• Title/Summary/Keyword: cDNA microarray analysis

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Construction and Analysis of a DNA Microarray for the Screening of Biosynthetic Genes of Secondary-Metabolites formation in Streptomyces (방선균 유래 이차대사 생합성 유전자 분석용 DNA Microarray 제작 및 해석)

  • Nam Soo Jung;Kang Dae-Kyung;Rhee Ki Hyeong;Kim Jong-Hee;Kang Sang Sun;Chang Yong Keun;Hong Soon-Kwang
    • Korean Journal of Microbiology
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    • v.41 no.2
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    • pp.105-111
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    • 2005
  • Streptomyces produces many kinds of secondary-metabolites including antibiotics. Screening of a new compound and elucidation of a biosynthetic pathway for the secondary metabolites are very important fields of biology, however, there is a main problem that most of the identified compounds are already researched compounds. To solve these problems, a microarray system that is based on the data related to the biosynthetic genes for secondary-metabolites was designed. For the main contents of DNA microarray, the important genes for the bio-synthesis of aminoglycosides, polyenes group, enediyne group, alpha-glucosidase inhibitors, glycopeptide group, and orthosomycin group were chosen. A DNA microarray with 69 genes that were involved in the bio-synthesis for the antibiotics mentioned above was prepared. The usability of the DNA microarray was confirmed with the chromosomal DNA and total RNA extracted from S. coelicolor whose genomic sequence had already been reported.

cDNA microarray analysis of viral hemorrhagic septicemia infected olive flounder, Paralichthys olivaceus: immune gene expression at different water temperature (바이러스성 출혈성 패혈증에 감염된 넙치의 cDNA microarray 분석 : 수온에 따른 면역 유전자 발현의 차이)

  • Kim, Jin-Ung;Jung, Sung-Ju
    • Journal of fish pathology
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    • v.27 no.1
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    • pp.1-9
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    • 2014
  • The olive flounder, Paralichthys olivaceus is susceptible to viral hemorrhagic septicaemia virus (VHSV) at $15^{\circ}C$ but no mortality at $20^{\circ}C$ even though the virus can grow well in vitro at $20^{\circ}C$. Thus, we designed an experiment to know immune response of olive flounder against VHSV when the host reared at $15^{\circ}C$ or $20^{\circ}C$. cDNA microarray analysis was performed to compare the gene expression patterns of the kidney cells between the host reared at $15^{\circ}C$ or $20^{\circ}C$. The expression of MHC class I, IL-8, myeloperoxidae and endonuclease G-like having function for the antigen presentation and chemokine-factor were up-regulted both the $15^{\circ}C$ and $20^{\circ}C$ during VHSV infection. MHC class II gene existing on antigen-presenting cells and B cell lymphocytes, immunoglobulin (Ig) genes and phagocytosis related genes were down-regulated at $15^{\circ}C$ but highly expressed at $20^{\circ}C$. It can be thought that innate immune related antigen presentation by MHC class I and phagocytosis reaction against VHSV are efficiently occur both the temperature but macrophage or B cell related antigen presentation via MHC class II fails to induce downstream immune reactions (adaptive immunity) to make antibody, and it can be one of the reason that causes high mortality only at $15^{\circ}C$.

DNA microarray analysis of gene expression of MC3T3-E1 osteoblast cell cultured on anodized- or machined titanium surface

  • Park, Ju-Mi;Jeon, Hye-Ran;Pang, Eun-Kyoung;Kim, Myung-Rae;Kang, Na-Ra
    • Journal of Periodontal and Implant Science
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    • v.38 no.sup2
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    • pp.299-308
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    • 2008
  • Purpose: The aim of this study was to evaluate adhesion and gene expression of the MC3T3-E1 cells cultured on machined titanium surface (MS) and anodized titanium surface (AS) using MTT test, Scanning electron micrograph and cDNA microarray. Materials and Methods: The MTT test assay was used for examining the proliferation of MC3T3-E1 cells, osteoblast like cells from Rat calvaria, on MS and AS for 24 hours and 48 hours. Cell cultures were incubated for 24 hours to evaluate the influence of the substrate geometry on both surfaces using a Scanning Electron Micrograph (SEM). The cDNA microarray Agilent Rat 22K chip was used to monitor expressions of genes. Results: After 24 hours of adhesion, the cell density on AS was higher than MS (p < 0.05). After 48 hours the cell density on both titanium surfaces were similar (p > 0.05). AS had the irregular, rough and porous surface texture. After 48 hours incubation of the MC3T3-E1 cells, connective tissue growth factor (CTGF) was up-regulated on AS than MS (more than 2 fold) and the insulin-like growth factor 1 receptor was down-regulated (more than 2 fold) on AS than MS. Conclusion: Microarray assay at 48 hours after culturing the cells on both surfaces revealed that osteoinductive molecules appeared more prominent on AS, whereas the adhesion molecules on the biomaterial were higher on MS than AS, which will affect the phenotype of the plated cells depending on the surface morphology.

Web-based microarray analysis using the virtual chip viewer and bioconductor. (MicroArray의 직관적 시각적 분석을 위한 웹 기반 분석 도구)

  • Lee, Seung-Won;Park, Jun-Hyung;Kim, Hyun-Jin;Kang, Byeong-Chul;Park, Hee-Kyung;Kim, In-Ju;Kim, Cheol-Min
    • Proceedings of the Korea Inteligent Information System Society Conference
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    • 2005.05a
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    • pp.198-201
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    • 2005
  • DNA microarray 칩은 신약 개발, 유전적 질환 진단, Bio-molecular 상호작용 연구, 유전자의 기능연구 등 폭넓게 사용되고 있다. 이 논문은 cDNA mimcroarray 데이터를 분석하기 위한 웹형태의 시스템 개발에 대한 내용을 다룬다. 하나의 cDNA microarray에는 수 백에서 수 만개의 유전자가 심어져 있으며, 데이터를 분석할 때 대량의 데이터와 다양한 형태의 오류로 인해서 데이터간의 차이를 보정하는 분석 도구와 통계적 기법들이 사용되어야 한다. 본 논문에서는 가상 칩 뷰어를 이용하여 실제 microarray 데이터의 foreground intensity에서 백그라운드의 intensity를 제거하여 일반화된 칩 이미지를 생성한다. 이 가상 칩 뷰어는 여러 가지 필터효과와 서로 다른 두 형광의 차이를 조정하는 global normalization 기법을 사용하여 발현 유전자 분석을 시각적으로 할 수 있고, 중복된 마이크로어레이 칩 데이터를 통하여 시간이 많이 걸리는 분석전 칩의 유효성을 검토할 수 있다. 칩 데이터의 normalization을 위한 통계 방법으로 R 통계 도구와 linear 모델을 사용하여 microarray 칩의 유전자 발현 양상을 분석한다. 통계적 방법을 사용하지 않은 데이터를 추출, 이 데이터의 패턴 그래프 그리고 발현 레벨을 분류하여 마이크로어레이의 각 스팟의 유효성 검토의 정확성을 높였다. 이 시스템은 칩의 유효성 검토, 스팟의 유효성 검토, 유전자 선정에 대해 분석의 용이성과 정확성을 높일 수 있었다.

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A Graph Model and Analysis Algorithm for cDNA Microarray Image (cDNA 마이크로어레이 이미지를 위한 그래프 모델과 분석 알고리즘)

  • Jung, Ho-Youl;Hwang, Mi-Nyeong;Yu, Young-Jung;Cho, Hwan-Gue
    • Journal of KIISE:Computer Systems and Theory
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    • v.29 no.7
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    • pp.411-421
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    • 2002
  • In this Paper we propose a new Image analysis algorithm for microarray processing and a method to locate the position of the grid cell using the topology of the grid spots. Microarray is a device which enables a parallel experiment of 10 to 100 thousands of test genes in order to measure the gene expression. Because of the huge data obtained by a experiment automated image analysis is needed. The final output of this microarray experiment is a set of 16-bit gray level image files which consist of grid-structured spots. In this paper we propose one algorithm which located the address of spots (spot indices) using graph structure from image data and a method which determines the precise location and shape of each spot by measuring the inclination of grid structure. Several experiments are given from real data sets.

Microarray Analysis of the Gene Expression Profiles of SL2 Cells Stimulated by LPS/PGN and Curdlan

  • Jin, Li Hua;Choi, Jung Kyoon;Cho, Hwan Sung;Shim, Jaewon;Kim, Young-Joon
    • Molecules and Cells
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    • v.25 no.4
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    • pp.553-558
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    • 2008
  • Essential aspects of the innate immune response to microbial infection appear to be conserved between insects and mammals. In order to identify new Drosophila melanogaster genes involved in the immune response, we performed gene expression profiling of Drosophila SL2 cells stimulated with bacterial (LPS/PGN) or fungal (curdlan) components using a cDNA microarray that contained 5,405 Drosophila cDNAs. We found that some genes were similarly regulated by LPS/PGN and curdlan. However, a large number, belonging to the functional classes of cell organization, development, signal transduction, morphogenesis, cell cycle, and DNA replication, displayed significant differences in their transcription profiles between the two treatments, demonstrating that bacterial and fungal components induce different immune response even in an in vitro cell system.

Microarray Analysis of Differential Gene Expression in Alcohol-Exposed Post Implantation Embryos

  • Kim, So-Hee;Rhee, Gyu-Seek;Kim, Soon-Sun;Sohn, Kyung-Hee;Kwack, Seung-Jun;Lee, Rhee-Da;Chae, Soo-Yeong;An, Sang-Mi;Hur, Man-Wook
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2002.11b
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    • pp.204-204
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    • 2002
  • Alcohol drinking during pregnancy can result in abnormal fetal development including fetal alcohol syndrome (FAS). The molecular mechanisms of FAS, however, is not completely elucidated. In the present study, we evaluated the developmental toxicity of ethanol and its metabolite, acetaldehyde using post implantation whole embryo culture and determined changes of gene expression by ethanol treatment by cDNA microarray.(omitted)

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Global Analysis of Estrogen-Regulated Genes in Mouse Uterus using cDNA Microarray and Laser Capture Microdissection (cDNA Microarray와 Laser Capture Microdissection을 이용한 생쥐 자궁에서 Estrogen에 의해 조절되는 유전자 발현에 관한 분석)

  • Hong, Seok-Ho;Nah, Hee-Young;Lee, Ji-Yoon;Kim, Chung-Hoon;Kim, Moon-Kyoo
    • Clinical and Experimental Reproductive Medicine
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    • v.30 no.2
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    • pp.151-163
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    • 2003
  • 연구목적: Estrogen은 포유류의 생리주기와 착상과정에서 중요한 조절인자로 작용한다. 본 연구에서는 난소 절제된 생쥐의 자궁에서 estrogen에 의해 직접 또는 간접적으로 조절되어 발현하는 유전자를 분석하고자 하였다. 연구재료 및 방법: 생후 8주된 생쥐의 양쪽 난소를 절제하고 14일 동안 회복기간이 지난 후, estrogen (300 ng/mouse)을 피하로 주사하였다. Estrogen 주사 후 6, 12시간째 자궁을 적출하여 cDNA microarray와 laser capture microdissection (LCM) 기술을 이용하여 estrogen에 의해 조절되는 유전자의 시공간적인 발현 양상을 조사하였다. 결 과: Estrogen 주사 후 6시간째에는 조사된 전체 유전자 가운데 0.9% (증가 22, 감소 49), 12시간째에는 8.4% (증가 351, 감소 287)에 해당되는 유전자가 두 배 이상 증가 혹은 감소하는 결과를 보였다. 또한 일부 증감된 유전자를 선택한 후 LCM 기술을 이용하여 시공간적인 발현양상을 확인한 결과 자궁내막상피세포에서만 estrogen에 의해 유전자의 발현이 증가되는 일부 유전자를 선별하였다. 결 론: 이상의 결과들을 종합해보면 1) estrogen에 의해 조절되는 유전자의 수나 증감의 정도는 12시간 이후에 더 많고, 크게 조절되며, 2) 유전자의 조절부위가 자궁의 특이적인 세포층에서 시공간적으로 조절됨을 의미한다. 이러한 유전자의 정보는 생리주기 또는 착상과정의 분자생물학적 기작을 이해하는 데 도움이 될 것이다.