• 제목/요약/키워드: Y2H library

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Solid-phase Parallel Synthesis of a Novel N-[Alkylsulfonamido-spiro(2H-1-benzopyran-2,4-piperidine)-6-yl] substituted Amide and Amine Drug-like Libraries

  • Kim, Ji-Hye;Gong, Young-Dae;Lee, Gee-Hyung;Seo, Jin-Soo
    • Bulletin of the Korean Chemical Society
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    • v.33 no.1
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    • pp.128-136
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    • 2012
  • We report the solid-phase library construction of 222 number of a novel N-[alkyl sulfonamido-spiro(2H-1-benzopyran-2,4-piperidine)-6-yl] substituted amide 1A and amine 1B derivatives. The polymer-bound N-[alkylsulfonamido-spiro(2H-1-benzopyran-2,4-piperidine)-6-yl] substituted amide 9 and amine 10 derivatives were obtained by first diversity generation with various acid chlorides and alkyl halides. Further reactions on the resins 9 and 10 with substituted sulfonyl chlorides produced the desired N-[alkylsulfonamido-spiro(2H-1-benzopyran-2,4-piperidine)-6-yl] substituted amide 1A and amine 1B analogues.

Construction of a High-Quality Yeast Two-Hybrid Library and Its Application in Identification of Interacting Proteins with Brn1 in Curvularia lunata

  • Gao, Jin-Xin;Jing, Jing;Yu, Chuan-Jin;Chen, Jie
    • The Plant Pathology Journal
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    • v.31 no.2
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    • pp.108-114
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    • 2015
  • Curvularia lunata is an important maize foliar fungal pathogen that distributes widely in maize growing area in China, and several key pathogenic factors have been isolated. An yeast two-hybrid (Y2H) library is a very useful platform to further unravel novel pathogenic factors in C. lunata. To construct a high-quality full length-expression cDNA library from the C. lunata for application to pathogenesis-related protein-protein interaction screening, total RNA was extracted. The SMART (Switching Mechanism At 5' end of the RNA Transcript) technique was used for cDNA synthesis. Double-stranded cDNA was ligated into the pGADT7-Rec vector with Herring Testes Carrier DNA using homologous recombination method. The ligation mixture was transformed into competent yeast AH109 cells to construct the primary cDNA library. Eventually, a high qualitative library was successfully established according to an evaluation on quality. The transformation efficiency was about $6.39{\times}10^5$ transformants/$3{\mu}g$ pGADT7-Rec. The titer of the primary cDNA library was $2.5{\times}10^8cfu/mL$. The numbers for the cDNA library was $2.46{\times}10^5$. Randomly picked clones show that the recombination rate was 88.24%. Gel electrophoresis results indicated that the fragments ranged from 0.4 kb to 3.0 kb. Melanin synthesis protein Brn1 (1,3,8-hydroxynaphthalene reductase) was used as a "bait" to test the sufficiency of the Y2H library. As a result, a cDNA clone encoding VelB protein that was known to be involved in the regulation of diverse cellular processes, including control of secondary metabolism containing melanin and toxin production in many filamentous fungi was identified. Further study on the exact role of the VelB gene is underway.

Discovery of a Novel 2,6-Difunctionalized 2H-Benzopyran Inhibitors Toward Sphingosylphosphorylcholine Synthetic Pathway as New Anti-inflammatory Target

  • Lee, Gee-Hyung;Lee, Seong Jin;Jeong, Dae Young;Kim, Ha-Young;Lee, Doohyun;Lee, Taeho;Hwang, Jong-Yeon;Park, Woo Kyu;Kong, Jae-Yang;Cho, Heeyeong;Gong, Young-Dae
    • Bulletin of the Korean Chemical Society
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    • v.35 no.8
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    • pp.2385-2390
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    • 2014
  • Novel 2,6-difuctionalized 2H-benzopyrans were synthesized and evaluated for a sphingosylphosphorylcholine(SPC) inhibitor. The synthetic 2H-benzopyrans 1c and 3a showed high potency in SPC-induced cell proliferation assay ($IC_{50}$ < 20 nM). Neither hERG $K^+$ channel binding (> $10{\mu}M$) nor CYP inhibitions (> $10{\mu}M$) were observed. Also, the simple structure-activity relationship (SAR) results were obtained from analysis of 2H-benzopyran derivatives 1-3 and the anti-SPC effect of 2H-benzopyran 1c was confirmed by a HUVEC tube formation assay.

Assembly of a Functional cDNA for Human Liver Growth Hormone Receptor: Cloning of Assembled hGHR cDNA (Human Liver로부터 Cloning한 cDNA성장호르몬 수용체의 기능성 검토)

  • 장규태;지선병홍;손동수;서원진삼;고교적웅
    • Journal of Embryo Transfer
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    • v.13 no.2
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    • pp.159-172
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    • 1998
  • 사람 성장호르몬 수용체(hGHR) cDNA는 PCR방법에 의하여 fagment로서 보고되어진 바 있으나, liver cDNA로 부터 전장을 cloning한 보고는 없는 실정으로 본 연구에서는 기능을 가진 약 4.6kbp의 cDNA hGHR을 cloning 하는데 성공하였다. 먼저 cloning하기 위하여 human liver mRNA와 human breast cancer tissue로부터 회수한 mRNA를 RT-PCR방법에 의하여 human cDNA library와 cloning에 필요한 probe를 제작하였다. human library mRNA는 GT-PCR방법에 의하여 증폭하여 증폭되어진 산물은 λZAP Vector를 이용하여 cDNA library를 구축하였고,screeing을 위하여 임 보고 되어진 hGHR fragment native sequence를 기초로 N-terminal부분의 primer를 설계하여 950bp의 probe를 얻는데 성공하였다. 이 probe를 이용하여 준비된 human liver cDNA library로부터 2.5$\times$10 6개의 plaque로부터 6개의 positive clone을 획득하였고, 이들중 poly Asignal인 "AATAAA"를 포함하고 있는 가장 긴 약 3.8kbp의 clone을 sequencing한 결과 open reading frame을 포함하고 있었으나, 5'부분의 결손되어 있었다. 그리하여 이 부분은 human breast cancer tissue로 부터 회수한 mRNA를 RT-PCR에 의하여 증폭하였고, sequencing결과 이미 보고되어진 native hGHR와 비교한 결과 하나의 nucleotide가 silent mutation으로 판명되었다.한편 human liver cDNA library로부터 cloning한 3.8cp의 positive clone의 5'end의 결손된 부분에 silent mutation된 PCR 산물을 연결함으로써 native hGHR와 유사한 cDNA hGHR subcloning에 성공하였다. 이러한 cDNA hGHR의 clone이 function을 가지고 있는지를 검토하기 위하여 eukaryotic 발현 vector인 pCXN2에 의거 ligation한 후 chinese hamster ovary cell[CHO-KI]에 transfect를 실시하였다. Dexamethasone은 첨가하지 않고 hGH만의 존재하에서 이들 cell을 배양시키고 cell menbrane에서 발현 여부를 판정키 위하여 hGHR monocloual antibody를 사용하여 flow cytometery해석을 실시하는 한편 125I-hGH binding assay에 의하여 hGH binding activity를 측정하였다. 최종적으로 GH signal transduction의 target genedf으로 알려져 있는 serine protease inhibitor 2.1(Spi 2.1) gene의 promotor activity를 검토한 결과 hGHR을 transfect한 CHO Cell에 있어서 hGH의 농도에 의존적으로 증가되었다. 따라서 본 실험에서 cloning한 cDNA hGHR는 native hGHR와 같은 기능을 가지는 것으로 판명되었다.것으로 판명되었다.

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A Meta-Ontology for Automated Information Integration of Parts Libraries (부품 라이브러리 정보의 자동 통합을 위한 메타 온톨로지)

  • Cho, J.M.;Han, S.H.;Kim, H.
    • Korean Journal of Computational Design and Engineering
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    • v.11 no.4
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    • pp.273-288
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    • 2006
  • Information integration of heterogeneous digital parts libraries or electronic pars catalogs is one of issues in B2B procurements. We need to provide an integrated view for multiple information sources. Utilization of ontologies as the metadata descriptions of the information sources can provide an integrated view. However, in order to integrate independently developed ontologies, the mismatches among them should be resolved. In this paper, we propose an ontology of meta-concepts, i.e. meta-ontology. The meta-concepts play the role of vocabulary to describe the parts library ontologies and provide well-established ontological semantics that helps the ontology modelers to consistently identify parts library concepts and systematically structure them. Consequently, the meta-ontology reduces the differences in the way the parts knowledge is interpreted and ensures the mismatches are confined to manageable mis matches, so that a software program can merge automatically. Modeling ontologies of mold and die parts libraries for B2B e-commerce is taken as an example to show how to use the meta-ontology. We also discuss how a parts library mediation system can automatically merge the well-structured parts library ontologies.

An Acquisition Policy Study by the Use Survey of a Public Library: Focused on the Analysis of Circulation Records of the H-public Library Users in 2007 (이용조사를 통한 공공도서관의 수서정책에 관한 연구 - H도서관 이용자의 2007년 대출기록을 중심으로 -)

  • Yoo, Kyeong-Jong;Park, Il-Jong
    • Journal of the Korean Society for Library and Information Science
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    • v.42 no.2
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    • pp.371-392
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    • 2008
  • The books and circulation-related data in the Online Public Access Catalog System of the H-Public library were collected and analyzed in this paper. The methods which may be applied to the Customer Relationship Management in a public library based on the results were also suggested here. Collected data were 57,927 bibliographic data of books, 11,871 patron data and 27,145 circulation records. The type of collections. circulation frequencies, total number of circulated books, publication year, and use factor as relation factor from the analyzed data of circulation records were extracted. They were also analyzed, and verified by various statistical methods such as correlation coefficient, non-parametric method, etc.

A Study of the Aging Properties of the Common Synthetic Resin Adhesives for Paper Conservation Purposes (시판용 접착제의 지류보존처리 적합성 판단을 위한 열화 특성 연구)

  • Sung, Yeonsim;Youm, Inkyung
    • Journal of Conservation Science
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    • v.28 no.4
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    • pp.387-394
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    • 2012
  • Most of the libraries and archives that lacks conservation lab facilities, frequently apply common synthetic resin adhesives such as PVAc group(Pa), acrylic group(Ac) and PVP group(Pv) on simple repairs. The archival quality synthetic adhesive(Pa-1) that is used at the National Library of Korea and the common synthetic resin adhesives were selected for the experiments. Accelerated aging test was applied on the adhesive samples and change of the pH and chromaticity were measured for one month. As the result of the pH mesurments, Pa-1 and Ac-2 were at the acceptable pH range. As the result of the chromaticity mesurments, degree of yellowing(${\Delta}b^*$) showed smaller change to greater change in the following order; Pa-1

Research Output of the Pakistani Library and Information Science Authors: A Bibliometric Evaluation of Their Impact

  • Anwar, Mumtaz Ali;Jan, Sajjad Ullah
    • Journal of Information Science Theory and Practice
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    • v.5 no.2
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    • pp.48-61
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    • 2017
  • This paper uses 601 cited papers of Pakistani LIS researchers with the purpose to examine the individual performance of these Library and Information Science (LIS) researchers in terms of their research output and its impact on the LIS (national/international) literature by using various bibliometric indicators. A list of 139 authors was compiled with the help of the Library, Information Science, and Technology Abstracts (LISTA) and some other sources. Data were collected from Google Scholar and SPSS version 20 was utilized in order to identify the relationship between self-citations and various performance indices of the authors. The average citations received per paper vary from 1.80 to 10.08. About half of the papers were single-authored whereas less than one-fifth were by three or more authors. The authors who worked in collaboration produced more papers and received more citations. The h-index, g-index, hI-index, hI-norm, and e-index were used to determine the rank for each author. The intra-group citations grid revealed the volume of self-citations and a small group who cite each other more due to close academic and social relationships. The correlations between self-citations and the impact indices used revealed significant differences. Findings are useful for concerned institutions regarding award, promotions, etc. Further, future research should seriously consider the self-citations and social networking of authors while examining their citations-based research performance.