• Title/Summary/Keyword: Reporter

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Green Fluorescent Protein-reporter Mammalian One-hybrid System for Identifying Novel Transcriptional Modulators for Human $p14^{ARF}$ Tumor Suppressor Gene

  • Lee, Hye Jin;Yang, Dong Hwa;Yim, Tae Hee;Rhee, Byung Kirl;Kim, Jung-Wook;Lee, Jungwoon;Gim, Jin Bae;Kim, JungHo
    • Animal cells and systems
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    • v.6 no.4
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    • pp.317-322
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    • 2002
  • To improve conventional yeast one-hybrid screening, we have developed an efficient mammalian one-hybrid system that allows rapid isolation of com-plementary DNAs which are able to induce human p14$^{ARF}$. tumor suppressor gene. A 1.5 kb promoter region of p14$^{ARF}$ was fused to EGFP to generate ARF promoter-EGFP reporter vector. This reporter plasmid was stably trans-fected into NIH3T3 cells for generation of reporter cell line. When the reporter cell line was infected with E2F-1 together with excess amounts of empty vector, the cells that received the positive modulator were readily identifiable by green fluorescence using FACS. The GFP-positive cells were cloned directly from the cultured cells and expanded in bulk culture. The genomic DNAs from GFP-positive cells were prepared and the CDNA insert in integrated retroviral genome was recovered by PCR using primers annealing to the retroviral vector sequences flanking the insert-cloning site. This system should be useful for efficient screening of expression CDNA libraries in mammalian cells to identify novel upstream regulators for spe-cific genes by one-hybrid interaction.ion.

Molecular imaging of atherosclerosis using reporter gene system

  • Yoo, Ran Ji;Lee, Kyochul;Kang, Joo Hyun;Lee, Yong Jin
    • Journal of Radiopharmaceuticals and Molecular Probes
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    • v.4 no.1
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    • pp.26-31
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    • 2018
  • Macrophages play a key role in atherosclerotic plaque formation, but their participation has been discerned largely via ex vivo analyses of atherosclerotic lesions. Therefore, we aimed to identify atherosclerosis on noninvasive in vivo imaging using reporter gene system. This study demonstrated that recruitment of macrophages could be detected in atherosclerotic plaques of Apolipoprotein E knockout (ApoE-/-) mice with a sodium iodide symporter (NIS) gene imaging system using $^{99m}Tc-SPECT$. This novel approach to tracking macrophages to atherosclerotic plaques in vivo could have applications in studies of arteriosclerotic vascular disease.

Grp78 is a Novel Downstream Target Gene of Hoxc8 Homeoprotein

  • Kang, Jin-Joo;Bok, Jin-Woong;Kim, Myoung-Hee
    • Biomedical Science Letters
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    • v.17 no.1
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    • pp.1-5
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    • 2011
  • Previously, we have identified 14 putative downstream target genes of Hoxc8 homeoprotein in F9 murine embryonic teratocarcinoma cells through proteomics analysis. Among those, we tested a possibility of a DNA-k type molecular chaperone, Grp78, as a direct downstream target of Hoxc8, by cloning a 2.4 kb upstream region of murine Grp78 into a reporter plasmid and by testing if Hoxc8 can regulate its expression. We observed that Hoxc8 proteins could transactivate the reporter gene, which was affected by small interference RNAs (siRNAs) against to Hoxc8, suggesting that Grp78 is a novel downstream target of Hoxc8 in vivo.

Statistical Analysis Between Size and Balance of Text Corpus by Evaluation of the effect of Interview Sentence in Language Modeling (언어모델 인터뷰 영향 평가를 통한 텍스트 균형 및 사이즈간의 통계 분석)

  • Jung Eui-Jung;Lee Youngjik
    • Proceedings of the Acoustical Society of Korea Conference
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    • spring
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    • pp.87-90
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    • 2002
  • This paper analyzes statistically the relationship between size and balance of text corpus by evaluation of the effect of interview sentences in language model for Korean broadcast news transcription system. Our Korean broadcast news transcription system's ultimate purpose is to recognize not interview speech, but the anchor's and reporter's speech in broadcast news show. But the gathered text corpus for constructing language model consists of interview sentences a portion of the whole, $15\%$ approximately. The characteristic of interview sentence is different from the anchor's and the reporter's in one thing or another. Therefore it disturbs the anchor and reporter oriented language modeling. In this paper, we evaluate the effect of interview sentences in language model for Korean broadcast news transcription system and analyze statistically the relationship between size and balance of text corpus by making an experiment as the same procedure according to varying the size of corpus.

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Directed Evolution of a β-Glucosidase for Improved Functions as a Reporter in Protein Expression

  • Lim, Ho-Dong;Han, So-Young;Park, Gi-Hye;Cheong, Dae-Eun;Kim, Geun-Joong
    • Microbiology and Biotechnology Letters
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    • v.50 no.2
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    • pp.240-244
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    • 2022
  • Precisely reliable and quantitative reporters can provide phenotypes that are consistent with research goals in protein expression. Here, we developed an improved reporter mATglu III 5 by directed evolution using a versatile β-glucosidase ATglu derived from Agrobacterium tumefaciens. When expressed in hosts, a vector containing this mutant distinctly showed a colored or fluorescent phenotype, according to the supplemented substrate, without any inducer. Analysis of mATglu III 5 showed it to be fully functional in fusion state with oligomeric proteins, especially under non-induction conditions, thereby offering an alternative to conventional reporters.

A Study on the Image Expression Technique Analysis of Television News Programs - Mainly on the Image Expression of Camera Reporter and VJ - (텔레비전 뉴스 프로그램의 영상표현기법 분석 - 카메라기자와 VJ의 영상표현을 중심으로 -)

  • Park Dug-Chun
    • The Journal of the Korea Contents Association
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    • v.6 no.8
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    • pp.184-191
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    • 2006
  • The purpose of this case study is to understand the characteristics and implication of videojournalist's image expression techniques in the television news programs, comparing with television camera reporter's image expression techniques. Texts chosen in this thesis are 120 programs of KBS2TV 'News Time' which had been on air from July to December in 2005. Shot size, camerawork, shot duration and camera angle of the images were analysed, which had been extracted through the processes of thematic classification. The results of the analysis of thematic classification shows us that videojournalist covered mainly social, economic and cultural news items, almost completely alienated in the fields of hard news as political and health-scientific news items which were very sensitive and influential on the life of the entire people of the nation. The results of the analysis on the characteristics of image expression reveals that videojournalist used one-shot more often than the camera reporter, showing us videojournalist's preference of close news gathering, and videojournalist used following and zooming much more often than camera reporter, showing us videojoumalist's preference of mobile and dynamic cameraworks, and videojournalist used low angles more often than camera reporter, also showing us videojoumalist's preference of candid camera style news gathering.

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A Molecular Study of Sopungsungi-won(Shufengshunqiyuan) about Regulation of PPARs in Mouse NMu2Li Liver Cells and C2C12 Skeletal Muscle Myogenic Progenital Cells (소풍순기원(疏風順氣元)이 mouse의 NMu2Li 간세포와 C2C12 골격근세포에서 PPARs 조절의 분자기전에 미치는 영향)

  • Oh, Young-Jin;Shin, Soon-Shik;Yoon, Mi-Chung;Kim, Bo-Kyung
    • Journal of Oriental Neuropsychiatry
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    • v.20 no.1
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    • pp.147-164
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    • 2009
  • Objectives : We investigated the effects of Sopungsungi-won(Shu!engshunqiyuan) (SSEx1, SSEx2) to treat the metabolic syndrome by the molecular mechanism of regulation of PPAR and modulation of mitochondrial MCAD, VLCAD mRNA expression. Methods : Mouse NMu2Li liver cells and C2C12 skeletal muscle myogenic progenital cells were transiently transfected with expression plasmids for PPAR(PPAR${\alpha}$, PPAR${\delta}$), a luciferase reporter gene construct containing 3 copies of the PPRE from the rat acyl-CoA oxidase gene and ${\beta}$-galactosidase gene. Cells were treated with several concentrated kinds of SSEx1, SSEx2 at the initial time of culture and analyzed PPAR${\alpha}$, PPAR${\delta}$ reporter gene activity using spectrophotometer (405 nm). Total RNA was extracted from SSEx1, SSEx2 and measured mRNA levels of mitochondrial MCAD, VLCAD. Representative RT-PCR bands are shown. Results : 1. SSEx1 increased the expression of PPAR${\alpha}$ reporter gene activities at 0.1 ${\mu}$g/ml (p${\mu}$g/ml (p<0.05), SSEx2 at 0.1 ${\mu}$g/ml (p${\mu}$g/ml (p<0.05) significantly in NMu2Li liver cell lines. 2. SSEx1 increased the expression of PPAR${\alpha}$ reporter gene activities at 1 ${\mu}$g/ml (p${\mu}$g/ml (p${\alpha}$ reporter gene activities in C2C12 skeletal muscle cells. 4. SSEx1 increased the modulation of mitochondrial MCAD mRNA expression (p<0.05) significantly in NMu2Li liver cell lines. 5. SSEx1, SSEx2 both increased the modulation of mitochondrial MCAD mRNA expression (p<0.05) significantly in C2C12 skeletal muscle cells. Conclusions : These results show the SSEx1, SSEx2 can be used as therapeutic agent for metabolic syndrome and it's molecular mechanisms of PPAR more contribute to the activation of PPAR${\alpha}$ then PPAR${\delta}$ reporter gene activities and it's total RNA more contribute to the modulation of mitochondrial MCAD then VLCAD mRNA expression.

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Mechanism of Phenoxy Compounds as Androgenic Endocrine Disruptors

  • Kim, Hyun-Jung;Park, Yong-In;Dong, Mi-Sook
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2003.10b
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    • pp.165-165
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    • 2003
  • Phenxoy compounds, 2,4-dichlorophenol acetoxyacid (2,4-D) and 2,4-dichlorophenol (DCP), are widely used as a hormonal herbicide and intermediate for pesticide manufacturing, respectively. We have previously reported the potential of these compounds as androgenic endocrine disruptors using in vivo Hershberger assay and in vitro reporter gene assay.(omitted)

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