• Title/Summary/Keyword: Reporter

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Versatile Luciferase Reporter Plasmids for Transcription Studies in Diverse Eukaryotic Cells (다양한 진핵생물 세포에서 전사 연구에 사용될 수 있는 Luciferase Reporter Plasmid의 개발)

  • 조영석;한동욱;백금희;박승필;윤상순;임운기;김정락;김한도;강호성
    • The Korean Journal of Zoology
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    • v.39 no.4
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    • pp.378-386
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    • 1996
  • We have developed a couple of new luciferase reporter plasmida with very low background reporter actlvltlea. One can be used to measure the promoter strength, after insertion of some promoter fragment into the reporter plasmid, and the other, with very low basal promoter actlvltlea, allis In studying eukaryotic transcriptional regulators. The latter reporter plasmid contains such cli elements as a 17 nucleotide long inftlator, Spl.blndIng sftes, GAL4 binding sltea, and bInding sitea for a certain Drosophila homeodomain proteins. In an attempt to construct an improved reporter plaimid by fadlltating transcriptional termination and minimizing any interference by cryptic promoters which may be preaent in the reporter pleamld DNA, we have inserted transsrlptional termination-related signals, a three tandem repeat of SV4O polyadenylatlon signal (AAA) and the putative transcrtptional termination signal (UMS) of the mouse c-mos gene, Into just upstream of the initIator, and the promoter actlvitiea were measured by a transIent expression assay employing the Drosophila Schneider line 2 cells. As expected, the basal promoter activitIes decreased maximally when both transcription termination related elements were inserted. Moreover, the reporter plasmld with the two elements allowed more sensitive measurement of transcriptional activation than the reporter piasmid without them. Theae reporter plasmids can be used for studying transcriptional regulators of higher organisms Including mammals as well as Droiophlla melanogaiter.

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Establishment of Reporter Cell Lines that Monitor Activities of Hypoxia Inducible Factor-1, P53 and Nur77 for Assessment of Carcinogenicity (저산소유도인자 HIF-1, 암 억제인자 P53과 고아 핵수용체 Nur77의 발현을 지표로 하는 발암독성예측 세포주의 개발)

  • Hong, Il;Seo, Hee-Won;Lee, Min-Ho;Kim, Ji-Won;Chung, Jin-Ho;Lee, Byung-Hoon;Lee, Mi-Ock
    • Toxicological Research
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    • v.23 no.3
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    • pp.231-238
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    • 2007
  • Evaluation of potentials of chemicals to alter expression of genes that are involved in carcinogenesis may serve useful tools in toxicological research. In this investigation, we developed reporter cell lines that expressed luciferase in response to transactivation of hypoxia inducible factor-1, P53 tumor suppressor and Nur77 of which roles have been well established in cancer development and progression. Whereas these reporter cell lines displayed low constitutive backgrounds, the reporter activities were significantly enhanced in response to $desferriosamine/CoCl_2$, adriamycin or 6-mercaptopurine, which are hypoxia mimicking chemicals, P53 activator or Nur77 inducer, respectively. The activation of the reporter was time- and dose-dependent. Known tumor initiators and promoters, such as phorbol 12-myristate 13-acetate and phorbol 12, 13-dicaprinate induced the reporter activity at as low as 10nM in these stable cell lines. Further, known anti-tumor promoters, such as ascorbic acid and ${\beta}-carotene$ repressed the reporter activities. These results indicate that our stable reporter cell lines could serve as a useful system for rapid assessment of carcinogenicity of toxic chemicals.

The art of reporter proteins in science: past, present and future applications

  • Ghim, Cheol-Min;Lee, Sung-Kuk;Takayama, Shuichi;Mitchell, Robert J.
    • BMB Reports
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    • v.43 no.7
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    • pp.451-460
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    • 2010
  • Starting with the first publication of lacZ gene fusion in 1980, reporter genes have just entered their fourth decade. Initial studies relied on the simple fusion of a promoter or gene with a particular reporter gene of interest. Such constructs were then used to determine the promoter activity under specific conditions or within a given cell or organ. Although this protocol was, and still is, very effective, current research shows a paradigm shift has occurred in the use of reporter systems. With the advent of innovative cloning and synthetic biology techniques and microfluidic/nanodroplet systems, reporter genes and their proteins are now finding themselves used in increasingly intricate and novel applications. For example, researchers have used fluorescent proteins to study biofilm formation and discovered that microchannels develop within the biofilm. Furthermore, there has recently been a "fusion" of art and science; through the construction of genetic circuits and regulatory systems, researchers are using bacteria to "paint" pictures based upon external stimuli. As such, this review will discuss the past and current trends in reporter gene applications as well as some exciting potential applications and models that are being developed based upon these remarkable proteins.

Construction of a Bioluminescent Reporter Using the luc Gene and meta-Cleavage Dioxygenase Promoter for Detection of Catecholic Compounds

  • Park, Sang-Ho;Lee, Dong-Hun;Oh, Kye-Heon;Kim, Chi-Kyung
    • Journal of Microbiology
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    • v.38 no.3
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    • pp.183-186
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    • 2000
  • Several types of bioluminescent reporter strains have been developed for the detection and monitoring of pollutant aromatics contaminating the environment. In this study, a bioluminescent reporter strain, E. coli SHP3, was constructed by fusing the luc gene of firefly luciferase with the promoter of pcbC responsible for the meta-cleavage of aromatic hydrocarbons. the bioluminescence expressed by the luc gene in the reporter was well triggered by the promoter when it was exposed to 2,3-dihydroxybiphenyI (2,3-DHBP) at 0.5 to 1 mM concentrations. The bioluminescent response was more extensive when the reporter strain was exposed to 5 mM catechol and 2 mM 4-chlorocatechol. These different types of bioluminescent responses by E. coli SHP3 appeared to be characterized by the nature of the aromatics to stress. Since E. coli SHP3 responded to 2,3-DHBP quite sensitively, this reporter strain could be applied for detecting some catecholic pollutants.

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Journalistic Differences between Blogs of Professional Reporter and Citizen Reporter: Focused on Watchdog and Interactivity (전문기자와 시민기자 블로그 콘텐츠의 저널리즘적 특성에 관한 비교 연구: 감시견과 상호작용성을 중심으로)

  • Kim, Min-Ha;Shin, Yun-Kyoung
    • Korean journal of communication and information
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    • v.53
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    • pp.73-99
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    • 2011
  • This study compares blogs of professional reporters with those of citizen reporters in terms of watchdog and interactivity functions. Watchdog function was analyzed in light of the extent of soft news and the degree of critical relationship with the government. Interactivity was assessed by analyzing readers' comments on the articles of reporter blogs. for citizen journalism and for professional one were chosen in order to minimize any discrepancies caused by ideological differences. As a result of the content analyses, citizen reporter blogs were found to deliver soft news more frequently than those of , whereas the former had stronger tendency to maintain the critical relationship with the government than the latter. As for the interactivity function, although the number of comments uploaded to citizen reporter blogs was higher than that of professional reporter blogs, the latter was found to meet the standards of communicative interaction more adequately than the former.

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Exploring Subjectivity Concerning Occupational Achievements of KBS Journalists (KBS 기자의 직업적 성취에 관한 주관성 탐색)

  • Chung, Je Hyuk;Yoo, Sang Won
    • The Journal of the Korea Contents Association
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    • v.21 no.1
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    • pp.227-242
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    • 2021
  • It is common to say that Korean journalists' professionalism including the public broadcaster KBS is immature. This study used the Q methodology to explore the subjectivity that KBS reporters and citizens have formed about reporters' occupational achievements. we tried to find out the background and reasons of the subjective dimensions of journalists regarding the immature state of Korean journalists' professionalism. Under a condition for sorting of 'a reporter most likely to be promoted', using 95 statements related to the reporter's occupational achievements, 30 KBS reporters, 10 university students who were eager to be a journalist, and 40 ordinary people of various ages, a total of 80 participants took part in the Q sorting. A total of 3 Q factors were extracted, then analyzed in detail into a total of 6 reporter types divided into (+) and (-) sub types for each factor. As a result, six types of perceptions and attitudes were named as 'a professional reporter','a schemer reporter','an organizational loyalty reporter', 'a passionate reporter','a careerist reporter', and 'a reliable worker'. This study can be meaningful in that it further deepened and expanded the existing researches concerning Korean journalist's professional identity.

Imaging Gene Expression (유전자 발현 영상기법)

  • Lee, Kyung-Han
    • The Korean Journal of Nuclear Medicine
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    • v.34 no.1
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    • pp.1-9
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    • 2000
  • The rapid progress of molecular genetic methods over the past two decades has necessitated the development of methods to detect and quantify genetic activity within living bodies. Reporter genes provide a rapid and convenient tool to monitor gene expression by yielding a readily measurable phenotype upon expression when introduced into a biological system. Conventional reporter systems, however, are limited in their usefulness for in vivo experiments or human gene therapy because of its invasive nature which requires cell damage before assays can be performed. This offers an unique opportunity for nuclear imaging techniques to develope a novel method for imaging both the location and amount of gene expression noninvasively. Current developments to achieve this goal rely on utilizing either reporter enzymes that accumulate radiolabeled substrates or reporter receptors that bind specific radioligands. This overview includes a brief introduction to the background for such research, a summary of published results, and an outlook for future directions.

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A New Reporter Vector System Based on Flow-Cytometry to Detect Promoter Activity

  • Jung, Sun-Do;Choi, Ji-Hye;Hong, Chang-Wan;Lee, Hyun-Ji;Park, Yoon-Kyung;Shin, Jung-Hoon;Park, Jae-Won;Park, Se-Ho
    • IMMUNE NETWORK
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    • v.9 no.6
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    • pp.243-247
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    • 2009
  • In this study, we report the development of a new dual reporter vector system for the analysis of promoter activity. This system employs green fluorescence emitting protein, EGFP, as a reporter, and uses red fluorescence emitting protein, DsRed, as a transfection control in a single vector. The expression of those two proteins can be readily detected via flow cytometry in a single analysis, with no need for any further manipulation after transfection. As this system allows for the simultaneous detection of both the control and reporter proteins in the same cells, only transfected cells which express the control protein, DsRed, can be subjected to promoter activity analysis, via the gating out of all un-transfected cells. This results in a dramatic increase in the promoter activity detection sensitivity. This novel reporter vector system should prove to be a simple and efficient method for the analysis of promoter activity.

Development of a Reporter System for In Vivo Monitoring of γ-Secretase Activity in Drosophila

  • Hong, Young Gi;Roh, Seyun;Paik, Donggi;Jeong, Sangyun
    • Molecules and Cells
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    • v.40 no.1
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    • pp.73-81
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    • 2017
  • The ${\gamma}$-secretase complex represents an evolutionarily conserved family of transmembrane aspartyl proteases that cleave numerous type-I membrane proteins, including the ${\beta}$-amyloid precursor protein (APP) and the receptor Notch. All known rare mutations in APP and the ${\gamma}$-secretase catalytic component, presenilin, which lead to increased amyloid ${\beta}$-peptide production, are responsible for early-onset familial Alzheimer's disease. ${\beta}$-amyloid protein precursor-like (APPL) is the Drosophila ortholog of human APP. Here, we created Notch- and APPL-based Drosophila reporter systems for in vivo monitoring of ${\gamma}$-secretase activity. Ectopic expression of the Notch- and APPL-based chimeric reporters in wings results in vein truncation phenotypes. Reporter-mediated vein truncation phenotypes are enhanced by the Notch gain-of-function allele and suppressed by RNAi-mediated knockdown of presenilin. Furthermore, we find that apoptosis partly contributes to the vein truncation phenotypes of the APPL-based reporter, but not to the vein truncation phenotypes of the Notch-based reporter. Taken together, these results suggest that both in vivo reporter systems provide a powerful genetic tool to identify genes that modulate ${\gamma}$-secretase activity and/or APPL metabolism.

Moving Object Tracing System using Network-connected CCTV and Smartphone (네트워크 CCTV와 스마트 단말기를 연동한 이동체 추적 시스템)

  • Lim, Seungkyun;Lee, Sangwook
    • The Journal of the Korea Contents Association
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    • v.13 no.12
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    • pp.519-529
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    • 2013
  • This paper introduces a moving object tracing system using network-connected CCTV and smartphone. In an emergency situation, a smartphone of the reporter gains its GPS information and sends that to the central server. The central server stores received GPS information as a dangerous area in the database and keeps sending the order of tracing the reporter to a network-connected CCTV which can film the reporter. At the same time, the central server sends pictures of the reporter to his or her family and related organization in order to handle the emergency situation as soon as possible. In addition, when a reporter want to know the risk around destination, the central server informs dangerous areas to the reporter by using smart phone application and database of a danger spot.