• BMB Reports
• /
• 제35권2호
• /
• 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
• /
• 제33권4호
• /
• 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
• /
• 제30권11호
• /
• 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.

• 품질경영학회지
• /
• 제43권4호
• /
• 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.

• 한국환경과학회지
• /
• 제24권1호
• /
• 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
• /
• 제20권3호
• /
• 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.

• 천문학회보
• /
• 제40권1호
• /
• 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.

• 한국어류학회지
• /
• 제22권1호
• /
• pp.49-55
• /
• 2010

참갈겨니 NE와 NS형의 서식범위 내에서 두 형이 공서하는 지점을 조사한 결과 낙동강과 섬진강의 각 지류의 상류지점, 남해로 흐르는 일부 하천에서 발견되었다. 두 형이 공서하는 하천에서의 분포양상은 NE형은 상류 지역에 NS형은 중류 지역에 치우치는 경향을 보였다. 중간 지역에서는 같은 지점에서 동시에 출현하였으나, NE형은 여울에 NS형은 웅덩이 또는 흐름이 느린 곳에서 주로 서식하는 서식처 분리현상을 나타내었다. 유속과 출현개체수의 관계는 공서지에서는 유속의 증가에 따라 NE형은 개체수가 증가하는 반면 NS형은 감소하는 경향을 보였으나, 각각 단독으로 존재하는 곳에서는 그러한 경향이 나타나지 않았다.

• 한국수자원학회논문집
• /
• 제41권7호
• /
• pp.747-759
• /
• 2008

본 연구에서는 관측강우의 통계특성 및 발생특성을 가장 적절하게 재현해 주는 강우모형을 선정하고자 하였다. 강우모형으로 Poisson과정에 근거한 점과정모형인 RPPM, NS-RPPM, modified NS-RPPM을 고려하여 모의자료에 대한 통계분석을 수행하였다. 그 결과, NS-RPPM과 modified NS-RPPM을 이용하여 모의된 자료가 여러 집성시간의 통계치를 적절하게 재현하였다. 또한 modified NS-RPPM을 이용하여 모의된 자료가 관측자료와 가장 유사한 발생특성을 가지는 것을 알 수 있었다. 특히, 홍수, 산사태 등 자연재해의 발생에 큰 영향을 주는 큰 강도를 가지는 강우를 관측치와 가장 유사하게 재현하였다. 모의된 강우사상의 총 강우량, 강우기간, 강우사상 간의 간격을 관측강우와 비교해본 결과 또한 modified NS-RPPM이 가장 좋은 결과를 보였다. 본 연구의 결과를 종합해 볼 때, 강우의 장기 모의를 위해 modified NS-RPPM을 이용하는 것이 가장 적절할 것으로 판단된다.

• 공업화학
• /
• 제25권4호
• /
• pp.396-400
• /
• 2014

메조포러스 물질, SBA-15와 MCM-41의 세공 표면을 유기실란인 3-aminopropyltrimethoxysilane (1NS)와 N-[(3-trimethoxysilyl) propyl]ethylenediamine (2NS)를 이용하여 그라프팅법으로 표면 기능화하였다. 표면 기능화된 메조포러스 물질에 $(n-BuCp)_2ZrCl_2$와 조촉매 methylaluminoxane (MAO)를 담지하여 에틸렌 중합을 실시하였다. SBA-15/2NS/$(n-BuCp)_2ZrCl_2$ 촉매는 그라프팅되는 2NS의 양이 증가할수록 담지된 Zr와 Al 함량은 감소하였으며, MCM-41/2NS/(n-BuCp)2ZrCl2 촉매는 2NS의 양이 증가할수록 Zr 함량은 증가하지만 Al 함량은 감소하였다. SBA-15/2NS/$(n-BuCp)_2ZrCl_2$ 촉매의 중합 활성은 2NS의 투입량을 증가할수록 감소하였다. 이는 많은 양의 2NS가 표면에 그라프팅되는 경우 기공부피와 기공크기가 작아지게 되고 따라서 MAO와 메탈로센 촉매 담지량이 감소하게 되기 때문이다. 그러나 SBA-15보다 작은 기공을 갖는 MCM-41을 2NS로 표면 기능화되면 큰 분자 크기를 갖는 조촉매 MAO는 담지 되기 어려워 낮은 MAO 담지량을 갖게 된다. 따라서 메탈로센 담지량은 약간 증가하게 되고 이에 따라 중합 활성이 증가하였다.