• Title/Summary/Keyword: linked library

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Isolation and Characterization of Single-Chain Fv Against Ductal Cells

  • Lee Myung-Hoon;Ryu Hye-Myung;Kim Sun-Zoo;Park Ji-Young;Uhm Ji-Hyun;Park Tae-In
    • Biomedical Science Letters
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    • v.10 no.3
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    • pp.211-217
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    • 2004
  • For discrimination of ductal and ascinar cells, we isolated a single-chain variable domain fragment (scFv) antibody against ductal cells of salivary gland using phage display technique. From the spleen of a mouse immunized with ductal cell lysate, total RNA was prepared and used as a template for cDNA synthesis of antibody genes. The scFv genes were constructed with variable domain genes of heavy and light chain and were introduced into pCANTAB5E to construct phage scFv library. The phage particles specific for acinar cells were screened by subtraction using immunotubes coated with acinar and ductal cell lysate and enzyme-linked immunoabsorbance assay (ELISA). The characteristics of the scFv were determined by immunohistochemistry (IHC) and the result indicated that the isolated scFv has the specificity against ductal cells of salivary glands and tubules of kidney. And the scFv has an unique binding activity specific for Hashimoto's thyroiditis. The nucleotide sequence of isolated scFv gene was determined and revealed that V/sub H/ belongs to the mouse H-chain family subgroup IB and V/sub L/ to the mouse L-chain family subgroup III.

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Genome Research on Peach and Pear

  • Hayashi Tateki;Yamamoto Toshiya
    • Proceedings of the Korean Society of Plant Biotechnology Conference
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    • 2002.04a
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    • pp.101-109
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    • 2002
  • A lot of SSRs (simple sequence repeats) in peach and pear from enriched genomic libraries and in peach from a cDNA library were developed. These SSRs were applied to other related species, giving phenograms of 52 Prunus and 60 pear accessions. Apple SSRs could also be successfully used in Pyrus spp. Thirteen morphological traits were characterized on the basis of the linkage map obtained from an $F_2$ population of peach. This map was compiled with those morphological markers and 83 DNA markers, including SSR markers used as anchor loci, to compare different peach maps. Molecular markers tightly linked to new root-knot nematode resistance genes were also found. A linkage map including disease-related genes, pear scab resistance and black spot susceptibility, in the Japanese pear Kinchaku were constructed using 118 RAPD markers. Another linkage map, of the European pear Bartlett, was also constructed with 226 markers, including 49 SSRs from pear, apple, peach and chewy. Maps of other Japanese pear cultivars, i.e., Kousui and Housui, were also constructed. These maps were the first results of pear species.

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Web-Based Electronic Design Automation Circuit Design and Part Management System Development (WEB 통합 Electronic Design Automation 회로설계 및 부품 관리 시스템 개발에 관한 연구)

  • Kang, Do-Young;Hha, Ki-Jong;Choi, Young-Gyu
    • Journal of Advanced Navigation Technology
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    • v.12 no.3
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    • pp.255-262
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    • 2008
  • In this paper, we developed the circuit and part information maintenance system based on JSP (Java Server Page) web server using web browser. Designers can easily search part information and register the new components or parts, and this system is linked to the EDA (Electronic Design Automation) system to be applicable to the circuit and part schematics in real time. Therefore, EDA system part library managed by individual designers are integrated into the web-based maintenance system.

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Development Strategy for functional rice improved with human lactoferrin and enhancement of nutrient compounds (인체 모유 단백질 및 영양 성분 강화 고부가가치 기능성 쌀 생산 벼 품종 개발 전략)

  • Rhim Seong-Lyul;Lee Jin-Hyoung;Lee Hyo-Yeon;Suh Suk-Cheol
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2002.05a
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    • pp.48-50
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    • 2002
  • A strategy for development of a functional rice in proved with human lactoferrin and enhancement of nutrient compounds was planned. For the purposes we have cloned and characterized a human lactoferrin cDNA from human mammary gland cDNA library A endosperm storage vacuole targeting sequence and the cDNA fragment was linked to endosperm specific glutelin promoter. The fusion gene fragment was inserted into a binary vector containing MAR gene. In addition a new ${\beta}$-galactosidase gene from Bifidobacterium of human was used as a reporter gene in the vector system, Rice plants showing a high concentration of amino acids in the endosperm cells were developed by using a biochemical mutation and bred for the transformation with the binary vector system Finally we have established a transformation method for the rice endosperm cells.

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Screening of Peptides Bound to Anthrax Protective Antigen by Phage Display

  • Kim, Joung-Mok;Park, Hye-Yeon;Choi, Kyoung-Jae;Jung, Hoe-Il;Han, Sung-Hwan;Lee, Jae-Seong;Park, Joon-Shik;Yoon, Moon-Young
    • Journal of Microbiology and Biotechnology
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    • v.16 no.11
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    • pp.1784-1790
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    • 2006
  • Bacillus anthracis is a causative agent of anthrax. Anthrax toxins are composed of a protective antigen (PA), lethal factor (LF), and edema factor (EF), in which the PA is a central mediator for the delivery of the two enzymatic moieties LF and EF. Therefore, the PA has been an attractive target in the prevention and vaccinization for anthrax toxin. Recently, it has been reported that the molecule consisting of multiple copies of PA-binding peptide, covalently linked to a flexible polymer backbone, blocked intoxification of anthrax toxin in an animal model. In the present study, we have screened novel diverse peptides that bind to PA with a high affinity (picomolar range) from an M13 peptide display library and characterized the binding regions of the peptides. Our works provide a basis to develop novel potent inhibitors or diagnostic probes with a diverse polyvalence.

Expression and Characterization of Trehalose Biosynthetic Modules in the Adjacent Locus of the Salbostatin Gene Cluster

  • Choeng, Yong-Hoon;Yang, Ji-Yeon;Delcroix, Gaetan;Kim, Yoon-Jung;Chang, Yong-Keun;Hong, Soon-Kwang
    • Journal of Microbiology and Biotechnology
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    • v.17 no.10
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    • pp.1675-1681
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    • 2007
  • The pseudodisaccharide salbostatin, which consists of valienamine linked to 2-amino-1,5-anhydro-2-deoxyglucitol, is a strong trehalase inhibitor. From our Streptomyces albus ATCC 21838 genomic library, we identified thirty-two ORFs in a 37-kb gene cluster. Twenty-one genes are supposed to be a complete set of modules responsible for the salbostatin biosynthesis. Through sequence analysis of the gene cluster, some of the upstream gene products (SalB, SalC, SalD, SalE, and SalF) revealed functional resemblance with trehalose biosynthetic enzymes. On the basis of this rationale, we isolated the five genes (salB, salC, salD, salE, and salF) from the S. albus ATCC 21838 and cloned them into the expression vector pWHM3. We demonstrated the noticeable expression and accumulation of trehalose, using only the five upstream biosynthetic gene cluster of salbostatin, in the transformed Streptomyces lividans TK24. Finally, 490 mg/l trehalose was produced by fermentation of the transformant with sucrosedepleted R2YE media.

Molecular Characterization of AceB, a Gene Encoding Malate Synthase in Corynebacterium glutamicum

  • Lee, Heung-Shick;Anthony J. Sinskey
    • Journal of Microbiology and Biotechnology
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    • v.4 no.4
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    • pp.256-263
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    • 1994
  • The aceB gene, encoding for malate synthase, one of the key enzymes of glyoxylate bypass, was isolated from a pMT1-based Corynebacterium glutamicum gene library via complementation of an Escherichia coli aceB mutant on an acetate minimal medium. The aceB gene was closely linked to aceA, separated by 598 base pairs, and transcribed in divergent direction. The aceB expressed a protein product of Mr 83, 000 in Corynebacterium glutamicum which was unusually large compared with those of other malate synthases. A DNA-sequence analysis of the cloned DNA identified an open-reading frame of 2, 217 base pairs which encodes a protein with the molecular weight of 82, 311 comprising 739 aminoo acids. The putative protein product showed only limited amino acid-sequence homology to its counteliparts in other organisms. The N-terminal region of the protein, which shows no apparent homology with the known sequences of other malate synthases, appeared to be responsible for the protein s unusually large size. A potential calciumbinding domain of EF-hand structure found among eukaryotes was detected in the N-terminal region of the deduced protein.

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Genome Research on Peach and Pear

  • Hayashi, Tateki;Yamamoto, Toshiya
    • Proceedings of the Korean Society of Plant Biotechnology Conference
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    • 2002.04b
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    • pp.101-109
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    • 2002
  • A lot of SSRs (simple sequence repeats) in peach and pear from enriched genomic libraries and in peach from a cDHA library were developed. These SSRs were applied to other related species, giving phenograms of 52 Prunus and 60 pear accessions. Apple SSRs could also be successfully used in Pyrus spp. Thirteen morphological traits were characterized on the basis of the linkage map obtained from an Fa population of peach. This map was compiled with those morphological markers and 83 DHA markers, including SSR markers used as anchor loci, to compare different peach maps. Molecular markers tightly linked to new root-knot nematode resistance genes were also found. A linkage map including disease-related genes, pear scab resistance and black spot susceptibility, in the Japanese pear Kinchaku were constructed using 118 RAPD markers. Another linkage map, of the European pear Bartlett, was also constructed with 226 markers, including 49 SSRs from pear, apple, peach and cherry. Maps of other Japanese pear cultivars, i.e., Kousui and Housui, were also constructed. These maps were the first results of pear species.

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Neutralizing Chimeric Mouse-human Antibodies against Burkholderia pseudomallei Protease: Expression, Purification and Characterization

  • Chan, Shzu-Wei;Ong, Guan-Im;Nathan, Sheila
    • BMB Reports
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    • v.37 no.5
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    • pp.556-564
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    • 2004
  • A recombinant Fab monoclonal antibody (Fab) C37, previously obtained by phage display and biopanning of a random antibody fragment library against Burkholderia pseudomallei protease, was expressed in different strains of Escherichia coli. E. coli strain HB2151 was deemed a more suitable host for Fab expression than other E. coli strains when grown in media supplemented with 0.2% glycerol. The expressed Fab fragment was purified by affinity chromatography on a Protein G-Sepharose column, and the specificity of the recombinant Fab C37 towards B. pseudomallei protease was proven by Western blotting, enzyme-linked immunosorbent assay (ELISA) and by proteolytic activity neutralization. In addition, polyclonal antibodies against B. pseudomallei protease were produced in rabbits immunized with the protease. These were isolated from high titer serum by affinity chromatography on recombinant-Protein A-Sepharose. Purified polyclonal antibody specificity towards B. pseudomallei protease was proven by Western blotting and ELISA.

Characteristics and Industrial Linkage of Science Parks : A Case Study of the United Kingdom (영국 과학단지의 특성과 기업연계)

  • 조혜영
    • Journal of the Korean Regional Science Association
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    • v.15 no.1
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    • pp.45-61
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    • 1999
  • A rapid growth of science parks worldwide since 1980s is caused by both the widespread perception that technological innovation is the essential element for the economic development and the notified cases of successful high technology cluster like Silicon Valley. Analyzed in terms of R & D employment and expenditure, new product launches, patent registration, qualifications of the founder, self-rated technological level and the like, the technological level of the science parks in the U.K. reveals very high. While nealy half of the firms on the science parks are represented to be formally linked with universities, the use of the library is referred most frequently among the types of the linkages. R & D links such as contract research, sponsored research, test and analysis were rarely mentioned. Summing up, science parks play an important role for the development of high-tech industries with superior technological characteristics. But in reatlity, there remains so much difference among the individual parks and their location. In addition the individual parks and their location. In addition, institutionalization of the technology transfer with univeristies that only when the science parks are based on the innovative environment they can be successful in the capability of the technological innovation.

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