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Heterologous Expression and Optimized One-Step Separation of Levansucrase via Elastin-like Polypeptides Tagging System

  • Kang, Hye-Jin;Kim, Jin-Hee;Chang, Woo-Jin;Kim, Eung-Soo;Koo, Yoon-Mo
    • Journal of Microbiology and Biotechnology
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    • v.17 no.11
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    • pp.1751-1757
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    • 2007
  • Elastin-like polypeptides (ELPs) undergo a reversible inverse phase transition upon a change in temperature. This thermally triggered phase transition allows for a simple and rapid means of purifying a fusion protein. Recovery of ELPs-tagged fusion protein was easily achieved by aggregation, triggered either by raising temperature or by adding salt. In this study, levansucrase has been used as a model enzyme in the development of a simple one-step purification method using ELPs. The levansucrase gene cloned from Pseudomonas aurantiaca S-4380 was tagged with various sizes of ELPs to functionally express and optimize the purification of levansucrase. One of two ELPs, ELP[V-20] or ELP[V-40], was fused at the C-terminus of the levansucrase gene. A levansucrase-ELP fusion protein was expressed in Escherichia coli $DH5{\alpha}$ at $37^{\circ}C$ for 18 h. The molecular masses of levansucrase-ELP[V-20] and levansucrase-ELP[V-40] were determined as 56 kDa and 65 kDa, respectively. The phase transition of levansucrase-ELP[V-20] occurred at $20^{\circ}C$ in 50 mM Tris-Cl (pH 8) buffer with 3 M NaCl added, whereas the phase transition temperature ($T_t$) of levansucrase-ELP[V-40] was $17^{\circ}C$ with 2 M NaCl. Levansucrase was successfully purified using the phase transition characteristics of ELPs, with a recovery yield of higher than 80%, as verified by SDS-PAGE. The specific activity was measured spectrophotometrically to be 173 U/mg and 171 U/mg for levansucrase-ELP[V-20] and levansucrase-ELP[V-40], respectively, implying that the ELP-tagging system provides an efficient one-step separation method for protein purification.

UV Responsive Characteristics of n-Channel Schottky Barrier MOSFET with ITO as Source/Drain Contacts

  • Kim, Tae-Hyeon;Lee, Chang-Ju;Kim, Dong-Seok;Sung, Sang-Yun;Heo, Young-Woo;Lee, Jung-Hee;Hahm, Sung-Ho
    • Journal of Sensor Science and Technology
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    • v.20 no.3
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    • pp.156-161
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    • 2011
  • We fabricated a schottky barrier metal oxide semiconductor field effect transistor(SB-MOSFET) by applying indium-tin-oxide(ITO) to the source/drain on a highly resistive GaN layer grown on a silicon substrate. The MOSFET, with 10 ${\mu}M$ gate length and 100 ${\mu}M$ gate width, exhibits a threshold gate voltage of 2.7 V, and has a sub-threshold slope of 240 mV/dec taken from the $I_{DS}-V_{GS}$ characteristics at a low drain voltage of 0.05 V. The maximum drain current is 18 mA/mm and the maximum transconductance is 6 mS/mm at $V_{DS}$=3 V. We observed that the spectral photo-response characterization exhibits that the cutoff wavelength was 365 nm, and the UV/visible rejection ratio was about 130 at $V_{DS}$ = 5 V. The MOSFET-type UV detector using ITO, has a high UV photo-responsivity and so is highly applicable to the UV image sensors.

Transcriptional activation of human GM3 synthase (hST3Gal V) gene by valproic acid in ARPE-19 human retinal pigment epithelial cells

  • Song, Na-Ree;Kim, Seok-Jo;Kwon, Haw-Young;Son, Sung-Wook;Kim, Kyoung-Sook;Ahn, Hee-Bae;Lee, Young-Choon
    • BMB Reports
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    • v.44 no.6
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    • pp.405-409
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    • 2011
  • The present study demonstrated that valproic acid (VPA) transcriptionally regulates human GM3 synthase (hST3Gal V), which catalyzes ganglioside GM3 biosynthesis in ARPE-19 human retinal pigment epithelial cells. For this, we characterized the promoter region of the hST3Gal V gene. Functional analysis of the 5'-flanking region of the hST3Gal V gene revealed that the -177 to -83 region functions as the VPA-inducible promoter and that the CREB/ATF binding site at -143 is crucial for VPA-induced expression of hST3Gal V in ARPE-19 cells. In addition, the transcriptional activity of hST3Gal V induced by VPA in ARPE-19 cells was inhibited by SP600125, a c-Jun N-terminal kinase (JNK) inhibitor. In summary, our results identified the core promoter region in the hST3Gal V promoter and for the first time demonstrated that ATF2 binding to the CREB/ATF binding site at -143 is essential for transcriptional activation of hST3Gal V in VPA-induced ARPE-19 cells.

An Enhanced Greedy Message Forwarding Protocol for Increasing Reliability of Mobile Inter-Vehicle Communication (이동하는 차량 간 통신의 신뢰성 향상을 위한 개선된 탐욕 메시지 포워딩 프로토콜)

  • Ryu, Min-Woo;Cha, Si-Ho;Cho, Kuk-Hyun
    • Journal of the Institute of Electronics Engineers of Korea TC
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    • v.47 no.4
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    • pp.43-50
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    • 2010
  • Vehicle-to-Vehicle (V2V) is a special type of vehicle ad-hoc network (VANET), and known as a solution to provide communication among vehicles and reduce vehicle accidents. Geographical routing protocols as Greedy Perimeter Sateless Routing (GPSR) are very suitable for the V2V communication due to special characters of highway and device for vehicles. However, the GPSR has problem that appears local maximum by some stale neighbor nodes in the greedy mode of the GPSR. It can lose transmission data in recovery mode, even if the problem is can be solved by the recovery mode of the GPSR. We therefore propose a Greedy Perimeter Reliable Routing (GPRR), can provide more reliable data transmission, to resolve the GPSR problem in the V2V environment. Simulation results using ns-2 shown that the GPRR reveals much better performance than the GPSR by remarkably reducing the local maximum rate in the greedy mode.

Recovery Yields of Protopectinase Depending on Treatments of Organic Solvents (유기용매의 처리에 따른 Bacillus subtillis IFO 12113 유래 Protopectinase의 회수)

  • Yuk, Hyun-Gyun;Hwang, Yong-Il;Lee, Seung-Cheol
    • Applied Biological Chemistry
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    • v.40 no.2
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    • pp.107-111
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    • 1997
  • To recover protopectinase (PPase) secreted from Bacillus subtilis IFO 12113, culture filtrate of the microorganism was treated with acetone, methanol, and ethanol, respectively. In the case of treatment with acetone at a ratio of 1: 1 (culture filtrate: acetone, v/v), PPase was purified 1.7-fold with 59.2% recovery The recovery of PPase was increased by increasing the acetone concentration. PPase was purified 4-fold with 100% recovery when the culture filtrate was precipitated with methanol at a ratio of 1 : 2 (culture filtrate: methanol, v/v). However, recovery of PPase was decreased by increasing the methanol concentration. PPase was purified 13.5-fold resulting in 68% recovery by the addition of ethanol with the final ratio 1 : 1(culture filtrate: ethanol, v/v) to the supernatant, which was obtained after precipitation of the culture filtrate with ethanol at a ratio of 1 : 0.5. These results show that methanol treatment is better than other organic solvent treatments for the simple recovery of PPase, whereas fractionated treatment of ethanol can recover PPase with higher purification fold.

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Characterization of Vanadium Oxide Supported on $TiO_2-ZrO_2$ Catalysts by $^{51}V$ Solid-State NMR Spectroscopy

  • Park, Eun-Hee;Lee, Sung-Won;Lee, Man-Ho
    • Journal of the Korean Magnetic Resonance Society
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    • v.4 no.1
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    • pp.1-11
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    • 2000
  • Supported vanadium oxides are being used extensively as catalysts for a variety of reactions, including partial oxidations and ammoxidations. A series of vanadium oxide supported on TiO2-ZrO2 was obtained by impregnating ammonium metavanadate slowly into a mixed precipitateof Ti(OH)4-Zr(OH)4, followed by calcining in air at high temperatures. The prepared catalysts were characterized by 51V solid-state NMR. In the calcined catalysts 51V NMR studies indicated the peaks corresponding to distorted tetrahedral vanadia species at low V2O5 contents and octahedral vanadia species at high vanadia loadings. These results illustrate the suitability of 51V NMR as a unique quantitative spectroscopic tool in the structural analysis of vanadium(V) oxide catalytic materials.

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Development of Accessories for 345kV XLPE Cable with Small Size Conductor (345kV XLPE 소도체 Cable용 접속함의 개발)

  • Kim, H.J;Kim, K.Y;Park, J.K.;Song, J.H.;Shin, H.D.;Park, K.L.
    • Proceedings of the KIEE Conference
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    • 2004.07c
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    • pp.1775-1777
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    • 2004
  • Taihan Electric Wire Co. developed accessories(PJ, EB-G, EB-A, EB-O) for 345kV XLPE $1C{\times}600mm^2$ cable as a small size conductor cable, after installed 345kV XLPE $1C{\times}600mm^2$ cable, PJ and EB-G on Youngseo${\sim}$Youngdeungpo project KEPCO planed in 2003. All of 345kV XLPE $1C{\times}600mm^2$ cable Accessories are finished the type tests successfully. Straight joint as a Prefabricated Joint(PJ) had shorter the days of assembling than those of TMJ used on the 154kV XLPE Cable. Particularly, the use of clean booth to prevent the dusts and the penetration of water also had made the quality of jointing improved.

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Potential Characteristics of Supercapacitor Based on Ruthenium Oxide-Aqueous Electrolyte (루테늄 산화물-수계 전해액 수퍼캐패시터의 전위 특성)

  • Doh, Chil-Hoon;Choi, Sang-Jin;Moon, Seong-In;Yun, Mun-Su;Yug, Gyeong-Chang;Kim, Sang-Gil;Lee, Ju-won
    • Journal of the Korean Electrochemical Society
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    • v.6 no.2
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    • pp.93-97
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    • 2003
  • The electrode for a supercapacitor was prepared using an amorphous ruthenium oxide, which was synthesized from ruthenium trichloride hydrate$(RuO_2{\cdot}nH_2O)$. A supercapacitor was assembled with an electrode of ruthenium oxide material on a current collector of tantalum, and an electrolyte of 4.8 M sulfuric acid. The result of the AC impedance analyses on $Ta/H_2SO_4(4.8 M)/Pt$ cell showed that tantalum was stable at the potential range of $0.0\~1.1V(vs. SCE)$. Therefore, Ta film could be used the supercapacitor as a current collector. The irreversible hydrolysis in the supercapacitor occurred over ca. 1.0V(vs.SCE) when the supercapacitor was protonated to 0.5V(vs. SCE). The supercapacitor protonated to 0.5V(vs.SCE) showed good electrochemical properties when it was tested at the potential range of 1.0V in the charge-discharge test. The potential range of the electrodes including the positive and the negative electrode was varied between -0.004 and 0.995V(vs. SCE). The potential ranges of the positive and the negative electrode were $-0.004\~0.515V(vs.\;SCE)\;and\; 0.515\~0.995V(vs.\;SCE)$, respectively.

A New SOI LDMOSFET Structure with a Trench in the Drift Region for a PDP Scan Driver IC

  • Son, Won-So;Kim, Sang-Gi;Sohn, Young-Ho;Choi, Sie-Young
    • ETRI Journal
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    • v.26 no.1
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    • pp.7-13
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    • 2004
  • To improve the characteristics of breakdown voltage and specific on-resistance, we propose a new structure for a LDMOSFET for a PDP scan driver IC based on silicon-on-insulator with a trench under the gate in the drift region. The trench reduces the electric field at the silicon surface under the gate edge in the drift region when the concentration of the drift region is high, and thereby increases the breakdown voltage and reduces the specific on-resistance. The breakdown voltage and the specific on-resistance of the fabricated device is 352 V and $18.8 m{\Omega}{\cdot}cm^2$ with a threshold voltage of 1.0 V. The breakdown voltage of the device in the on-state is over 200 V and the saturation current at $V_{gs}=5V$ and $V_{ds}$=20V is 16 mA with a gate width of $150{\mu}m$.

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Studies on DC Polarograms of Some Rare Earth Elements (몇가지 희토류원소의 DC 폴라로그램에 관한 연구)

  • Cha, Ki-Won;Kim, Jea-Kyun;Kim, Sung-Il;Kim, Kyung-Whan
    • Analytical Science and Technology
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    • v.16 no.3
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    • pp.206-211
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    • 2003
  • The DC polarograms of some rare earth elements have been investigated in various pH solution and electrolytes. Samarium ion has two well-defined reduction waves at -1.75 V and -1.95 V in 0.1 M NaCl solution and those are a diffusion controlled in nature. Europium and ytterbium ions give also two step reduction waves at -0.75 V and -1.95 V for europium and -1.45 V and -2.00 V for ytterium in $(C_2H_5)_4NCl$ solution. On the other hand, lantanium and neodium ions show a single reduction wave at -1.75 V. The differences of half wave potentials between europium, ytterium and samarium ions make it possible to determine each ions in the mixed solution sponteniously. In case of europium ion, the stability constants for the complexs of $Eu^{3+}$-EDTA and $Eu^{3+}$-DTPA are evaluated.