• Bulletin of the Korean Chemical Society
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• 제26권2호
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• pp.285-291
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• 2005

The nonstructural protein 5B (NS5B) of hepatitis C virus (HCV) is the viral RNA-dependent RNA polymerase (RdRp), which is the essential catalytic enzyme for the viral replication and is an appealing target for the development of new therapeutic agents against HCV infection. A small amount of serum from a single patient with hepatitis C was used to get the genome of a Korean HCV isolate. Sequence analysis of NS5B 1701 nucleotides showed the genotype of a Korean isolate to be subtype 1b. The soluble recombinant HCV NS5B polymerase lacking the C-terminal 24 amino acids was expressed and purified to homogeneity. With the highly purified NS5B protein, we established in vitro systems for RdRp activity to identify potential polymerase inhibitors. The rhodanine family compounds were found to be potent and specific inhibitors of NS5B from high throughput screening (HTS) assay utilizing the scintillation proximity assay (SPA) system. The binding mode of an inhibitor was analyzed by measuring various kinetic parameters. Lineweaver-Burk plots of the inhibitor suggested it binds not to the active site of NS5B polymerase, but to an allosteric site of the enzyme. The activity of NS5B in in vitro polymerase reactions with homopolymeric RNA requires interaction with multiple substrates that include a template/primer and ribonucleotide triphosphate. Steady-state kinetic parameter, such as Km, was determined for the ribonucleotide triphosphate. One of compounds found interacts directly with the viral polymerase and inhibits RNA synthesis in a manner noncompetitively with respect to UTP. Furthermore, we also investigated the ability of the compound to inhibit NS5B-directed viral RNA replication using the Huh7 cell-based HCV replicon system. The investigation is potentially very useful for the utility of such compounds as anti-hepatitic agents.

• Asian-Australasian Journal of Animal Sciences
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• 제32권2호
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• pp.290-296
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• 2019

Objective: Pigs share many physiological, anatomical and genomic similarities with humans, which make them suitable models for biomedical researches. Understanding the genetic status of Yucatan miniature pigs (YMPs) and their association with human diseases will help to assess their potential as biomedical model animals. This study was performed to identify non-synonymous single nucleotide polymorphisms (nsSNPs) in selective sweep regions of the genome of YMPs and present the genetic nsSNP distributions that are potentially associated with disease occurrence in humans. Methods: nsSNPs in whole genome resequencing data from 12 YMPs were identified and annotated to predict their possible effects on protein function. Sorting intolerant from tolerant (SIFT) and polymorphism phenotyping v2 analyses were used, and gene ontology (GO) network and Kyoto encyclopedia of genes and genomes (KEGG) pathway analyses were performed. Results: The results showed that 8,462 genes, encompassing 72,067 nsSNPs were identified, and 118 nsSNPs in 46 genes were predicted as deleterious. GO network analysis classified 13 genes into 5 GO terms (p<0.05) that were associated with kidney development and metabolic processes. Seven genes encompassing nsSNPs were classified into the term associated with Alzheimer's disease by referencing the genetic association database. The KEGG pathway analysis identified only one significantly enriched pathway (p<0.05), hsa04080: Neuroactive ligand-receptor interaction, among the transcripts. Conclusion: The number of deleterious nsSNPs in YMPs was identified and then these variants-containing genes in YMPs data were adopted as the putative human diseases-related genes. The results revealed that many genes encompassing nsSNPs in YMPs were related to the various human genes which are potentially associated with kidney development and metabolic processes as well as human disease occurrence.

• Applied Biological Chemistry
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• 제40권1호
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• pp.48-52
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• 1997

다양한 가공 적성을 갖는 변성 전분을 토마토 케찹의 제조에 첨가하여 변성 전분의 종류 및 첨가량이 토마토 케찹의 유동 특성, 층분리 및 관능적 성질에 미치는 영향에 관하여 조사하였다. 케찹을 Herschel-Bulkley계 유체로 간주하여 분석한 결과 항복응력와 점조도지수는 ADA(acetylate distarch adipate)>SA(starch acetate)>HDP(hydroxypropyl distarch phosphate)>RCS(raw corn starch)>NS(no starch)의 순으로 크게 나타났으며 거동유동지수는 SA와 HDP의 경우 첨가량에 관계없이 거의 일정하였으나 ADA와 RCS는 첨가량이 증가할 수록 감소하는 경향을 보였다. 원심분리에 의한 층분리 검사에서는 ADA>SA>HDP>RCS>NS의 순으로 층분리가 방지되는 안정성을 나타냈다. 관능검사 결과 전반적으로 ADA(2%)>SA(2%)>RCS(2%)>NS>HDP(2%)의 순으로 기호도가 높게 나타났다.

• Journal of Microbiology and Biotechnology
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• 제18권2호
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• pp.328-333
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• 2008

A series of pep tides binding to the HCV NS5B polymerase was selected from phage display peptide libraries. A conserved motif of Ser-Arg-X-Arg/Leu was identified among the selected peptides, and Pep2 (Trp-Ser-Arg-Pro-Arg-Ser-Leu) was chosen for further characterization. The binding of Pep2 to HCV NS5B in vivo was shown by a yeast two-hybrid assay and by subcellular colocalization analysis using immunofluorescence confocal microscopy. The in vitro interaction was also confirmed by GST pulldown assay. The replication of the HCV 1b subgenomic replicon was efficiently inhibited by the presence of the peptide. By using a subtractive biopanning against Pep2, the binding site of the peptide was mapped at the pocket of Pro388 to Pro391 in the thumb subdomain of the polymerase. A yeast two-hybrid analysis using Pro388Ala and Pro391Ala mutants of NS5B confirmed the binding.

• 한국미생물·생명공학회지
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• 제36권4호
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• pp.353-359
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• 2008

Hepatitis C virus (HCV) is known as the causative agent of blood transmitted hepatitis. Two viral proteins, E2 and NS5A, are known to exert interferon resistance of HCV via PKR pathway. Here, we report a third protein, the RNA-dependent RNA polymerase (NS5B) of HCV, induced interferon resistance inhibiting p56 pathway. p56 was shown to interact with p48 subunit of eukaryotic initiation factor 3 (eIF3). This interaction inhibited formation of ternary complex in translation initiation. Using dual reporter assay system, we observed that the translation decreased when interferon alpha was added to the culture. But, in the presence of HCV NS5B, the translation partly recovered. NS5B and p48 subunit of eIF3 were shown to interact. This interaction seems to inhibit the interaction between p48 and p56. This is the first report that a virus exerts interferon resistance via p56 pathway.

• 대한전자공학회논문지TC
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• 제37권1호
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• pp.88-97
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• 2000

초고속이면서 소요 메모리의 크기를 극소화한 IP 라우터용 Lookup 알고리즘을 제안하고 성능을 분석하였다. 메모리 크기가 작으므로 고속/고가의 SRAM(10ns)을 사용할 수 있고, 구조가 간단하여 하드웨어로 구현 가능하였다. 본 장치는 1${\sim}$3회의 메모리 접근을 통해 Lookup이 가능하고, IPMA 사이트에서 구한 40,000개의 라우팅 정보를 이용하여 시뮬레이션한 결과 대략 ${\sim}$316KB의 포워딩 테이블용 메모리만이 소요된다. 이때 압축을 수행하는 옵셋 임계치는 8이다. ALTERA EPM7256시리즈에 100MHz 클럭을 이용하여 모사시험한 결과 10ns 접근속도를 가진 SRAM 기준으로 2회의 메모리 접근만으로 Lookup하는 경우 45ns의 접근시간이 소요되며, 3회의 메모리 접근이 필요한 경우는 ${\sim}$177ns의 접근시간이 소요된다.

• Molecules and Cells
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• 제21권3호
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• pp.330-336
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• 2006

Since its identification in 1989, hepatitis C virus has been the subject of extensive research. The biology of the virus and the development of antiviral drugs are closely related. The RNA polymerase activity of nonstructural protein 5B was first demonstrated in 1996. NS5B is believed to localize to the perinuclear region, forming a replicase complex with other viral proteins. It has a typical polymerase structure with thumb, palm, and finger domains encircling the active site. A de novo replication initiation mechanism has been suggested. To date, many small molecule inhibitors are known including nucleoside analogues, non-nucleoside analogues, and pyrophosphate mimics. NS5B interacts with other viral proteins such as core, NS3, 4A, 4B, and 5A. The helicase activity of NS3 seems necessary for RNA strand unwinding during replication, with other nonstructural proteins performing modulatory roles. Cellular proteins interacting with NS5B include VAMP-associated proteins, heIF4AII, hPLIC1, nucleolin, PRK2, ${\alpha}$-actinin, and p68 helicase. The interactions of NS5B with these proteins might play roles in cellular trafficking, signal transduction, and RNA polymerization, as well as the regulation of replication/translation processes.

• 화약ㆍ발파
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• 제31권1호
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• pp.55-63
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• 2013

싱가포르 전력구 터널 건설공사는 싱가포르 내의 전력수요 증가에 대응하기 위해 지하에 400kV의 고압 전기 케이블 등 송전설비 설치를 위한 터널을 건설하는 공사이다. 본 전력구는 총연장 35km의 터널로서 18.5km의 North-South Line의 3개 공구 (NS1, NS2, NS3)와 16.5km의 East-West Line의 3개 공구 (EW1, EW2, EW3)로 나누어 건설된다. 총 6개의 공구 중 SK건설은 EW2 공구와 NS2 공구를 수주하여 현재 시공중이다. 본 프로젝트의 과업 중 지상과 고압 송전 케이블 터널을 연결하는 수직구가 공구당 3~4개소가 있으며, 시공 중에는 TBM 발진용으로 활용된다. 지하 전력구는 싱가포르 내 도심 한복판을 가로질러 건설되며, 수직구 또한 대부분 도심지 내에 있어 수직구 굴착 시 발파 효율의 극대화와 동시에 싱가포르 소음 및 진동 기준을 만족하는 최적의 발파 설계가 요구된다. 싱가포르 전력구 터널 EW2 공구의 수직구 발파는 현지 허용 진동속도기준을 준수하고 국내의 우수한 발파 설계기술을 적용하여 설계되었으며 본 설계를 통하여 국내의 우수한 발파 설계 및 시공 기술을 전파할 좋은 기회가 될 것으로 기대된다.

• 정보과학회지
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• 제25권11호
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• pp.73-78
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• 2007

• 한국산림과학회지
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• 제106권1호
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• pp.19-25
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• 2017

본 연구는 몽골 반건조지에서 질소 시비가 Populus sibirica 묘목의 생리적 특성 및 생장에 미치는 영향을 파악하고자 수행되었다. 2015년 5월, 2년생 P. sibirica 묘목을 식재하고 처리구별로 묘목 당 요소 5 g (N1), 15 g (N2), 30 g (N3) 및 N2와 같은 질소량인 황산암모늄 33 g (NS) 등을 시비하였다. 잎의 엽록소 함량은 6월과 7월에 처리구 간 유의한 차이를 보이지 않았다. 그러나 엽록소 함량이 8월에는 처리구 간 차이를 보였으며 6월과 7월에 비하여 낮게 나타났는데, 이는 잎의 노화에 의한 것으로 판단된다. 6월과 7월의 순광합성률은 N2와 NS 처리구에서 대조구보다 높게 나타났다. 6월의 증산 속도는 순광합성률의 경향과 달리 N2 처리구에서 NS 처리구보다 유의하게 높게 나타났다. 한편, 묘목의 근원경상대생장률은 NS 처리구에서만 대조구보다 유의하게 높았고, 수고 상대생장률은 처리구간 유의한 차이를 보이지 않았다. 질소 시비에 따른 엽면적 차이는 처리구 간에 유의성을 보였으나, 대조구와 각 질소 처리구 간에는 유의한 차이가 없었다. 본 연구지에서 P. sibirica 묘목의 초기 생육 증진을 위하여 요소보다 황산암모늄을 시비하는 것이 더 적합한 것으로 보인다. 요소와 황산암모늄 시비가 토양 및 묘목에 미치는 영향이 다르므로 몽골에서 황산암모늄의 적정 시비량 산정을 위한 추가 연구가 필요한 것으로 판단된다.