• Title/Summary/Keyword: Regulatory elements

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Identification of cis-Regulatory Region Controlling Semaphorin-1a Expression in the Drosophila Embryonic Nervous System

  • Hong, Young Gi;Kang, Bongsu;Lee, Seongsoo;Lee, Youngseok;Ju, Bong-Gun;Jeong, Sangyun
    • Molecules and Cells
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    • v.43 no.3
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    • pp.228-235
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    • 2020
  • The Drosophila transmembrane semaphorin Sema-1a mediates forward and reverse signaling that plays an essential role in motor and central nervous system (CNS) axon pathfinding during embryonic neural development. Previous immunohistochemical analysis revealed that Sema-1a is expressed on most commissural and longitudinal axons in the CNS and five motor nerve branches in the peripheral nervous system (PNS). However, Sema-1a-mediated axon guidance function contributes significantly to both intersegmental nerve b (ISNb) and segmental nerve a (SNa), and slightly to ISNd and SNc, but not to ISN motor axon pathfinding. Here, we uncover three cis-regulatory elements (CREs), R34A03, R32H10, and R33F06, that robustly drove reporter expression in a large subset of neurons in the CNS. In the transgenic lines R34A03 and R32H10 reporter expression was consistently observed on both ISNb and SNa nerve branches, whereas in the line R33F06 reporter expression was irregularly detected on ISNb or SNa nerve branches in small subsets of abdominal hemisegments. Through complementation test with a Sema-1a loss-of-function allele, we found that neuronal expression of Sema-1a driven by each of R34A03 and R32H10 restores robustly the CNS and PNS motor axon guidance defects observed in Sema-1a homozygous mutants. However, when wild-type Sema-1a is expressed by R33F06 in Sema-1a mutants, the Sema-1a PNS axon guidance phenotypes are partially rescued while the Sema-1a CNS axon guidance defects are completely rescued. These results suggest that in a redundant manner, the CREs, R34A03, R32H10, and R33F06 govern the Sema-1a expression required for the axon guidance function of Sema-1a during embryonic neural development.

Promoter Analysis of the Cell Surface-abundant and Hypoviral-regulated Cryparin Gene from Cryphonectria parasitica

  • Kim, Myoung-Ju;Kwon, Bo-Ra;Park, Seung-Moon;Chung, Hea-Jong;Yang, Moon-Sik;Churchill, Alice C.L.;Van Alfen, Neal K.;Kim, Dae-Hyuk
    • Molecules and Cells
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    • v.26 no.5
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    • pp.496-502
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    • 2008
  • Cryparin, encoded as a single copy gene (Crp) of the chestnut blight fungus Cryphonectria parasitica, is the most abundant protein produced by this fungus. However, its accumulation is decreased remarkably in C. parastica strains containing the double-stranded (ds) RNA virus Cryphonectria hypovirus 1. To characterize the transcriptional regulatory element(s) for strong expression and viral regulation, promoter analysis was conducted. Serial deletion of the Crp promoter region resulted in a step-wise decrease in promoter activity, indicating a localized distribution of genetic elements in the cryparin promoter. Promoter analysis indicated two positive and a repressive cis-acting elements. Among them, the promoter region between nt -1,282 and -907 appeared to be necessary for hypoviral-mediated down-regulation. An electrophoretic mobility shift assay (EMSA) on the corresponding promoter region (-1,282/-907) indicated two regions at (-1,257/-1,158) and (-1,107/-1,008) with the characteristic AGGAGGA-N42-GAGAGGA and its inverted repeat TCCTCTC-N54-TCCTCCT, respectively, appeared to be specific binding sites for cellular factors.

Identification and Characterization of a Putative Baculoviral Transcriptional Factor IE-1 from Choristoneura fumiferana Granulovirus

  • Rashidan, Kianoush Khajeh;Nassoury, Nasha;Merzouki, Abderrazzak;Guertin, Claude
    • BMB Reports
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    • v.35 no.6
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    • pp.553-561
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    • 2002
  • A gene that encodes a protein homologue to baculoviral IE-1 was identified and sequenced in the genome of the Choristoneura fumiferana granulovirus (ChfuGV). The gene has an 1278 nucleotide (nt) open-reading frame (ORF) that encodes 426 amino acids with an estimated molecular weight of 50.33 kDa. At the nucleotide level, several cis-acting regulatory elements were detected within the promoter region of the ie-1 gene of ChfuGV along with other studied granuloviruses (GVs). Two putative CCAAT elements were detected within the noncoding leader region of this gene; one was located on the opposite strand at -92 and the other at -420 nt from the putative start triplet. Two baculoviral late promoter motifs (TAAG) were also detected within the promoter region of the ie-1 gene of ChfuGV. A single polyadenylation signal, AATAAA, was located 18nt downstream of the putative translational stop codon of ie-1 from ChfuGV. At the protein level, the amino acid sequence data that was derived from the nucleotide sequence in ChfuGV IE-1 was compared to those of the Cydia pomonella granulovirus (CpGV), Xestia c-nigrum granulovirus (XcGV) and Plutella xylostella granulovirus (PxGV). The C-terminal regions of the granuloviral IE-1 sequences appeared to be more conserved when compared to the N-terminal regions. A domain, similar to the basic helix-loop-helix like (bHLH-like) domain in NPVs, was detected at the C-terminal region of IE-1 from ChfuGV (residues 387 to 414). A phylogenetic tree for baculoviral IE-1 was constructed using a maximum parsimony analysis. A phylogenetic estimation demonstrates that ChfuGV IE-1 is most closely related to that of CpGV.

A Methodology for Establishment of Safety Culture in Industry through Global Company Practices

  • Kim, Beom Soo;Lee, Yujeong;Chang, Seong Rok
    • Journal of the Ergonomics Society of Korea
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    • v.35 no.2
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    • pp.85-95
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    • 2016
  • Objective:The purpose of this study is to suggest the methodology for establishment of safety culture in industry. Background: There have been studies indicating positive relation between safety culture and safety performance. But the areas of those efforts have been mainly focused on elements or factors consisting of safety culture. So it is expected that studies on methodologies for developing process of the culture could contribute for the industry to understand and apply it accordingly in order to improve their practices. Method: In this study, the scope was considered just for industry, even though there are many fields of culture. As the first step, the limitations of regulatory control and efforts by industry were reviewed, and the trends of researches were presented. In the second step, the global company practices were provided with detailed activities. Results: This paper showed the limitations of present state in government strategies and industry for safety culture improvement. Also the restricted areas in researches were shown. As one of the solutions to resolve those things, such global company practices as perception to safety journey, the steps of culture development, total safety philosophy, structure and elements of safety process, standards and procedures, training, and up to keys to success were suggested. Conclusion: Through this study, the development of the structure of safety process which consists of many purposeful activities has been derived to be very important. Because it will be the seeds of safety culture expressing their thinking process and behaviors. Also it is required to approach success step by step according to the achievement of each step. Application: This study can be used to develop the methodology and safety process in industry with different viewpoints for safety culture establishment.

Pressure-Overload Cardiac Hypertrophy Is Associated with Distinct Alternative Splicing Due to Altered Expression of Splicing Factors

  • Kim, Taeyong;Kim, Jin Ock;Oh, Jae Gyun;Hong, Seong-Eui;Kim, Do Han
    • Molecules and Cells
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    • v.37 no.1
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    • pp.81-87
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    • 2014
  • Chronic pressure-overload cardiac hypertrophy is associated with an increased risk of morbidity/mortality, largely due to maladaptive remodeling and dilatation that progresses to dilated cardiomyopathy. Alternative splicing is an important biological mechanism that generates proteomic complexity and diversity. The recent development of next-generation RNA sequencing has improved our understanding of the qualitative signatures associated with alternative splicing in various biological conditions. However, the role of alternative splicing in cardiac hypertrophy is yet unknown. The present study employed RNA-Seq and a bioinformatic approach to detect the RNA splicing regulatory elements involved in alternative splicing during pressure-overload cardiac hypertrophy. We found GC-rich exonic motifs that regulate intron retention in 5' UTRs and AT-rich exonic motifs that are involved in exclusion of the AT-rich elements that cause mRNA instability in 3' UTRs. We also identified motifs in the intronic regions involved in exon exclusion and inclusion, which predicted splicing factors that bind to these motifs. We found, through Western blotting, that the expression levels of three splicing factors, ESRP1, PTB and SF2/ASF, were significantly altered during cardiac hypertrophy. Collectively, the present results suggest that chronic pressure-overload hypertrophy is closely associated with distinct alternative splicing due to altered expression of splicing factors.

Delphi Study on Game Content Valuation Model Element Extraction and Analysis (게임 컨텐츠 가치 평가 모델 분석 및 요소 추출에 관한 연구)

  • Lee, Sung Tae;Cho, Ok Hue
    • Journal of the Korean Society for Computer Game
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    • v.31 no.4
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    • pp.145-155
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    • 2018
  • This study aims to extract additional elements to the existing game value evaluation using Delphi technique to 23 game experts. As a result of reviewing the characteristics of the development model using the evaluation model of the existing game contents, it was found that the evaluation factor was selected when selecting the content. The Delphi analysis was conducted two times, the first was the expert FGI and the second was the Delphi questionnaire. More detailed elements were presented in the related items such as results, technology transfer and dissemination, and details for promoting commercialization. Through focus group interviews, we extracted additional factors and predicted market size, growth potential, regulatory and market appeal and risk through market analysis of content success in the game industry. This will allow the industry to understand market size, growth potential, regulation, market appeal and risk through a market analysis of its success targets. This study suggests that the proposed evaluation factors can be applied to the entire game and cultural contents industry and will also help to construct market factors such as market value evaluation of contents, goal and goal setting, selection of entry method, and strategic factors.

Technical Standards and Safety Review of the Low and Intermediate Level Radioactive Waste Disposal Facility (중.저준위 방사성폐기물 처분시설에 대한 기술기준 및 안전심사)

  • Cheong, Jae-Hak;Lee, Kwan-Hee;Lee, Yun-Keun;Jeong, Chan-Woo;Rho, Byung-Hwan
    • Journal of Nuclear Fuel Cycle and Waste Technology(JNFCWT)
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    • v.6 no.4
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    • pp.357-368
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    • 2008
  • On July 31, 2008, the Government issued the construction and operation permit for the first low and intermediate level radioactive waste disposal facility in the Republic of Korea. In this paper, the fundamental regulatory framework, regulatory requirements and technical standards of the disposal facility are introduced, and the phased review process adopted for evaluation of the safety of the facility is briefly described. The Atomic Energy Act sets forth a stepwise regulatory framework for the whole life-cycle of the disposal facility such as siting, design, construction, operation, closure and institutional control. More detailed regulatory requirements and technical standards are stipulated in the subsequent regulations of the Atomic Energy Act and a series of Notices issued by the Ministry of Eduction, Science and Technology. The Korea Institute of Nuclear Safety, as entrusted by the Ministry under the Atomic Energy Act, conducted safety review on the disposal facility, and evaluated the compliance with relevant criteria in all technical elements(i.e. siting and structural safety, radiological environmental impact, operational safety, systems and components, quality assurance, and total systematic performance assessment, etc.). The overall safety review process can be phased into inception phase, initial review phase, main review phase and completion phase. The review results were reported to and deliberated by the five Sub-committees of the Special Committee on Nuclear Safety, and then reported to the Ministry. The Ministry issued the construction and operation permit of the disposal facility through the deliberation of the review results by the Nuclear Safety Commission. Hereafter, the safety of the repository will be reassured by a series of subsequent regulatory inspections and reviews under the Atomic Energy Act. In addition, the licensee's continuous implementation of the "Safety Promotion Plan" may also enhance the long-term safety of the repository and contribute to build-up the confidence of the safety case.

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Epigenetic modification of long interspersed elements-1 in cumulus cells of mature and immature oocytes from patients with polycystic ovary syndrome

  • Pruksananonda, Kamthorn;Wasinarom, Artisa;Sereepapong, Wisan;Sirayapiwat, Porntip;Rattanatanyong, Prakasit;Mutirangura, Apiwat
    • Clinical and Experimental Reproductive Medicine
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    • v.43 no.2
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    • pp.82-89
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    • 2016
  • Objective: The long interspersed elements (LINE-1, L1s) are a group of genetic elements found in large numbers in the human genome that can translate into phenotype by controlling genes. Growing evidence supports the role of epigenetic in polycystic ovary syndrome (PCOS). The purpose of this study is to evaluate the DNA methylation levels in LINE-1 in a tissue-specific manner using cumulus cells from patients with PCOS compared with normal controls. Methods: The study included 19 patients with PCOS and 22 control patients who were undergoing controlled ovarian hyperstimulation. After oocyte retrieval, cumulus cells were extracted. LINE-1 DNA methylation levels were analysed by bisulfite treatment, polymerase chain reaction, and restriction enzyme digestion. The Connection Up- and Down-Regulation Expression Analysis of Microarrays software package was used to compare the gene regulatory functions of intragenic LINE-1. Results: The results showed higher LINE-1 DNA methylation levels in the cumulus cells of mature oocytes in PCOS patients, 79.14 (${\pm}2.66$) vs. 75.40 (${\pm}4.92$); p=0.004, but no difference in the methylation of cumulus cells in immature oocytes between PCOS and control patients, 70.33 (${\pm}4.79$) vs. 67.79 (${\pm}5.17$); p=0.155. However, LINE-1 DNA methylation levels were found to be higher in the cumulus cells of mature oocytes than in those of immature oocytes in both PCOS and control patients. Conclusion: These findings suggest that the epigenetic modification of LINE-1 DNA may play a role in regulating multiple gene expression that affects the pathophysiology and development of mature oocytes in PCOS.

Molecular Cloning and Characterization of Mn-Superoxide Dismutase Gene from Candida sp.

  • Hong, Yun-Mi;Nam, Yong-Suk;Choi, Soon-Yong
    • Journal of Microbiology
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    • v.35 no.4
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    • pp.309-314
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    • 1997
  • The manganese-containing superoxide dismutase (MnSOD) is a major component of the cellular defence mechanisms against the toxic effects of the superoxide radical. Within the framework of studies on oxidative stress=responsible enzymes in the Candida sp., the gene encoding the MnSOD was isolated and examined in this study. A specific primer was designed based on conserved regions of MnSOD sequences from other organisms, and was used to isolate the gene by PCR on reverse-transcribed Candida poly($A^{+}$) RNA. The PCR product was used to screen a Candida genomic lambda library and the nucleotide wequence of positive clone was determined. The deduced primary sequence encodes a 25kDa protein which has the conserved residues for enzyme activity and metal binding. The 28 N-terminal amino acids encoded by the Candida cDNA comprise a putatice mitochondrial transit peptide. Potential regulatory elements were identified in the 5' flanking sequences. Northern blot analysis showed that the transcription of the MnSOD gene is induced 5-to 10-fold in response to mercury, cadmium ions and hydrogen peroxide.

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The Role of High-throughput Transcriptome Analysis in Metabolic Engineering

  • Jewett, Michael C.;Oliveira, Ana Paula;Patil, Kiran Raosaheb;Nielsen, Jens
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.5
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    • pp.385-399
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    • 2005
  • The phenotypic response of a cell results from a well orchestrated web of complex interactions which propagate from the genetic architecture through the metabolic flux network. To rationally design cell factories which carry out specific functional objectives by controlling this hierarchical system is a challenge. Transcriptome analysis, the most mature high-throughput measurement technology, has been readily applied In strain improvement programs in an attempt to Identify genes involved in expressing a given phenotype. Unfortunately, while differentially expressed genes may provide targets for metabolic engineering, phenotypic responses are often not directly linked to transcriptional patterns, This limits the application of genome-wide transcriptional analysis for the design of cell factories. However, improved tools for integrating transcriptional data with other high-throughput measurements and known biological interactions are emerging. These tools hold significant promise for providing the framework to comprehensively dissect the regulatory mechanisms that identify the cellular control mechanisms and lead to more effective strategies to rewire the cellular control elements for metabolic engineering.