• Title/Summary/Keyword: Gal2

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Improvement of the Biosensor for Detection of Endocrine Disruptors by Combination of Human Estrogen Receptorα and Co-Activator (Human Estrogen Receptor α와 Co-activator로 구성된 바이오센서를 이용한 내분비계장애물질의 검출)

  • Lee, Haeng-Seog
    • Journal of Korean Society of Water and Wastewater
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    • v.20 no.6
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    • pp.893-904
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    • 2006
  • To improve sensitivity of biosensor as yeast two-hybrid detection system for estrogenic activity of suspected chemicals, we tested effects of several combinations of the bait and fish components in the two-hybrid system on Saccharomyces cerevisiae inducted a chromosome-integrated lacZ reporter gene that was under the control of CYC1 promoter and the upstream Gal4p-binding element $UAS_{GAL}$. The bait components that were fused with the Gal4p DNA binding domain are full-length human estrogen receptor ${\alpha}$ and its ligand-binding domain. The fish components that were fused with the Gal4p transcriptional activation domain were nuclear receptor-binding domains of co-activators SRC1 and TIF2. We found that the combination of the full-length human estrogen receptor ${\alpha}$ with the nuclear receptor-binding domain of co-activator SRC1 was most effective for the estrogen-dependent induction of reporter activity among the two-hybrid systems so far reported. The relative strength of transcriptional activation by representative natural and xenobiotic chemicals was well correlated with their estrogenic potency that had been reported with other assay systems.

Computation of Complete Bouguer Anomalies from Free-air Anomalies in East Sea (동해 지역의 프리에어 이상으로부터 완전부우게 이상의 계산)

  • Yun, Hong-Sik;Lee, Dong-Ha;Kim, Young-Hyun
    • Journal of the Korean Society of Surveying, Geodesy, Photogrammetry and Cartography
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    • v.28 no.3
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    • pp.317-328
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    • 2010
  • This paper describes the results of complete Bouguer anomalies computed from the Free-air anomalies that derived from Sandwell and DNSC08 marine gravity models. Complete bouguer corrections consist of three parts: the bouguer correction (Bullard A), the curvature correction (Bullard B) and the terrain correction (Bullard C). These all corrections have been computed over the East Sea on a $1'{\times}1'$elevation data (topography and bathymetry) derived from ETOPO1 global relief model. In addition, a constant topographic (sea-water) density of $2,670kg/m^3$($1,030kg/m^3$) has been used for all correction terms. The distribution of complete bouguer anomalies computed from DNSC08 are -34.390 ~ 267.925 mGal, and those from Sandwell are -32.446 ~ 266.967 mGal in East Sea. The mean and RMSE value of the difference between DNSC08 and Sandwell is $0.036{\pm}2.373\;mGal$. The highest value of complete bouguer anomaly are found around the region of $42{\sim}43^{\circ}N$ and $137{\sim}139^{\circ}E$ (has the lowest bathymetry) in both models. These values show that the gravity distribution of both models, DNSC08 and Sandwell, are very similar. They indicate that satellite-based marine gravity model can be effectively used to analyze the geophysical, geological and geodetic characteristics in East Sea.

Bitter taste receptors protect against skin aging by inhibiting cellular senescence and enhancing wound healing

  • Chung, Min Gi;Kim, Yerin;Cha, Yeon Kyung;Park, Tai Hyun;Kim, Yuri
    • Nutrition Research and Practice
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    • v.16 no.1
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    • pp.1-13
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    • 2022
  • BACKGROUND/OBJECTIVES: Bitter taste receptors are taste signaling pathway mediators, and are also expressed and function in extra-gustatory organs. Skin aging affects the quality of life and may lead to medical issues. The purpose of this study was to better understand the anti-skin aging effects of bitter taste receptors in D-galactose (D-gal)-induced aged human keratinocytes, HaCaT cells. MATERIALS/METHODS: Expressions of bitter taste receptors in HaCaT cells and mouse skin tissues were examined by polymerase chain reaction assay. Bitter taste receptor was overexpressed in HaCaT cells, and D-gal was treated to induce aging. We examined the effects of bitter taste receptors on aging by using β-galactosidase assay, wound healing assay, and Western blot assay. RESULTS: TAS2R16 and TAS2R10 were expressed in HaCaT cells and were upregulated by D-gal treatment. TAS2R16 exerted protective effects against skin aging by regulating p53 and p21, antioxidant enzymes, the SIRT1/mechanistic target of rapamycin pathway, cell migration, and epithelial-mesenchymal transition markers. TAS2R10 was further examined to confirm a role of TAS2R16 in cellular senescence and wound healing in D-gal-induced aged HaCaT cells. CONCLUSIONS: Our results suggest a novel potential preventive role of these receptors on skin aging by regulating cellular senescence and wound healing in human keratinocyte, HaCaT.

Expression of ${\alpha}$-Galactosidase Gene from Leuconostoc mesenteroides SY1 in Lactobacillus brevis 2.14

  • Lee, Kang-Wook;Park, Ji-Yeong;Park, Jae-Yong;Chun, Ji-Yeon;Kim, Jeong-Hwan
    • Food Science and Biotechnology
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    • v.17 no.5
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    • pp.1115-1118
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    • 2008
  • ${\alpha}$-Galactosidase gene (aga) from Leuconostoc mesenteroides SY1 was expressed in a heterologous host, Lactobacillus brevis 2.14 using an Escherichia coli-Leuconostoc shuttle vector, pSJE. pSJEaga (pSJE carrying aga) was introduced into Lactobacillus brevis 2.14 by electroporation and transformation efficiency was $1.1{\times}10^3$ per ${\mu}g$ DNA. L. brevis transformants (TFs) showed higher ${\alpha}$-galactosidase (${\alpha}$-Gal) activities than cells containing pSJE. Transcription levels of aga in L. brevis 2.14 grown on different carbon sources (1%, w/v) were examined by slot blot analysis. Aga transcript levels and ${\alpha}$-Gal activities were higher in cells grown on melibiose, raffinose, and galactose than cells on glucose, sucrose, and fructose. Western blot result showed that L. brevis 2.14 harboring pSJEaga produced much more ${\alpha}$-Gal when grown on melibiose than on glucose.

Regulation of Nrf2 Transactivation Domain Activity by p160 RAC3/SRC3 and Other Nuclear Co-Regulators

  • Lin, Wen;Shen, Guoxiang;Yuan, Xiaoling;Jain, Mohit R.;Yu, Siwang;Zhang, Aihua;Chen, J. Don;Kong, Ah-Ng Tony
    • BMB Reports
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    • v.39 no.3
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    • pp.304-310
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    • 2006
  • Transcription factor NF-E2-related factor 2 (Nrf2) regulates the induction of Phase II detoxifying enzymes and antioxidant enzymes in response to many cancer chemopreventive compounds. In this study, we investigated the role of receptor associated coactivator (RAC3) or steroid receptor coactivator-3 (SRC3) and other nuclear co-regulators including CBP/p300 (CREB-binding protein), CARM1 (Coactivator-associated arginine methyltransferase), PRMT1 (Protein arginine methyl-transferase 1), and p/CAF (p300/CBP-associated factor) in the transcriptional activation of a chimeric Gal4-Nrf2-Luciferase system containing the transactivation domain (TAD) of Nrf2 in HepG2 cells. The results indicated that RAC3 up-regulated the transactivation activity of Gal4-Nrf2-(1-370) in a dose-dependent manner. The enhancement of transactivation domain activity of Gal4-Nrf2-(1-370) by RAC3 was dampened in the presence of dominant negative mutants of RAC3. Next we studied the effects of other nuclear co-regulators including CBP/p300, CARM1, PRMT1 and p/CAF, and the results showed that they had different level of positive effects on this transactivation domain activity of Gal4-Nrf2-(1-370). But importantly, synergistic effects of these co-regulators in the presence of RAC3/SRC3 on the transactivation activity of Gal4-Nrf2-(1-370) were observed. In summary, our present study showed for the first time that the 160 RAC3/SRC3 is involved in the functional transactivation of TAD of Nrf2 and that the other nuclear co-regulators such as CBP/p300, CARM1, PRMT1 and p/CAF can also transcriptionally activate this TAD of Nrf2 and that they could further enhance the transactivation activity mediated by RAC3/SRC3.

A plasmid vector faciliting gene expression in both yeast and mammalian cells

  • Lee, Tae-Ho
    • Journal of Microbiology
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    • v.35 no.2
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    • pp.149-151
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    • 1997
  • A plasmid vector with combined features of yeast shuttle vector and mammalian expression vector was constructed to facilitate expression of cloned gene in both cell-types. All necessary elements required for plasmid maintenance and selection in E. coli, yeast and mammalian cells were size-economically arranged in this plasmid. The numan cytomegalovirus (CMV) immediate early promoter and yeast GAL1 promoter were sequentially placed in front of the gene to be expressed. The synthetic splicing donor and acceptor sequences were inserted into the immediate upstream and downstream of the GAL1 promotor, allowing the CMV promotor to direct the expression of a given gene in mammalian cell environment by splicing out the interfering GAL1 promotor sequence. When the resulting vector containing LacZ as a gene was introduced into yeast and mammalian cells, both cells efficiently produced .betha.-galactosidase, dimonstrating its dual host usage.

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GALOIS GROUP OF GENERALIZED INVERSE POLYNOMIAL MODULES

  • Park, Sang-Won;Jeong, Jin-Sun
    • East Asian mathematical journal
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    • v.24 no.2
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    • pp.139-144
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    • 2008
  • Given an injective envelope E of a left R-module M, there is an associative Galois group Gal($\phi$). Let R be a left noetherian ring and E be an injective envelope of M, then there is an injective envelope E[$x^{-1}$] of an inverse polynomial module M[$x^{-1}$] as a left R[x]-module and we can define an associative Galois group Gal(${\phi}[x^{-1}]$). In this paper we extend the Galois group of inverse polynomial module and can get Gal(${\phi}[x^{-s}]$), where S is a submonoid of $\mathds{N}$ (the set of all natural numbers).

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Simultaneous Dual-Enzyme Immunoassays in a Solid Phase

  • 백세환;박순재
    • Bulletin of the Korean Chemical Society
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    • v.18 no.1
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    • pp.44-50
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    • 1997
  • A method of dual-signal generation from two different enzymes was developed and utilized to simultaneously perform dual immunoassays in a single microwell. Two enzymes selected as tracers were horseradish peroxidase (HRP) and β-galactosidase (GAL). 3, 3', 5, 5'-Tetramethylbenzidine (TMB) and chlorophenolred-β-galactopyranoside (CPRG) as chromogenic substrates for the respective enzyme were used. Although the two enzymes showed their maximum activities at distinct pH conditions (pH 5.1 for HRP and 7.5 for GAL), the enzyme reactions were able to be concurrently carried out at pH 5.75 in a dual-substrate solution without signal loss. This performance was achieved by increasing TMB concentration two-fold, introducing potassium salt as activator of GAL reaction, and extending total reaction time 50%. The signal generation method was then used for dual-enzyme immunoassays to detect antibodies with co-immobilized Hepatitis C virus antigens (core and NS5) and a Hepatitis B virus antigen (PreS(2)) in a microwell. Dose-response curves of the assays revealed cooperativity between different antigen-antibody complex formation, which suggested that dual immunoassays can only be used for qualitative screening tests unless the antigens immobilized were spatially separated.