• Title/Summary/Keyword: cis-element

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Characterization of a Positive Regulatory cis-Element and Transacting Factors for the Hepatitis B Viral Pregenomic Promoter

  • Choi, Cheol-Yong;Park, Geon-Tae;Rho, Hyune-Mo
    • BMB Reports
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    • v.29 no.2
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    • pp.156-162
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    • 1996
  • Transcription of hepatitis B viral pregenomic promoter is known to be regulated mainly by the combined interaction of enhancers I, II and the intervening regulatory sequences between the two enhancers. A positive regulatory element was identified by serial deletion and measuring the linked chloramphenicol acetyltransferase (CAT) activities, which overlapped with the 5' region of the X open reading frame. When the positive regulatory element was inserted upstream of the SV40 early promoter, it elevated SV40 promoter activity in HepG2 cells. Two cellular proteins of 110 (p110) and 33 (p33) kDa interacted with the positive element and both of them were present in the nucleus, but p110 also existed in the cytoplasm in phosphorylated form. Dephosphorylation of p110 by acid phosphatase enhanced the DNA-binding activity of p110. The p33 could bind to single-strand DNA specifically as well as to double-strand DNA.

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Identification of a cis-acting Element Region in the Promoter of Porcine Uroplakin II Gene

  • Kwon, Deug-Nam;Kim, Jin-Hoi
    • Proceedings of the KSAR Conference
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    • 2004.06a
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    • pp.194-194
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    • 2004
  • Tissue-specific expression of the desired gene product in the targrt tissue is central to the concept of bioreactor. One approach is to use a tissue-specific promoter to drive desired gene. To investigate the feasibility of tissue-specific gene expression for bladder using the porcine uroplakin(UPII) promoter and its transcriptional control the efficacy of this promoter as well as well as fragments in regulating gene expression were cell lines using DNA transfection. (omitted)

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Identification of the Regulatory Region Responsible for Vascular Tissue-Specific Expression in the Rice Hd3a Promoter

  • Pasriga, Richa;Cho, Lae-Hyeon;Yoon, Jinmi;An, Gynheung
    • Molecules and Cells
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    • v.41 no.4
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    • pp.342-350
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    • 2018
  • Flowering time is determined by florigens. These genes include, Heading date 3a (Hd3a) and Rice FT 1 (RFT1) in rice, which are specifically expressed in the vascular tissues of leaves at the floral transition stage. To study the cis-regulatory elements present in the promoter region of Hd3a, we generated transgenic plants carrying the 1.75-kb promoter fragment of Hd3a that was fused to the ${\beta}$-glucuronidase (GUS) reporter gene. Plants expressing this construct conferred a vascular cell-specific expression pattern for the reporter gene. However, GUS was expressed in leaves at all developmental stages, including the early seedling stage when Hd3a was not detected. Furthermore, the reporter was expressed in roots at all stages. This suggests that the 1.75-kb region lackings cis-elements that regulate leaf-specific expression at the appropriate developmental stages. Deletion analyses of the promoter region indicated that regulatory elements determining vascular cell-specific expression are present in the 200-bp region between -245 bp and -45 bp from the transcription initiation site. By transforming the Hd3a-GUS construct to rice cultivar 'Taichung 65' which is defective in Ehd1, we observed that Ehd1 is the major regulatory element that controls Hd3a promoter activity.

A Study on the Development of CAD System for VFD Element Tools (형광 표시관 부품의 금형 자동설계 시스템에 관한 연구)

  • 박상봉
    • Proceedings of the Korean Society of Precision Engineering Conference
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    • 1997.04a
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    • pp.724-728
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    • 1997
  • A CAD system of grid element for vacuum fluorescent disply has been developed. In order to reduce design man-houre and human erros, it is used to automate the design process using a knowledge base system. In the case of VFD product design, the most important consideration is the short-life cycle. So the development of CAD system for VFD product is needed. The developed system is based on the knowledge base system which is involved in a lot of expert's technology in the practice field. Using C-language under the HP-UNIX system, CIS customer language of the EXCESS CAD/CAM is used as the overall CAD environment. Results of this system will provide effective aids to the designer in this field

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An Expert System of Grid Die Auto Design for Electron Gun (전자총 전극 금형 자동설계 전문가 시스템)

  • 박상봉
    • Journal of the Korean Society for Precision Engineering
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    • v.15 no.10
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    • pp.27-33
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    • 1998
  • Due to the requirement of high precision and complexity during the die design for electron gun elements, it sometimes causes human-errors. In addition, in the case of beginners, they have more trial and error than skilled engineers. To remove these problems and increase the productivity in the practice field, it needs the development of CAD system for electron gun element progressive die. Through this study, a CAD system of electron gun element progressive die, which is used, by knowledge base system including expert's technology in the practice, has been developed. To develop this system, it has used C-language under HP-UNIX system and CIS customer language of the EXCESS CAD/CAM system. An application of this system will provide effective aids to the designer in this field.

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Structural Characteristics of Two Wheat Histone H2A Genes Encoding Distinct Types of Variants and Functional Differences in Their Promoter Activity

  • Huh, Gyung-Hye;Iwabuchi, Masaki
    • Proceedings of the Botanical Society of Korea Conference
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    • 1996.07a
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    • pp.26-38
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    • 1996
  • To investigate the regulation of plant histone H2A gene expression, we isolated two H2A genes (TH254 and TH274) from wheat, which encode different types of variants. Both genes had an intron in the coding region. In the promoters, some characteristics sequences, such as Oct and Nona motifs, which are conserved among plant histone genes were also found, and they were located in a short region (about 120 bp) upstream from the putative TATA box. Analyses of promoter activity with H2A-GUS fusion genes in the transient system using tobacco protoplasts revealed novel types of positive cis-acting sequences in the TH254 promoter: a direct repeat of a 13-bp sequence (AGTTACATTATTG) and a stretch composed of an AT-rich sequence (ATATAGAAAATTAAAA) and a G-box (CACGTG). A quantitative S1 assay of the mRNA amounts from the TH254/GUS and TH274/GUG chimeric genes in stably transformed and cell cycle-synchronized tobacco cell lines showed that the promoters of both genes contained at least one cis-acting element responsible for S phase-specific expression. Histochemical analysis of transgenic tobacco plants carrying the chimeric genes showed that the promoters of the two H2A genes were both active in developing seedlings and flower organs but regulated in different manner.

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Roles of RUNX1 and PU.1 in CCR3 Transcription

  • Su-Kang Kong;Byung Soo Kim;Sae Mi Hwang;Hyune Hwan Lee;Il Yup Chung
    • IMMUNE NETWORK
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    • v.16 no.3
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    • pp.176-182
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    • 2016
  • CCR3 is a chemokine receptor that mediates the accumulation of allergic inflammatory cells, including eosinophils and Th2 cells, at inflamed sites. The regulatory sequence of the CCR3 gene, contains two Runt-related transcription factor (RUNX) 1 sites and two PU.1 sites, in addition to a functional GATA site for transactivation of the CCR3 gene. In the present study, we examined the effects of the cis-acting elements of RUNX1 and PU.1 on transcription of the gene in EoL-1 eosinophilic cells and Jurkat T cells, both of which expressed functional surface CCR3 and these two transcription factors. Introduction of RUNX1 siRNA or PU.1 siRNA resulted in a modest decrease in CCR3 reporter activity in both cell types, compared with transfection of GATA-1 siRNA. Cotransfection of the two siRNAs led to inhibition in an additive manner. EMSA analysis showed that RUNX1, in particular, bound to its binding motifs. Mutagenesis analysis revealed that all point mutants lacking RUNX1- and PU.1-binding sites exhibited reduced reporter activities. These results suggest that RUNX1 and PU.1 participate in transcriptional regulation of the CCR3 gene.

Cis-acting Replication Element Variation of the Foot-and-mouth Disease Virus is Associated with the Determination of Host Susceptibility (구제역바이러스의 숙주 특이성 결정에 연관되어있는 구제역바이러스 cis-acting replication element 변이 분석 연구)

  • Kang, Hyo Rin;Seong, Mi So;Ku, Bok Kyung;Cheong, JaeHun
    • Journal of Life Science
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    • v.30 no.11
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    • pp.947-955
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    • 2020
  • The foot-and-mouth disease virus (FMDV), a member of the Aphthovirus genus in the Picornaviridae family, affects wild and domesticated ruminants and pigs. During replication of the FMDV RNA (ribonucleic acid) genome, FMDV-encoding RNA polymerase 3D acts in a highly location-specific manner. This suggests that specific RNA structures recognized by 3D polymerase within non-coding regions of the FMDV genome assist with binding during replication. One such region is the cis-acting replication element (CRE), which functions as a template for RNA replication. The FMDV CRE adopts a stem-loop conformation with an extended duplex stem, supporting a novel 15-17 nucleotide loop that derives stability from base-stacking interactions, with the exact RNA nucleotide sequence of the CRE producing different RNA secondary structures. Here, we show that CRE sequences of FMDVs isolated in Korea from 2010 to 2017 exhibit A and O genotypes. Interestingly, variations in the RNA secondary structure of the Korean FMDVs are consistent with the phylogenetic relationships between these viruses and reveal the specificity of FMDV infections for particular host species. Therefore, we conclude that each genetic clade of Korean FMDV is characterized by a unique functional CRE and that the evolutionary success of new genetic lineages may be associated with the invention of a novel CRE motif. Therefore, we propose that the specific RNA structure of a CRE is an additional criterion for FMDV classification dependent on the host species. These findings will help correctly analyze CRE sequences and indicate the specificity of host species for future FMDV epidemics.

An Important Role of Nrf2-ARE Pathway in the Cellular Defense Mechanism

  • Lee, Jong-Min;Johnson, Jeffrey A.
    • BMB Reports
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    • v.37 no.2
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    • pp.139-143
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    • 2004
  • The antioxidant responsive element (ARE) is a cis-acting regulatory element of genes encoding phase II detoxification enzymes and antioxidant proteins, such as NAD(P)H: quinone oxidoreductase 1, glutathione S-transferases, and glutamate-cysteine ligase. Interestingly, it has been reported that Nrf2 (NF-E2-related factor 2) regulates a wide array of ARE-driven genes in various cell types. Nrf2 is a basic leucine zipper transcription factor, which was originally identified as a binding protein of locus control region of ss-globin gene. The DNA binding sequence of Nrf2 and ARE sequence are very similar, and many studies demonstrated that Nrf2 binds to the ARE sites leading to up-regulation of downstream genes. The function of Nrf2 and its downstream target genes suggests that the Nrf2-ARE pathway is important in the cellular antioxidant defense system. In support of this, many studies showed a critical role of Nrf2 in cellular protection and anti-carcinogenicity, implying that the Nrf2-ARE pathway may serve as a therapeutic target for neurodegenerative diseases and cancers, in which oxidative stress is closely implicated.