pro-atherogenic conditions. EA.hy926 cells derived from human umbilical vein endothelial cells were cultured in Zn-adequate (control, 15 $\mu$M Zn) or Zn-deficient (experimental, 0 $\mu$M Zn) Dulbecco's MEM media under high homocysteine level (100 $\mu$M) for 3 days of post-confluency. Cells were harvested and RNA was extracted. Total RNA was reverse-transcribed and the synthesized cDNA was labeled with Cy3 or Cy5. Fluorescent labeled cDNA probe was applied to microarray slides for hybridization, and the slide was then scanned using a fluorescence scanner. The expression of seven genes was found to be significantly decreased, and one significantly increased, in response to treatment of EA.hy926 cells with Zn-deficient medium, compared with Zn-supplemented medium. The upregulated genes were oncogenes and tumor suppressor genes, cell cycle-related genes and transporter genes. The down-regulated gene was RelB, a component of the NF-kappaB complex of transcription factors. The results of this study imply the effectiveness of cDNA microarray for expression profiling of a singly nutrient deficiency, namely Zn. Furthur study, using tailored-cDNA array and vascular endothelial cell lines, would be beneficial to clarify the molecular function of Zn in atherosclerosis, more in detail.