• Title/Summary/Keyword: Gene profiling

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DNAchip as a Tool for Clinical Diagnostics (진단의학 도구로서의 DNA칩)

  • 김철민;박희경
    • Proceedings of the Korean Institute of Intelligent Systems Conference
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    • 2004.04a
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    • pp.97-100
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    • 2004
  • The identification of the DNA structure as a double-stranded helix consting of two nucleotide chain molecules was a milestone in modern molecular biology. The DNA chip technology is based on reverse hybridization that follows the principle of complementary binding of double-stranded DNA. DNA chip can be described as the deposition of defined nucleic acid sequences, probes, on a solid substrate to form a regular array of elements that are available for hybridization to complementary nucleic acids, targets. DNA chips based on cDNA clons, oligonucleotides and genomic clons have been developed for gene expression studies, genetic variation analysis and genomic changes associated with disease including cancers and genetic diseases. DNA chips for gene expression profiling can be used for functional analysis in human eel Is and animal models, disease-related gene studies, assessment of gene therapy, assessment of genetically modified food, and research for drug discovery. DNA chips for genetic variation detection can be used for the detection of mutations or chromosomal abnormalities in cnacers, drug resistances in cancer cells or pathogenic microbes, histocompatibility analysis for transplantation, individual identification for forensic medicine, and detection and discrimination of pathogenic microbes. The DNA chip will be generalized as a useful tool in clinical diagnostics in near future. Lab-on-a chip and informatics will facilitate the development of a variety of DNA chips for diagnostic purpose.

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Gene Expression Analysis of Megakaryocytes Derived from Human Umbilical Cord $CD34^+$ Cells by Thrombopoietin

  • Kim, Jeong-Ah;Kim, Hyung-Lae
    • Genomics & Informatics
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    • v.3 no.1
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    • pp.8-14
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    • 2005
  • Although much is known about the molecular biology of platelets, the megakaryocytes' (MKs) molecular biology was not understood so well because of their rareness. By the cloning and characterization of thrombopoietin (TPO), which is the principal regulator of the growth and development of the MKs, researches on the MKs have been growing rapidly. To understand megakaryocytopoiesis, we investigated the gene expression profile of the MKs using oligonucleotide microarray where 10,108 unique genes were spotted. Comparing the fluorescence intensities of which ratio is $\ge$ ${\mid}2{\mid}$, 372 genes were up-regulated and 541 genes were down-regulated in MKs. For confirmatory expression, RNase protection assay (RPA) establishing abundant apoptotic gene expression was carried out. In MKs, many of the known genes, including several platelet related genes, GATA binding protein were highly expressed. Particularly, TGF beta, clusterin (complement lysis inhibitor), and thymosin beta 4 (actin-sequestering molecules) were expressed highly in MKs. As MKs specific expressed genes may regulate normal and pathologic platelet (and/or MK) functions, the transcript profiling using microarray was useful on molecular understanding of MKs,

Production of taxadiene from cultured ginseng roots transformed with taxadiene synthase gene

  • Cha, Mi-Jeong;Shim, Sang-Hee;Kim, Sung-Hong;Kim, Ok-Tae;Lee, Se-Weon;Kwon, Suk-Yoon;Baek, Kwang-Hyun
    • BMB Reports
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    • v.45 no.10
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    • pp.589-594
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    • 2012
  • Paclitaxel is produced by various species of yew trees and has been extensively used to treat tumors. In our research, a taxadiene synthase (TS) gene from Taxus brevifolia was used to transform the roots of cultured ginseng (Panax ginseng C.A. Meyer) to produce taxadiene, the unique skeletal precursor to taxol. The TS gene was successfully introduced into the ginseng genome, and the de novo formation of taxadiene was identified by mass spectroscopy profiling. Without any change in phenotypes or growth difference in a TS-transgenic ginseng line, the transgenic TSS3-2 line accumulated $9.1{\mu}g$ taxadiene per gram of dry weight. In response to the treatment of methyl jasmonate for 3 or 6 days, the accumulation was 14.6 and $15.9{\mu}g$ per g of dry weight, respectively. This is the first report of the production of taxadiene by engineering ginseng roots with a taxadiene synthase gene.

Gene Expression Profiling Reveals that Paeoniflorin Has an Apoptotic Potential in Human Leukemia U937 Cells

  • Lim, Soo-Hyun;Ahn, Kwang-Seok;Kim, Sung-Hoon;Jang, Hyeung-Jin
    • Molecular & Cellular Toxicology
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    • v.5 no.4
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    • pp.335-345
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    • 2009
  • A major source of paeoniflorin (PF) which was from the Paeonia lactiflora root, has been used as a herbal medicine in East Asia for its antiallergic, antiinflammatory, and immunoregulatory effects. However, only few details are known about the mechanism of apoptosis induced by this compound. The present study was undertaken to further elucidate the molecular mechanism of apoptosis and the changes of gene expression elicited by PF using DNA microarrays and computational gene-expression analysis tools in human leukemia U937 cells. A comparative global transcription analysis between treatment with PF and anisomycin (AM) that induces apoptosis in U937 cells revealed that c-Jun-$NH_2$-kinase (JNK) pathway related genes were less expressed in PF-treated cells. Elucidation of the mechanisms by which PF conducts its anti-cancer activities through comparative analysis of the gene expression is necessary to provide a solid foundation for its use as a promising agent in prevention and treatment strategies.

Gene Expression Analysis of Hepatic Response Induced by Gentamicin in Mice

  • Oh, Jung-Hwa;Park, Han-Jin;Hwang, Ji-Yoon;Jeong, Sun-Young;Lim, Jung-Sun;Kim, Yong-Bum;Yoon, Seok-Joo
    • Molecular & Cellular Toxicology
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    • v.3 no.1
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    • pp.60-67
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    • 2007
  • Gentamicin is a broad-spectrum aminoglycoside antibiotic used in the treatment of bacterial infection. Although side effects of gentamicin such as nephrotoxicity and ototoxicity have been investigated, the information on the hepatic effects of gentamicin is still limited. In the present study, gene expression profiles were analyzed in the liver of gentamicin treated mice using Affymetrix GeneChip$^{(R)}$ Mouse Expression 430A 2.0 Array. Totally, 400 genes were identified as being either up- or down-regulated over 1.5-fold changes (P<0.01) in the liver of gentamicin treated mice. Among these deregulated genes, 16 up-regulated genes mainly involved in transport (Kif5b, Pex14, Rab14, Clcn3, and Necap1) and 20 down-regulated genes involved in lipid and other metabolisms (Hdlbp, Gm2a, Uroc1, and Dak) were selected using k-means clustering algorithm. The functional classification of differentially expressed genes represented that several stress-related genes were regulated in the liver by gentamicin treatment. This data may contribute in understanding the molecular mechanism in the liver of gentamicin treated mice.

Characterization and Genetic Profiling of the Primary Cells and Tissues from Mandible of Mouse Fetus and Neonate

  • Kang, Jung-Han;Nam, Hyun;Park, Soon-Jung;Oh, Keun-Hee;Lee, Dong-Seup;Cho, Jae-Jin;Lee, Gene
    • International Journal of Oral Biology
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    • v.32 no.1
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    • pp.13-22
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    • 2007
  • The stem cell research is emerging as a cutting edge topic for a new treatment for many chronic diseases. Recently, dental stem cell would be possible for regeneration of tooth itself as well as periodontal tissue. However, the study of the cell characterization is scarce. Therefore, we performed the genetic profiling and the characterization of mouse fetus/neonate derived dental tissue and cell to find the identification during dental development. We separated dental arch from mandibles of 14.5 d fetal mice and neonate 0 d under the stereoscope, and isolated dental cells primarily from the tissues. Then, we examined morphology and the gene expression profiles of the primary cells and dental tissues from fetus/neonate and adult with RT-PCR. Primary dental cells showed heterogeneous but the majority was shown as fibroblast-like morphology. The change of population doubling time levels (PDLs) showed that the primary dental cells have growth potential and could be expanded under our culture conditions without reduction of growth rate. Immunocytochemical and flow cytometric analyses were performed to characterize the primary dental cell populations from both of fetus (E14.5) and neonate. Alpha smooth muscle actin (${\alpha}-SMA$), vimentin, and von Willebrand factor showed strong expression, but desmin positive cells were not detected in the primary dental cells. Most of the markers were not uniformly expressed, but found in subsets of cells, indicating that the primary dental cell population is heterogeneous, and characteristics of the populations were changed during culture period. And mesenchymal stem cell markers were highly expressed. Gene expression profile showed Wnt family and its related signaling molecules, growth factors, transcription factors and tooth specific molecules were expressed both fetal and neonatal tissue. The tooth specific genes (enamelin, amelogenin, and DSPP) only expressed in neonate and adult stage. These expression patterns appeared same as primary fetal and neonatal cells. In this study we isolated primary cells from whole mandible of fetal and neonatal mice. And we investigated the characteristics of the primary cells and the profile of gene expressions, which are involved in epithelial-mesenchymal interactions during tooth development. Taken together, the primary dental cells in early passages or fetal and neonatal mandibles could be useful stem cell resources.

Lipogenesis Gene Expression Profiling in Longissimus dorsi on the Early and Late Fattening stage of Hanwoo (한우 비육 전·후기의 등심조직에 있어서 지방합성 유전자 발현)

  • 이승환;박응우;조용민;김경훈;오영균;이지혜;이창수;오성종;윤두학
    • Journal of Animal Science and Technology
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    • v.48 no.3
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    • pp.345-352
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    • 2006
  • Korean native cattle (Hanwoo) have a good capacity to produce heavily marbled meat of high value. The intramuscular fat in Hanwoo is known to be deposit from 12 months of age by degree of slightly visible and significantly developed in 28 months of age. Lipogenesis gene expression profiling in longissimus dorsi at early and late fattening stage will be helpful to understand the mechanism of intramuscular fat deposition in skeletal muscle. Therefore, we analysed the gene expression patterns of six genes related lipid metabolism (FABP4, GLUT4, LPL, ACC, ACL and SCD) between early and late fattening stage. The mRNA expression of FABP4 at late fattening stage (27 months old) was higher about 3.0 fold than at early fattening stage (12 months old) in each three individuals of Hanwoo. However, GLUT4 mRNA expression was not different at late fattening stage compared with at early fattening stage. On the other hand, The expression patterns of LPL, ACC, ACL and SCD genes related lipid metabolism were significantly over-expressed about 3.5 fold, 2.7 fold, 3.7 fold and 7.5 fold at late fattening stage, respectively. Thus, these results suggested that lipogenesis in skeletal muscle at late fattening stage is due to increasing uptake of fatty acid by FABP4 and lipogenesis gene expression such as LPL, ACC, ACL and SCD.

Differential gene expression profiles of periodontal soft tissue from rat teeth after immediate and delayed replantation: a pilot study

  • Chae, Yong Kwon;Shin, Seo Young;Kang, Sang Wook;Choi, Sung Chul;Nam, Ok Hyung
    • Journal of Periodontal and Implant Science
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    • v.52 no.2
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    • pp.127-140
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    • 2022
  • Purpose: In dental avulsion, delayed replantation usually has an uncertain prognosis. After tooth replantation, complex inflammatory responses promote a return to periodontal tissue homeostasis. Various types of cytokines are produced in the inflammatory microenvironment, and these cytokines determine the periodontal tissue response. This study aimed to identify the gene expression profiles of replanted teeth and evaluate the functional differences between immediate and delayed replantation. Methods: Maxillary molars from Sprague-Dawley rats were extracted, exposed to a dry environment, and then replanted. The animals were divided into 2 groups according to the extra-oral time: immediate replantation (dry for 5 minutes) and delayed replantation (dry for 60 minutes). Either 3 or 7 days after replantation, the animals were sacrificed. Periodontal soft tissues were harvested for mRNA sequencing. Hallmark gene set enrichment analysis was performed to predict the function of gene-gene interactions. The normalized enrichment score (NES) was calculated to determine functional differences. Results: The hallmark gene sets enriched in delayed replantation at 3 days were oxidative phosphorylation (NES=2.82, Q<0.001) and tumor necrosis factor-alpha (TNF-α) signaling via the nuclear factor kappa light chain enhancer of activated B cells (NF-κB) pathway (NES=1.52, Q=0.034). At 7 days after delayed replantation, TNF-α signaling via the NF-κB pathway (NES=-1.82, Q=0.002), angiogenesis (NES=-1.66, Q=0.01), and the transforming growth factor-beta signaling pathway (NES=-1.46, Q=0.051) were negatively highlighted. Conclusions: Differentially expressed gene profiles were significantly different between immediate and delayed replantation. TNF-α signaling via the NF-κB pathway was marked during the healing process. However, the enrichment score of this pathway changed in a time-dependent manner between immediate and delayed replantation.

Expression of dehydration responsive element-binding protein-3 (DREB3) under different abiotic stresses in tomato

  • Islam, Mohammad Saiful;Wang, Myeong-Hyeon
    • BMB Reports
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    • v.42 no.9
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    • pp.611-616
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    • 2009
  • We investigated the expression pattern of dehydration responsive element-binding protein-3 in tomato under different abiotic stresses. Full length LeDREB3 cDNA was isolated from tomato plant, followed by phylogenetic analysis based on deduced amino acid sequences that revealed significant sequence similarity to DREB proteins belonging to diverse families of plant species. Southern blot analysis showed duplicate copies of LeDREB3 in the tomato genome while organ-specific expression profiling indicated constitutive expression of LeDREB3 in all tested organs, which was particularly strong in flower. LeDREB3 expression was significantly induced by Nacl, drought, low temperature and $H_2O_2$. Moreover, LeDREB3 was slightly regulated by treatment with ABA and MV. These observations suggest that the LeDREB3 gene may be involved in the response of the tomato plant to stress.

Toxin Gene Profiling of Bacillus cereus Food Isolates by PCR

  • Seong, Seon-Je;Lim, Ji-Su;Lee, Kwang-Geun;Lee, Seung-Ju;Hong, Kwang-Won
    • Applied Biological Chemistry
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    • v.51 no.4
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    • pp.263-268
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    • 2008
  • Seventy-one Bacillus cereus strains (12 references and 59 food isolates) were analyzed for the occurrence of five different enterotoxin genes (nheABC, hblCDA, entFM, cytK, and bceT) and one emetic toxin cereulide synthetase gene (ces) by PCR (polymerase chain reaction). PCR analysis revealed eight toxigenic patterns in all B. cereus strains tested; they all carried both entFM and nheABC. The presence of hblCDA, cytK, and bceT varied according to the enterotoxin-producing strains, among which hblCDA was the least frequently detected in the food-isolated strains. Only five B. cereus strains harbored ces, associated with the emetic type of food poisoning; however, these strains were devoid of hblCDA, cytK, and bceT.