• 제목/요약/키워드: cDNA Microarray Analysis

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The Study of X Chromosome Inactivation Mechanism in Klinefelter's Syndrome by cDNA Microarray Experiment

  • Jeong, Yu-Mi;Chung, In-Hyuk;Park, Jung Hoon;Lee, Sook-Hwan;Chung, Tae-Gyu;Kim, Yong Sung;Kim, Nam-Soon;Yoo, Hyang-Sook;Lee, Suman
    • Genomics & Informatics
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    • v.2 no.1
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    • pp.30-35
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    • 2004
  • To investigate the XIST gene expression and its effect in a Klinefelter's patient, we used Klinefelter's syndrome (XXY) patient with azoospermia and also used a normal male (XY) and a normal female (XX) as the control, We were performed cytogenetic analysis, Y chromosomal microdeletion assay (Yq), semi-quantitative RT-PCR, and the Northern blot for Klinefelter's syndrome (KS) patient, a female and a male control, We extracted total RNA from the KS patient, and from the normal cells of the female and male control subjects using the RNA prep kit (Qiagen), cDNA microarray contained 218 human X chromosome-specific genes was fabricated. Each total RNA was reverse transcribed to the first strand cDNA and was labeled with Cy-3 and Cy-5 fluorescein, The microarray was scanned by ScanArray 4000XL system. XIST transcripts were detected from the Klinefelters patient and the female by RT-PCR and Northern blot analysis, but not from the normal male, In the cDNA microarray experiment, we found 24 genes and 14 genes are highly expressed in KS more than the normal male and females, respectively. We concluded that highly expressed genes in KS may be a resulted of the abnormal X inactivation mechanism.

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 ANALYSIS OF THE DENTAL PULP IN HEALTHY AND CARIES TEETH (치아 우식증에 따른 치수내 유전자 발현 변화에 관한 분석)

  • Oh, So-Hee;Kim, Jong-Soo
    • Journal of the korean academy of Pediatric Dentistry
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    • v.37 no.3
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    • pp.275-287
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    • 2010
  • Deep caries may induce pulpitis and the pulpal tissue interacts with microbial invasion. The immune response to protect the pulpal tissue can be mediated by cellular signal molecules produced by the pulpal cells. The understanding of these processes is important to find future therapeutic method for the diseased pulp. The pulp tissue from sound teeth was set as control group (n=30) and the pulp tissue from decayed teeth was set as test group (n=30). Total RNA was extracted from the pulp of each group and it was used for cDNA microarray and reverse transcriptase-polymerase chain reaction(RT-PCR). The expression of TGF-${\beta}1$ was studied by immunohistochemistry. The results were as follows: 1. cDNA microarray analysis identified 520 genes with 6-fold or greater difference in expression level with 143 genes more abundant in health and 377 genes more abundant in disease. 2. The RT-PCR analysis was done for randomly selected 14 genes and the results supported the result of cDNA microarray assay. 3. TGF-${\beta}1$ was highly expressed in the carious pulp and it was found in odontoblast by immunohistochemistry. In conclusion, many cytokines were found to be significantly changed their expression in the diseased pulp(/M/>1.6).

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.

Two-Stage Logistic Regression for Cancer Classi cation and Prediction from Copy-Numbe Changes in cDNA Microarray-Based Comparative Genomic Hybridization

  • Kim, Mi-Jung
    • The Korean Journal of Applied Statistics
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    • v.24 no.5
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    • pp.847-859
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    • 2011
  • cDNA microarray-based comparative genomic hybridization(CGH) data includes low-intensity spots and thus a statistical strategy is needed to detect subtle differences between different cancer classes. In this study, genes displaying a high frequency of alteration in one of the different classes were selected among the pre-selected genes that show relatively large variations between genes compared to total variations. Utilizing copy-number changes of the selected genes, this study suggests a statistical approach to predict patients' classes with increased performance by pre-classifying patients with similar genetic alteration scores. Two-stage logistic regression model(TLRM) was suggested to pre-classify homogeneous patients and predict patients' classes for cancer prediction; a decision tree(DT) was combined with logistic regression on the set of informative genes. TLRM was constructed in cDNA microarray-based CGH data from the Cancer Metastasis Research Center(CMRC) at Yonsei University; it predicted the patients' clinical diagnoses with perfect matches (except for one patient among the high-risk and low-risk classified patients where the performance of predictions is critical due to the high sensitivity and specificity requirements for clinical treatments. Accuracy validated by leave-one-out cross-validation(LOOCV) was 83.3% while other classification methods of CART and DT performed as comparisons showed worse performances than TLRM.

Radioactive cDNA microarray in Neurospsychiatry (신경정신 의학분야의 방사성동위원소 표지 cDNA 마이크로어레이)

  • Choe, Jae-Gol;Shin, Kyung-Ho;Lee, Min-Soo;Kim, Meyoung-Kon
    • The Korean Journal of Nuclear Medicine
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    • v.37 no.1
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    • pp.43-52
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    • 2003
  • Microarray technology allows the simultaneous analysis of gene expression patterns of thousands of genes, in a systematic fashion, under a similar set of experimental conditions, thus making the data highly comparable. In some cases arrays are used simply as a primary screen loading to downstream molecular characterization of individual gene candidates. In other cases, the goal of expression profiling is to begin to identify complex regulatory networks underlying developmental processes and disease states. Microarrays were originally used with ceil lines or other simple model systems. More recently, microarrays have been used in the analysis of more complex biological tissues including neural systems and the brain. The application of cDNA arrays in neuropsychiatry has lagged behind other fields for a number of reasons. These include a requirement for a large amount of input probe RNA In fluorescent-glass based array systems and the cellular complexity introduced by multicellular brain and neural tissues. An additional factor that impacts the general use of microarrays in neuropsychiatry is the lack of availability of sequenced clone sets from model systems. While human cDNA clones have been widely available, high qualify rat, mouse, and drosophilae, among others are just becoming widely available. A final factor in the application of cDNA microarrays in neuropsychiatry is cost of commercial arrays. As academic microarray facilitates become more commonplace custom made arrays will become more widely available at a lower cost allowing more widespread applications. in summary, microarray technology is rapidly having an impact on many areas of biomedical research. Radioisotope-nylon based microarrays offer alternatives that may in some cases be more sensitive, flexible, inexpensive, and universal as compared to other array formats, such as fluorescent-glass arrays. In some situations of limited RNA or exotic species, radioactive membrane microarrays may be the most practical experimental approach in studying psychiatric and neurodegenerative disorders, and other complex questions in the brain.

Comparison of Normalizations for cDNA Microarray Data

  • Kim, Yun-Hui;Kim, Ho;Park, Ung-Yang;Seo, Jin-Yeong;Jeong, Jin-Ho
    • Proceedings of the Korean Statistical Society Conference
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    • 2002.05a
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    • pp.175-181
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    • 2002
  • cDNA microarray experiments permit us to investigate the expression levels of thousands of genes simultaneously and to make it easy to compare gene expression from different populations. However, researchers are asked to be cautious in interpreting the results because of the unexpected sources of variation such as systematic errors from the microarrayer and the difference of cDNA dye intensity. And the scanner itself calculates both of mean and median of the signal and background pixels, so it follows a selection which raw data will be used in analysis. In this paper, we compare the results in each case of using mean and median from the raw data and normalization methods in reducing the systematic errors with arm's skin cells of old and young males. Using median is preferable to mean because the distribution of the test statistic (t-statistic) from the median is more close to normal distribution than that from mean. Scaled print tip normalization is better than global or lowess normalization due to the distribution of the test-statistic.

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Nonlinear matching measure for the analysis of on-off type microarray image (온-오프 형태의 DNA 마이크로어레이 영상 분석을 위한 비선형 정합도)

  • Ryu Mun ho;Kim Jong dae
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.30 no.3C
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    • pp.112-118
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    • 2005
  • In this paper, we propose a new nonlinear matching measure for automatic analysis of the on-off type DNA microarray images in which the hybridized spots are detected by the template matching method. The proposed measure is obtained by binary-thresholding over the whole template region and taking the number of white pixels inside the spotted area. This measure is compared with the normalized covariance in terms of the classification ability of the successfulness of the locating markers. The proposed measure is evaluated for the scanned images of HPV DNA microarrays where the marker locating is a critical issue because of the small number of spots. The targeting spots of HPV DNA chips are designed for genotyping 22 types of the human papilloma virus(HPV). The proposed measure is proven to give more discriminative response reducing the miss cases of the successful marker locating.

Profile of Gene Expression Changes During Doxorubicin Induced Apoptosis of Saos-2 (Saos-2 세포에서 Doxorubicin에 의한 세포사멸 유도과정에서의 유전자 발현 변화)

  • Lim, Jeong-Sook;Bae, Min-Jae;Baek, Suk-Hwan;Kim, Jae-Ryong;Kim, Jung-Hye;Kim, Seong-Yong
    • Journal of Yeungnam Medical Science
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    • v.22 no.2
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    • pp.221-240
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    • 2005
  • Background: Doxorubicin has proved to be a useful chemotherapeutic agent especially for osteogenic sarcoma. It induces cancer cell death via apoptosis. Materials and Methods: To explore and analyze the changes of gene expression during doxorubicin induced apoptosis on human osteogenic sarcoma, Saos-2 cell, cDNA microarray was performed. After treatment with doxorubicin, total RNA was purified and expressed genes were investigated with a 17k human cDNA microarray. Results: For analysis of the cDNA microarray, the genes were filtered using the sum of the median value of Cy3 and Cy5 signal intensity of greater than 800. Expression of 264 genes was changed by more than 2 fold, and the expression of 35 genes was changed more than 3 fold after treatment with doxorubicin. The genes were primarily related to cell death, cell growth and maintenance, signal transduction, cellular component, transport, and metabolism. Conclusion: Treatment with doxorubicin induced expressional change of many genes. Some of the genes might be related with apoptosis directly or indirectly. Further study is now needed to characterize these genes.

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Statistical Analysis of a Loop Designed Microarray Experiment Data (되돌림설계를 이용한 마이크로어레이 실험 자료의 분석)

  • 이선호
    • The Korean Journal of Applied Statistics
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    • v.17 no.3
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    • pp.419-430
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    • 2004
  • Since cDNA microarray experiments can monitor expression levels for thousands of genes simultaneously, the experimental designs and their analyzing methods are very important for successful analysis of microarray data. The loop design is discussed for selecting differentially expressed genes among several treatments and the analysis of variance method is introduced to normalize microarray data and provide estimates of the interesting quantities. MA-ANOVA is used to illustrate this method on a recently collected loop designed microarray data at Cancer Metastasis Research Center, Yonsei University.