• BMB Reports
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• v.33 no.2
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• pp.166-171
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• 2000

Thioredoxin is a multifunctional protein that is ubiquitous in microorganisms, animals and plants. Previously, the expression of the Escherichia coli thioredoxin gene (trxA) was found to be negatively regulated by cAMP. In the present study, the effect of temperature on the expression of the E. coli trxA gene was investigated. In order to examine the temperature effect, the fusion plasmid pCL70 that harbors the E. coli trxA P1P2 promoter was used. The other two fusion plasmids, pJH3 and pMH521 that were constructed in different vectors which harbor the E. coli trxA P2 promoter, were also used. When the E. coli strain MC1061/pCL70 was grown in a rich medium at $25^{\circ}C$, $34^{\circ}C$ and $42^{\circ}C$, the cells grown at $42^{\circ}C$ gave the highest $\beta$-galactosidase activity. The E. coli MC1061/pJH3 and MC1061/pMG521 cells showed increased $\beta$-galactosidase activity after the shift of the culture temperature to $42^{\circ}C$. The wild-type trxA gene of the E. coli MC1061 cells produced much higher thioredoxin activity at the higher temperature. These results support the conclusion that the E. coli trxA gene is regulated in a temperature-dependent manner. Especially the expression from its P2 promoter appeared to be sensitive to temperature.

• Journal of Microbiology and Biotechnology
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• v.19 no.11
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• pp.1456-1463
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• 2009

Vibrio parahaemolyticus possessing urease-positive property is relatively rare, but such strains consistently exhibit the TDH-related hemolysin (TRH) gene. In this study, we examined the effects of incubation temperature on urease activity expression, using the TH3996 and AQ4673 strains where the enzyme activity is known to be temperature-dependent and -independent, respectively. In the TH3996 strain, $\beta$-galactosidase activity was 4.4-fold lower after $30^{\circ}C$ cultivation than after $37^{\circ}C$ in a ureR-lacZ fusion strain, but temperature dependency was not found in ureD- or nikA-lacZ fusion strains. However, ureR-, ureD-, and nikA-lacZ fusions of the AQ4673 strain was not influenced by incubation temperature. We compared the promoter sequences of ureR between the above two strains. Intriguingly, we detected mismatches of two nucleotides between the two strains located at positions -66 and -108 upstream of the methionine initiation codon for UreR. Additionally, urease activity was not affected by culture temperature at either $30^{\circ}C$ or $37^{\circ}C$ by allelic introduction of the AQ4673 ureR gene into the TH3996 ureR deletion mutant. Taken together, our study demonstrates that the transcriptional factor UreR is involved in the temperature dependency of urease activity, and two nucleotides within the ureR promoter region are of particular importance for the urease activity dependency of V. parahaemolyticus.

• Journal of the Korean Vacuum Society
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• v.16 no.6
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• pp.483-488
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• 2007

Current feeder system (CFS) for Korea superconducting tokamak advanced research(KSTAR) project plays a role to interconnect magnet power supply (MPS) and superconducting (SC) magnets through the normal bus-bar at the room temperature(300 K) environment and the SC bus-line at the low temperature (4.5 K) environment. It is divided by two systems, i.e., toroidal field system which operates at 35 kA DC currents and poloidal field system wherein 20$\sim$26 kA pulsed currents are applied during 350 s transient time. Aside from the vacuum system of main cryostat, an independent vacuum system was constructed for the CFS in which a roughing system is consisted by a rotary and a mechanical booster pump and a high vacuum system is developed by four cryo-pumps with one dry pump as a backing pump. A self interlock and its control system, and a supervisory interlock and its control system are also established for the operational reliability as well. The entire CFS was completely tested including the reliability of local/supervisory control/interlock, helium gas leakage, vacuum pressure, and so on.

• International Journal of Aeronautical and Space Sciences
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• v.18 no.2
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• pp.206-214
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• 2017

In this study, measurement of thermophysical properties of materials at high temperatures was performed. This experiment employed a heater device to heat the material to a high temperature. The images of the specimen surface due to thermal load at various temperatures were recorded using charge-coupled device (CCD) cameras. Afterwards, the full-field thermal deformation of the specimen was determined using the digital image correlation (DIC) method. The capability and accuracy of the proposed technique are verified by two experiments: (1) thermal deformation and strain measurement of a stainless steel specimen that was heated to $590^{\circ}C$ and (2) thermal expansion and thermal contraction measurements of specimen in the process of heating and cooling. This research focused on two goals: first, obtaining the temperature dependence of the coefficient of thermal expansion, which can be used as data input for finite element simulation; and second, investigating the capability of the DIC method in measuring full-field thermal deformation and strain. The results of the measured coefficient of thermal expansion were close to the values available in the handbook. The measurement results were in good agreement with finite element method simulation results. The results reveal that DIC is an effective and accurate technique for measuring full-field high-temperature thermal strain in engineering fields such as aerospace engineering.

• 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.

• Transactions of the Korean hydrogen and new energy society
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• v.24 no.1
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• pp.36-43
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• 2013

In this paper, a three-dimensional hydrogen desorption model is applied to a thin double-layered annulus ZrCo hydride bed and validated against the temperature evolution data measured by Kang et al. The present model reasonably captures the bed temperature evolution behavior and the 90% hydrogen discharging time. In addition, the performance of thin double-layered annulus bed is evaluated by comparing with a simple cylindrical bed using hydrogen desorption model. This study provides multi-dimensional contours such as temperature and H/M atomic ratio in the metal hydride region. This numerical study provides fundamental understanding during hydrogen desorption process and indicates that efficient design of the metal hydride bed is critical to achieve rapid hydrogen discharging performance. The present three-dimensional hydrogen desorption model is a useful tool for the optimization of bed design and operating conditions.

• Nuclear Engineering and Technology
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• v.41 no.5
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• pp.709-714
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• 2009

An ion implanter, which can serve as a metal-ion supply, has been constructed and performance tested. Copper ions are generated and extracted from a Bernas ion source with a heating crucible that provides feed gases to sustain the plasma. Sable arc plasmas can be sustained in the ion source for a crucible temperature in excess of $350^{\circ}C$. Stable extraction of the ions is possible for arc Currents less than 0.3 A. Arc currents increase with the induced power of a block cathode and the transverse field in the ion source. $Cu^+$ ions in the extracted beam are separated using a dipole magnet. A $20{\mu}A$ $Cu^+$ ion current can be extracted with a 0.2 A arc current. The ion current can support a dose of $10^{16}ions/cm^2$ over an area of $15\;cm^2$ within a few hours.

• Journal of Welding and Joining
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• v.14 no.5
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• pp.122-133
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• 1996

In order to evaluate the cryogenic fracture characteristics of structural steels for superconducting magnets of fusion reactor, small punch (SP) testing was performed on austenitic stainless steel (JN1 base metal) and its TIG weldments at 293K, 77K and 4K. The mechanical properties with respect to the extracted location of the weld metal, on the effects of welding heat cycle about base metal near fusion line in TIG weldments were investigated. The mechanical property of the weld metal in TIG weldments depends on distance from welding root, root region of weldments having the lowest mechanical property. The base metal near fusion line showed degradation of mechanical property caused by cyclic heating during the TIG welding. Based on the test results, HAZ was found to be up to 5mm from the fusion line. It is shown that SP testing is a useful tool to evaluate the mechanical properties with respect to the microstructures changes such as HAZ as well as weld metal in TIG weldments at cryogenic temperature.

• Journal of Microbiology
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• v.33 no.2
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• pp.120-125
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• 1995

.betha.-galactosidase activity of Escherichia coli cells containing operon fusion nhaA'-'lacZ was monitored to study the regulation of expression of nhaA gene under various conditions. The expression of the fusion was enhanced only by chemicals containing Na$^{+}$ or Li$^{+}$. This Na$^{+}$ or Li$^{+}$. This Na$^{+}$(Li$^{+}$)-specific enhancement of .betha.-galactosidase activity represented the increase in the rate of synthesis of .betha.-galactosidase rather than the decrease in the breakdown rate. The induction pattern was influenced by copy numbers of the gene. Induction by Na$^{+}$ or Li$^{+}$ was concentration and time dependent, reaching maximum 5-6 fold induction after 2 hours at 0.4-0.5 M for Na$^{+}$ or at 0.25-0.35 M for Li$^{+}$, Although the expression was induced at much lower concentration of Na$^{+}$ at alkaline pH values than at neutral pH in the presence of Na$^{+}$, alkaline pH itself did ot induced the expression of the fusion in the absence of Na$^{+}$. Temperature shift and growth phase of culture did not affect the level of induction.he level of induction.

• BMB Reports
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• v.44 no.4
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• pp.279-284
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• 2011

The glutathione S-transferase (GST) system is useful for increasing protein solubility and purifying soluble GST fusion proteins. However, purifying half of the GST fusion proteins is still difficult, because they are virtually insoluble under non-denaturing conditions. To optimize a simple and rapid purification condition for GST-pyruvate kinase muscle 2 (GST-PKM2) protein, we used 1% sarkosyl for lysis and a 1 : 200 ratio of sarkosyl to Triton X-100 (S-T) for purification. We purified the GST-PKM2 protein with a high yield, approximately 5 mg/L culture, which was 33 times higher than that prepared using a conventional method. Notably, the GST-high-temperature requirement A2 (GST-HtrA2) protein, used as a model protein for functional activity, fully maintained its proteolytic activity, even when purified under our S-T condition. This method may be useful to apply to other biologically important proteins that become highly insoluble in the prokaryotic expression system.