• Title/Summary/Keyword: Tet-on system

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Controlling the Gene Expression of Corynebacterium diphtheria Toxin-A Using the Tet-On System in Mouse Embryonic Stem Cells. (Mouse Embryonic Stem Cell에서 Tetracycline-Inducible System(Tet-on System)을 이용한 Corynebacterium diphtheria Toxin-A유전자의 발현 조절)

  • 박재균;임수빈;송지환
    • Microbiology and Biotechnology Letters
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    • v.32 no.1
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    • pp.11-15
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    • 2004
  • Embryonic stem (ES) cells are derived from the inner cell mass of the blastocyst-stage embryos that can be propagated indefinitely and, at the same time, can be differentiated into all the cell types that constitute the body. Current research using ES cells is mainly focused on the efficient generation of specific cell types by employing optimal differentiation conditions, which often requires the genetic manipulation of ES cells. As a way of developing an efficient system to regulate foreign gene expression in ES cells, we have inserted the gene encoding Corynebacterium diphtheria toxin-A (DTA) into an autonomously induced plasmid under positive doxycycline control ('Tet-on' system). In this study, we demonstrate that this system can lead to the cell death of mouse ES cells by the induction of DTA expression when exposed to the tetracycline derivative, doxycycline. MTT assay showed that this induction resulted in the apoptosis of ES cells.

Efficient Control of Human G-CSF Gene Expression in the Primary Culture Cell using a FIV-Tet-On Vector System (FIV-Tet-On Vector System을 이용한 hG-CSF 유전자의 효율적인 발현 조절)

  • Kwon, Mo-Sun;Koo, Bon-Chul;Kim, Te-Oan
    • Reproductive and Developmental Biology
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    • v.31 no.3
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    • pp.153-159
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    • 2007
  • In this study, using FIV-based lentivirus vector system, we tried to express hG-CSF in tetracycline-controllable manner. hG-CSF influences the proliferation, differentiation, and survival of cells in the neutrophil lineage. To enhance stability and translation of hG-CSF transcript, WPRE sequence was also introduced into FIV-Tet-On vector at downstream region of either the hG-CSF gene or the sequence encoding rtTA. Primary culture cells (CEF, chicken embryonic fibroblast; PFF, procine fetal fibroblast) infected with the recombinant FIV were cultured in the medium supplemented with or without doxycycline for 48 hours, and induction efficiency was measured by comparing the hG-CSF gene expression level using quantitative real-time PCR, Western blot and ELISA. Higher hG-CSF expression and tighter expression control were observed from the vector in which the WPRE sequence was placed at downstream of the hG-CSF (in CEF) or rtTA (in PEE) gene. This FIV-Tet-On vector system may be helpful in solving serious physiological disturbance problems which has continuously hampered successful production of transgenic animals and gene therapy.

$Hg^{2+}$-induced Aquation of trans-[Co(3,2,3-tet)$X_2]^+$ (3,2,3-tet = 4,7-diazadecane-1,10-diamine, $X_2\;=\;Cl_2,\;(NO_2)Cl,\;Br_2,\;(NO_2)Br,\;and\;(NO_3)_2)$ Complexes in Aqueous Solution (수용액에서 $Hg^{2+}$에 의한 trans-[Co(3,2,3-tet)X$_2]^+$ (3,2,3-tet = 4,7-diazadecane-1,10-diamine, $X_2\;=\;Cl_2,\;(NO_2)Cl,\;Br_2,\;(NO_2)Br,\;(NO_3)_2)$ 착물의 아쿠아 반응)

  • Doo Cheon Yoon;Chang Eon Oh;Myung Ki Doh
    • Journal of the Korean Chemical Society
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    • v.37 no.11
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    • pp.951-960
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    • 1993
  • $Hg^{2+}$-induced aquation trans-[Co(3,2,3-tet)$X_2]^+$(3,2,3-tet = 4,7-diazadecane-1,10-diamine, $X_2\;=\;Cl_2,\;(NO_2)Cl,\;Br_2,\;(NO_2)Br,\;(NO_3)_2)$ complexes was investigated in aqueous solution. The products and the reaction mechanism were confirmed by chromatography, UV/Vis. spectrum, and circular dichroism (CD) spectrum. From the results, $Hg^{2+}$-induced aquation of 3,2,3-tet system has been produced cis-${\beta}$ complex via trans complex. The kinetic studies on $Hg^{2+}$-induced aquation of trans-[Co(3,2,3-tet)$Cl_2]^+$ complex and trans-[Co(3,2,3-tet)$(NO_2)Cl]^+$ complex were also carried out to study the reaction mechanism. The results show that trans-[Co(3,2,3-tet)$Cl_2]^+$ complex undergoes the D(dissociative)-mechanism and trans-[Co(3,2,3-tet)$(NO_2)Cl]^+$ complex $I_d$(interchange dissociavite)-mechanism. In order to confirm steric course for the reaction mechanism, $Hg^{2+}$-induced aquation on trans-[Co(R,R-3,2,3-tet)$Cl_2]^+$ complex to which chiral R,R-3,2,3-tet was coordinated instead of the racemic(R,R:S,S) 3,2,3-tet was used has been examined by CD spectrum. From the results, the final complex was confirmed to be ${\Delta}-cis-{\beta}$-[Co(R,R-3,2,3-tet)$(OH_2)_2]^{3+}$ complex indicating the chirality was retained through whole process.

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Measurement and Control of the Resonance Frequency for the Transcutaneous Energy Transmission System (TET) Using the Phase Locked Loop Circuit (PLL) (PLL을 이용한 무선 전력전송 장치의 공진 주파수의 계측 및 주파수 제어)

  • Choi, S.W.;Shim, E.B.
    • Proceedings of the KSME Conference
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    • 2008.11a
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    • pp.1613-1616
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    • 2008
  • A Transcutaneous Energy Transmission System (TET) has been developed for the wireless energy transmission with two magnetically coupled coils. A resonance circuit is used to raise the induced voltage and current of the secondary coil. Its resonance frequency depends on the internal resistance of circuit and the transferred energy. Because the transferred energy usually changes in wide range, the output voltage is unstable and the energy transferring efficiency decrease. A push-pull class E amplifier is usedto generate high frequency AC voltage. To maintain proper resonance frequency, the voltage output of the amplifier was continuously monitored and adjusted to the optimized resonance frequency. Because of its high frequency (370 kHz), a phase lockedloop circuit and a comparator are used to monitor the output waveform. The results of experimentaldata show that the PLL circuit can increase the transmission efficiency and stabilize the output voltage of TET.

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Evaluation of tTA-Mediated Gene Activation System on Human Cytomegalovirus and Herpes Simplex Virus Type-1 Infections

  • Choi, Kwang-Hoon;Kim, Ki-Ho;Kim, Hong-Jin
    • Archives of Pharmacal Research
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    • v.23 no.3
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    • pp.257-260
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    • 2000
  • The tetracycline-controlled transactivator (tTA)-mediated gene activation system was examined in virus infected cells to determine its role in the control of gene expression. In the presence of tTA, the gene expression from the tetO-modified minimal promoter was efficiently activated in the uninfected cells, whereas essentially no activation was observed from the only minimal promoter without the seven direct repeats of 42 bp tetO sequences. However, essentially no activation was observed when only the minimal promoter was used, without the seven direct repetitions of the 42 bp tetO sequences. On the other hand, in the infected cells, a substantial background of $\beta$-glucuronidase expression was detected in the absence of tTA, even though tTA stimulated the gene expression by ~7-fold. This background expression indicates that the sequences within or nearby tetO are involved in the background stimulation of the gene expression by HCMV and HSV-1 . These results suggest that the application of the tTA-mediated gene activation system may not be extremely useful for studying the biological roles of HCMV and HSV genes In the viral replicative cycles, because of the basal activity of the gene expression.

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Effects of Tetrandrine and Fangchinoline on Human Platelet Aggregation, Thromboxane B$_2$ Formation and Blood coagulation.

  • Zhang, Yong-He;Kim, Hack-Seang;Yun, Yeo-Pyo;Lee, Hyung-Kyu
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1998.11a
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    • pp.177-177
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    • 1998
  • In the previous report, tetrandrine (TET) and fangchinoline (FAN) showed antithrombotic and antiplatelet aggregation activities. The present study was undertaken to investigate the effects of tetrandrine and fangchinoline on human platelet aggregation, formation of thromboxane B$_2$ and coagulation of platelet poor plasma. TET and FAN inhibited platelet activating factor (PAF) induced human platelet aggregation, but didn't inhibit the specific binding of PAF to its receptor. Meanwhile, TET and FAN also inhibited PAF, thrombin and arachidonic acid induced thromboxane B$_2$ formation in human washed platelets. In addition, neither TET nor FAN showed any anticoagulation activities in the measurement of the activated partial thromboplastin time (APTT), prothrombin time (PT) and thrombin time (TT) using human platelet poor plasma. These results suggest that antithrombotic effects of TET and FAN in mice may be mainly related to the antiplatelet aggregation activities, and the antiplatelet aggregation effects may be related to the intracellular messenger system such as TXA$_2$ formation etc., but not to the binding of PAF to PAF-receptor on the platelet membrane directly.

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TET2DICOM-GUI: Graphical User Interface Based TET2DICOM Program to Convert Tetrahedral-Mesh-Phantom to DICOM-RT Dataset

  • Se Hyung Lee;Bo-Wi Cheon;Chul Hee Min;Haegin Han;Chan Hyeong Kim;Min Cheol Han;Seonghoon Kim
    • Progress in Medical Physics
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    • v.33 no.4
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    • pp.172-179
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    • 2022
  • Recently, tetrahedral phantoms have been newly adopted as international standard mesh-type reference computational phantoms (MRCPs) by the International Commission on Radiological Protection, and a program has been developed to convert them to computational tomography images and DICOM-RT structure files for application of radiotherapy. Through this program, the use of the tetrahedral standard phantom has become available in clinical practice, but utilization has been difficult due to various library dependencies requiring a lot of time and effort for installation. To overcome this limitation, in this study a newly developed TET2DICOM-GUI, a TET2DICOM program based on a graphical user interface (GUI), was programmed using only the MATLAB language so that it can be used without additional library installation and configuration. The program runs in the same order as TET2DICOM and has been optimized to run on a personal computer in a GUI environment. A tetrahedron-based male international standard human phantom, MRCP-AM, was used to evaluate TET2DICOM-GUI. Conversion into a DICOM-RT dataset applicable in clinical practice in about one hour with a personal computer as a basis was confirmed. Also, the generated DICOM-RT dataset was confirmed to be effectively implemented in the radiotherapy planning system. The program developed in this study is expected to replace actual patient data in future studies.

Induction of cancer cell-specific death via MMP2 promoterdependent Bax expression

  • Seo, Eun-Jeong;Kim, Se-Woon;Jho, Eek-hoon
    • BMB Reports
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    • v.42 no.4
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    • pp.217-222
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    • 2009
  • Controlled gene expression in specific cells is a valuable tool for gene therapy. We attempted to determine whether the lentivirus-mediated Tet-On inducible system could be applied to cancer gene therapy. In order to select the genes that induce cancer cell death, we compared the ability of the known pro-apoptotreic genes, Bax and tBid, and a cell cycle inhibitor, p21cip1/waf1, and determined that Bax was the most effective. For the cancer cell-specific expression of $rtTA2^S$-M2, we tested the matrix metalloproteinase-2 (MMP-2) promoter and determined that it is highly expressed in cancer cell lines, including SNU475 cells. The co-transduction of two lentiviruses that contain sequences for TRE-Bax and $rtTA2^S$-M2, the expression of which is controlled by the MMP-2 promoter, resulted in the specific cell death of SNU475, whereas other cells with low MMP-2 expression did not evidence significant cell death. Our data indicate that the lentivirus-mediated Tet-On system using the cancer-specific promoter is applicable for cancer gene therapy.

A Study on the Recruitment and Selection of Secondary Science Teachers in Korea

  • Kwak, Young-Sun;Lee, Yang-Rak
    • Journal of the Korean earth science society
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    • v.31 no.5
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    • pp.488-499
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    • 2010
  • In this research, we investigated the historical development and ways of improvement of the teacher employment test (TET, hereafter) in Korea. This paper consists of three parts. The first part details the secondary science teacher education system in Korea. The second part elaborates upon the development of the TET since 1990's. The third part provides conclusion by addressing ways to improve science teacher education and employment systems in Korea. After all, the keen competition for teacher education and the demanding entry test ensure that secondary science teachers are selected from a pool of candidates with high academic achievement. Korean teacher employment system in general places more weight on subject knowledge. Although we cannot simply conclude that Korean science teachers must have profound knowledge in the subject matter and are competent in pedagogy, it stands to reason that the teachers could be more competent in performing their roles than those of many western countries with an acute shortage of teachers. We also suggested future directions and ways of improvement regarding teacher education and the TET in Korea.

Design and Implementation of Korean Tet-to-Speech System (다이폰을 이용한 한국어 문자-음성 변환 시스템의 설계 및 구현)

  • 정준구
    • Proceedings of the Acoustical Society of Korea Conference
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    • 1994.06c
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    • pp.91-94
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    • 1994
  • This paper is a study on the design and implementation of the Korean Tet-to-Speech system. In this paper, parameter symthesis method is chosen for speech symthesis method and PARCOR coeffient, one of the LPC analysis, is used as acoustic parameter, We use a diphone as synthesis unit, it include a basic naturalness of human speech. Diphone DB is consisted of 1228 PCM files. LPC synthesis method has defect that decline clearness of synthesis speech, during synthesizing unvoiced sound In this paper, we improve clearness of synthesized speech, using residual signal as ecitation signal of unvoiced sound. Besides, to improve a naturalness, we control the prosody of synthesized speech through controlling the energy and pitch pattern. Synthesis system is implemented at PC/486 and use a 70Hz-4.5KHz band pass filter for speech imput/output, amplifier and TMS320c30 DSP board.

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