• BMB Reports
• /
• v.35 no.2
• /
• pp.206-212
• /
• 2002

The NS2B-NS3(pro) polyprotein segment from the dengue virus serotype 2 strain 16681 was purified from overexpressing E. coli by metal chelate affinity chromatography and gel filtration. Enzymatic activity of the refolded NS2B-NS3(pro) protease complex was determined in vitro with dansyl-labeled peptide substrates, based upon native dengue virus type 2 cleavage sites. The 12mer substrate peptides and the cleavage products could be separated by reversed-phase HPLC, and were identified by UV and fluorescence detection. All of the peptide substrates (representing the DEN polyprotein junction sequences at the NS2A/NS2B, NS2B/NS3, NS3/NS4A and NS4B/NS5 sites) were cleaved by the recombinant protease NS2B-NS3(pro). No cleavage was observed with an enzymatically inactive S135A mutant of the NS3 protein, or with a modified substrate peptide of the NS3/NS4A polyprotein site that contained a K2093A substitution. Enzymatic activity was dependent on the salt concentration. A 50% decrease of activity was observed in the presence of 0.1M sodium chloride. Our results show that the NS3 protease activity of the refolded NS2B-NS3(pro) protein can be assayed in vitro with high specificity by using cleavage-junction derived peptide substrates.

• BMB Reports
• /
• v.33 no.4
• /
• pp.294-299
• /
• 2000

Proteolytic processing of the dengue virus serotype 2 polyprotein precursor is catalyzed by a host signal peptidase and a virus encoded two-component protease consisting of the nonstructural proteins, NS2B and NS3. We expressed in Escherichia coli the NS2B, NS3(pro) and NS2B-NS3 proteins from the dengue virus type 2 strain 16681 as N-terminal fusions with a hexahistidine affinity tag under the control of the inducible trc promoter. All fusion proteins were purified to >90% purity by detergent extraction of inclusion bodies and a single step metal chelate chromatography. Proteins were refolded on-column and recovered with yields of 0.5, 6.0 and 1.0 mg/l of E. coli culture that was grown to $OD_{600}=1.0$ for NS2B, NS3(pro) and NS2B-NS3, respectively. Purified proteins gave strong signals in Western blots using $Ni^{2+}-nitrilotriacetic$ acid as a probe for the presence of the polyHis tag. During the purification process, $(His)_{6}NS2B-NS3$ was apparently not autoproteolytically cleaved at the NS2B/NS3 site.

• Journal of Microbiology and Biotechnology
• /
• v.30 no.11
• /
• pp.1651-1658
• /
• 2020

Since Zika virus (ZIKV) was first detected in Uganda in 1947, serious outbreaks have occurred globally in Yap Island, French Polynesia and Brazil. Even though the number of infections and spread of ZIKV have risen sharply, the pathogenesis and replication mechanisms of ZIKV have not been well studied. ZIKV, a recently highlighted Flavivirus, is a mosquito-borne emerging virus causing microcephaly and the Guillain-Barre syndrome in fetuses and adults, respectively. ZIKV polyprotein consists of three structural proteins named C, prM and E and seven nonstructural proteins named NS1, NS2A, NS2B, NS3, NS4A, NS4B, and NS5 in an 11-kb single-stranded positive sense RNA genome. The function of individual ZIKV genes on the host innate immune response has barely been studied. In this study, we investigated the modulations of the NF-κB promoter activity induced by the MDA5/RIG-I signaling pathway. According to our results, two nonstructural proteins, NS2A and NS4A, dramatically suppressed the NF-κB promoter activity by inhibiting signaling factors involved in the MDA5/RIG-I signaling pathway. Interestingly, NS2A suppressed all components of MDA5/RIG-I signaling pathway, but NS4A inhibited most signaling molecules, except IKKε and IRF3-5D. In addition, both NS2A and NS4A downregulated MDA5-induced NF-κB promoter activity in a dosedependent manner. Taken together, our results suggest that NS2A and NS4A signifcantly antagonize MDA5/RIG-I-mediated NF-κB production, and these proteins seem to be controlled by different mechanisms. This study could help understand the mechanisms of how ZIKV controls innate immune responses and may also assist in the development of ZIKV-specific therapeutics.

• Journal of Korean Society for Quality Management
• /
• v.43 no.4
• /
• pp.489-498
• /
• 2015

Purpose: This paper proposes S-NS diagram. S-NS diagram helps us find core factors which improve customer satisfaction. Methods: S-NS diagram draws a scatter diagram based on marks obtained from surveys consisting of several questions on satisfaction and non-satisfaction at the same time. S-NS diagram is divided into the 4 regions. We focused on region B which is High satisfaction and high non-satisfaction shown in S-NS diagram. Region B is the areas for needs to improve. Results: S-NS diagram can find few important factors which affect customer satisfaction. We improve Lecture Satisfaction using S-NS Diagram. Conclusion: S-NS diagram has been proven to be an effective method for improving customer satisfaction with a lecture satisfaction improvement example. We know S-NS diagram can use many fields including manufacture and service areas.

• Journal of Environmental Science International
• /
• v.24 no.1
• /
• pp.55-64
• /
• 2015

The dry weight of mother plants' leaves had the highest increase rate in both NS (single-use) and NS+EM (mixed-use) mixed with NS 0.8 (customary use). In seafood amino acid fertilizer (SAF) application, the increase rate was highest in SAF solution at a 300-fold dilution. Mother plants' crown diameter, plant height, leaf length, leaf width, petiole length and leaf number showed the greatest growth amount when NS 0.8 (customary use) was mixed to NS (single-use) or NS+EM (mixed-use) solution. The growth was highest in SAF solution diluted 300 folds, but lowest in SAF solution diluted 100 folds. Of all inorganic nutrients, excluding sulfur, total amount of nitrogen, available phosphorus, potassium, calcium and magnesium had the highest increase rate in both NS (single-use) and NS+EM (mixed-use) with the treatment of NS 0.8 (customary use). Total nitrogen, in particular, was increased by 3.1% in NS 0.4, 6.0% in NS 0.8, and 4.5% in NS 0.8 with the application of NS+EM at a 500-fold dilution compared to NS alone. Total nitrogen amount showed the highest increase rate in SAF solution diluted 300 folds. Total nitrogen, available phosphorus, calcium, magnesium and EC in soils applied with culture solutions (NS, NS+EM) had increasing tendencies after fertilizer application. The results were comparable to those of SAF treatment. The increase rate of each inorganic nutrient composition declined in soils applied with NS+EM solution diluted 500 folds compared to NS alone.

• Bulletin of the Korean Chemical Society
• /
• v.20 no.3
• /
• pp.301-306
• /
• 1999

Hepatitis C virus (HCV) has been known to be an enveloped virus with a positive strand RNA genome and the major agent of the vast majority of transfusion associated cases of hepatitis. For viral replication, HCV structural proteins are first processed by host cell signal peptidases and NS2/NS3 site of the nonstructural protein is cleaved by a zinc-dependent protease NS2 with N-terminal NS3. The four remaining junctions are cleaved by a separate NS3 protease. The solution conformations of NS4B/5A, NS5A/5B substrates and NS5A/5B inhibitor have been determined by two-dimensional nuclear magnetic resonance (NMR) spectroscopy. NMR data suggested that the both NS5A/5B substrate and inhibitor appeared to have a folded tum-like conformation not only between P1 and P6 position but also C-terminal region, whereas the NS4B/5A substrate exhibited mostly extended conformation. In addition, we have found that the conformation of the NS5A/5B inhibitor slightly differs from that of NS5A/5B substrate peptide, suggesting different binding mode for NS3 protease. These findings will be of importance for designing efficient inhibitor to suppress HCV processing.

• The Bulletin of The Korean Astronomical Society
• /
• v.40 no.1
• /
• pp.88.4-89
• /
• 2015

The Double Pulsar (PSR J0737-3039) is the only neutron star-neutron star (NS-NS) binary in which both NSs have been detectable as radio pulsars. The Double Pulsar has been assumed to dominate the Galactic NS-NS binary merger rate $R_g$ among all known systems, solely based on the properties of the first-born, recycled pulsar (PSR J0737-3039A, or A) with an assumption for the beaming correction factor of 6. In this work, we carefully correct observational biases for the second-born, non-recycled pulsar (PSR J0737-0737B, or B) and estimate the contribution from the Double Pulsar on $R_g$ using constraints available from both A and B. Observational constraints from the B pulsar favour a small beaming correction factor for A (~2), which is consistent with a bipolar model. Considering known NS-NS binaries with the best observational constraints, including both A and B, we obtain $R_g=21_{-14}{^+28}$ per Myr at 95 per cent confidence from our reference model. We expect the detection rate of gravitational waves from NS-NS inspirals for the advanced ground-based gravitational-wave detectors is to be $8_{-5}{^+10}$ per yr at 95 per cent confidence. We discuss prospects of gravitational-wave detection based on our results. Implications of PSR J1906+0746, which is likely to be another tight NS-NS binary in the Galactic disc supported by recent observation, are also remarked.

• Korean Journal of Ichthyology
• /
• v.22 no.1
• /
• pp.49-55
• /
• 2010

Sympatric sites of type NE and NS of Zacco korean us were found in the upper reaches of many tributaries of the Nakdong and Seomjin rivers, as well as in some streams draining into the South Sea, Korea. In streams where the two types exist together, there was a tendency for individuals of NE to densely populate the upper reaches and those of NS to populate the middle reaches. A habitat-segregation phenomenon was observed in the cohabiting sites. Fishes of NE preferred rapids, whereas fishes of NS preferred slowly flowing or lentic regions. It seemed that the number of NE was proportional to the flow velocity, but NS showed an opposite trend in the cohabiting sites. On the other hand, such tendency was not found in regions where NE and NS exist separately.

• Journal of Korea Water Resources Association
• /
• v.41 no.7
• /
• pp.747-759
• /
• 2008

This study, a point rainfall process model, which could represent appropriately observed rainfall data, was to select. The point process models-rectangular pulses Poisson process model(RPPM), Neyman-Scott rectangular pulses Poisson process model(NS-RPPM), and modified Neyman-Scott rectangular pulses Poisson process model(modified NS-RPPM)-all based on Poisson process were considered as possible rainfall models, whose statistical analyses were performed with their simulation rainfall data. As results, simulated rainfall data using the NS-RPPM and the modified NS-RPPM represent appropriately statistics of observed data for several aggregation levels. Also, simulated rainfall data using the modified NS-RPPM shows similar characteristics of rainfall occurrence to the observed rainfall data. Especially, the modified NS-RPPM reproduces high-intensity rainfall events that contribute largely to occurrence of natural harzard such as flood and landslides most similarly. Also, the modified NS-RPPM shows the best results with respect to the total rainfall amount, duration, and inter-event time. In conclusions, the modified NS-RPPM was found to be the most appropriate model for the long-term simulation of rainfall.

• Applied Chemistry for Engineering
• /
• v.25 no.4
• /
• pp.396-400
• /
• 2014

The pore surface of mesoporous materials, SBA-15 and MCM-41 were functionalized with organosilanes, 3-aminopropyltrimethoxysilane (1NS) and N-[(3-trimethoxysilyl)propyl]ethylenediamine (2NS) via grafting method. $(n-BuCp)_2ZrCl_2$ and methylaluminoxane (MAO) were impregnated on the surface-functionalized mesoporous materials for the application to ethylene polymerization. In the case of SBA-15/2NS/$(n-BuCp)_2ZrCl_2$ supported Zr and Al contents decreased as grafted 2NS content increased. However, in the case of MCM-41/2NS/$(n-BuCp)_2ZrCl_2$ supported Al content decreased, but Zr content increased as grafted 2NS content increased. The polymerization activity of SBA-15/2NS/$(n-BuCp)_2ZrCl_2$ increased as the amount of grafted 2NS increased. Increase in the amount of grafted 2NS should caused decrease in pore volume and diameter. Consequently, it decreased the amount of supported metallocene and MAO in general. However, the smaller pore-sized MCM-41 could have lower supported MAO content due to its large molecular size in case that MCM-41 was surface-functionalized with 2NS. Therefore, the supported metallocene content could increase and its polymerization activity was higher than that of SBA-15.