• BMB Reports
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• 제35권2호
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• pp.206-212
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• 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
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• 제33권4호
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• pp.294-299
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• 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.

• Bulletin of the Korean Chemical Society
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• 제20권3호
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• pp.301-306
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• 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.

• Journal of Nutrition and Health
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• 제39권3호
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• pp.299-306
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• 2006

This study was done to investigate the use of nutritional supplements (NS) in Korean elite soccer players using an anonymous questionnaire. NS were classified into health supplements, manufactured health food supplements, chinese medicines & tonic foods, and nutrient supplements. Information was sought on the type of NS and factors that might influence supplement use including selected demographic parameters and health related variables. The data were collected from 241 athletes (aged $24.6{\pm}3.8$ years) in 9 professional soccer club. The prevalence of NS use among the subjects was 81.3%. Health supplements were used most frequently and nutrient supplement drug was the second one. Among the health supplements, weight/muscle gainer and calorie replacement product were most frequently used. Vitamin supplements were most frequently used among all nutrient supplement drugs. Users of NS were higher age (p<0.05) duration of exercise (p<0.05), and income (p<0.05) than non-users. The married (p<0.05) and a member of K-league (p<0.01) tended to have higher prevalence of NS use. The main adviser of NS were family (55.3%) and oneself (39%) instead of coach or sports nutritionist. Although NS use, only 26.2% certainly check up nutritional information on their used NS. Among the subjects, 84.5% of them felt that NS use were improved athletic performance, and 86.5% of them will to keep on taking supplement. These data suggest that a large number of elite soccer players use NS and these players may require education about healthy nutritional supplement practice and on the proper use of nutritional supplements.

• The Plant Pathology Journal
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• 제27권4호
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• pp.310-314
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• 2011

Nonstructural protein 3 (NS3) encoded by RNA3 of Rice stripe virus (RSV), known to be a suppressor of gene silencing, was cloned and sequenced. The cloned NS3 gene is composed of 636 nucleotides encoding 211 deduced amino acids, and showed a high degree of similarity with the equivalent genes isolated from Korea, Japan and China. The NS3 gene promoted the enhancement of transient gene expression and suppressed transgene co-silencing. In the transient GFP expression via agroinfiltration, GFP expression was dramatically enhanced in terms of both protein yield and expression period in the presence of NS3. The highest accumulation of GFP protein reached to 6.8% of total soluble proteins, which corresponded to a two-fold increase compared to that obtained in the absence of NS3. In addition, NS3 significantly suppressed the initiation of GFP co-silencing induced by the additive GFP infiltration in GFP-transgenic Nicotiana benthamiana. The NS3 gene was also found to be a stronger suppressor than Cucumber mosaic virus 2b. These observations are believed to be derived from the strong suppressive effect of NS3 on gene silencing, and indicate that NS3 could be used as an effective enhancer for the rapid production of foreign proteins in plants.

• 한국정보통신학회논문지
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• 제10권12호
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• pp.2202-2211
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• 2006

EXT3NS 파일 시스템은 Network-Storage Card(NS 카드)라는 전용 하드웨어를 기반으로 하는 멀티미디어 스트리밍 서버를 위한 파일 시스템이다. EXT3NS는 NS 카드 상의 PCI memory(PMEM)를 이용, 스트리밍 시 디스크에 있는 데이터를 메인 메모리를 거치지 않고 바로 네트워크 카드로 내보냄으로써 스트리밍 서버의 성능을 향상시킨다. 본 논문에서는 NS 카드에 있는 PMEM을 기반으로 한 버퍼 캐시를 추가 설계, 구현하고, 이 버퍼 캐시가 EXT3NS 파일 시스템을 채용한 서버의 멀티미디어 스트리밍의 성능을 개선시킬 수 있음을 보인다. 또한, 제안된 버퍼 캐시 상에서 다수의 동시 사용자를 지원하는 멀티 미디어 스트리 밍 서버를 위한 버퍼 캐시 교체 정책인 Old New Section(ONS) 교체 기법을 제안한다. 제안된 OSN 교체 기법은 멀티미디어 스트리밍 환경에서 기존의 교체 정책들 보다 좋은 성능을 보인다. 성능 평가 결과, 다시 읽기동작과 무작위 읽기 동작에서 기존의 EXT3NS 파일 시스템 보다 PMEM에 캐시를 사용한 EXT3NS가 각각 평균 33MB/sec, 2.4MB/sec의 성능이 향상 되었다. 또한, 다시 읽기 동작의 경우 기존의 Least Frequently Used(LFU) 교체 정 책을 사용한 경우보다 ONS 교체 정책을 사용한 경우가 약 600KB/sec의 성능 증가를 보인다. 이러한 결과는 동시에 여러명의 사용자 요구를 처리해야 하는 대용량 멀티 미디어 스트리밍 서버에서 보다 효과적인 읽기 동작을 처리할 수 있음을 나타낸다.

• Journal of Microbiology and Biotechnology
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• 제30권11호
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• pp.1651-1658
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• 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.

• 공업화학
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• 제25권4호
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• pp.396-400
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• 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.

• 한국균학회지
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• 제44권3호
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• pp.138-144
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• 2016

대전광역시 연자산과 충청남도 주요 산림 토양들로부터 분리, 동정한 야생효모들 중 국내 미기록종으로 Pseudozyma prolifica HL9-1, Trichosporon coremiiforme NS19-2, Candida cretensis SA4-1, Cryptococcus diffluens TJ4-3, Cryptococcus pinus YB17-2 등의 유포자효모와 Candida vartiovaarae DD2-5, Pichia galeiformis DM3-5, Candida pseudolambica JW2-3, Trichosporon xylopini NS5-1, Trichosporon moniliiforme NS5-7, Tetrapisispora iriomotensis NS14-2, Tetrapisispora nanseiensis SA17-1 등의 무포자효모들을 선별하여 이들의 미생물학적 특성을 조사하였다. 12균주 모두 구형~타원형이었고 출아법으로 영양증식하였으며 yeast extract peptone dextrose (YPD) 배지에서 잘 생육하였다. 특히 Candida cretensis SA4-1과 Tetrapisispara iriomotensis NS14-2은 10% NaCl을 함유한 YPD 배지에서 잘 생육하는 호염성 효모들이었다. Candida vartiovaarae DD2-5외 3균주들은 유당을 자화시켰으며 Tetrapisispora nanseiensis SA17-1은 xylose를 자화시키고 동시에 발효시켰다. Candida cretensis SA4-1과 Tetrapisispara iriomotensis NS14-2 두 호염성 효모들의 생리기능성을 조사한 결과 이들의 무세포추출물들의 항고혈압성 안지오텐신 전환효소 저해활성이 각각 71.3%와 68.4%로 높았다.

• 대한전기학회논문지:전기물성ㆍ응용부문C
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• 제52권10호
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• pp.476-480
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• 2003

In this paper, we have performed 3-dimensional time-resolving measurement of the Ne light emitted from the cell of plasma display panel(PDP) as a function of the pressure using the scanned point detecting system. From the temporal behavior results, we could analyze the discharge behavior of panel with Ne-Xe(4%) mixing gas and 300 torr, 400 torr and 500 torr pressure. At the top view of panel, the discharge of 300 torr panel starts at the 634 ns and ends at the 722 ns. The emission duration time is about 90 ns. The discharge of 400 torr panel starts at the 682 ns and ends at the 786 ns. the emission duration time is about 100 ns. Also, the discharge of 500 torr panel starts at the 770 ns and ends at the 826 ns. the emission duration time is about 56 ns. The discharge moves from inner edge of cathode electrode to outer cathode electrode forming arc type. In the side view of 300 torr, 400 torr and 500 torr an emission shows that the light is detected up to 180${\mu}{\textrm}{m}$, 150${\mu}{\textrm}{m}$ and 70${\mu}{\textrm}{m}$ height of barrier rib and the emission distribution of the 300 torr is wider than 400 torr, 500 torr.