• Title/Summary/Keyword: basic template

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Developing New Mammalian Gene Expression Systems Using the Infectious cDNA Molecular Clone of the Japanese Encephalitis Virus

  • Yun Sang-Im;Choi Yu-Jeong;Park Jun-Sun;Kim Seok-Yong;Lee Young-Min
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2003.05a
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    • pp.83-86
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    • 2003
  • Major advances in positive-sense RNA virus research have been facilitated by the development of reverse genetics systems. These systems consist of an infectious cDNA clone that encompasses the genome of the virus in question. This clone is then used as a template for the subsequent synthesis of infectious RNA for the generation of synthetic viruses. However, the construction of infectious cDNA for the Japanese encephalitis virus (JEV) has been repeatedly thwarted by the instability of its cDNA. As JEV is an important human pathogen that causes permanent neuropsychiatric sequelae and even fatal disease, a reliable reverse genetics system for this virus is highly desirable. The availability of this tool would greatly and the development of effective vaccines as well as facilitate studies into the basic biology of the virus, including the molecular mechanisms of viral replication, neurovirulence, and pathogenesis. We have successfully constructed a genetically stable infectious JEV cDNA containing full-length viral RNA genome. Synthetic RNA transcripts generated in vitro from the cDNA were highly infectious upon transfection into susceptible cells, and the cDNA remained stable after it had been propagated in E. coli for 180 generations. Using this infectious JEV cDNA, we have successfully expressed a variety of reporter genes from the full-length genomic and various subgenomic RNAs in vitro transcribed from functional JEV cDNAS. In summary, we have developed a reverse genetics system for JEV that will greatly facilitate the research on this virus in a variety of different fields. It will also be useful as a heterologous gene expression vector and aid the development of a vaccine against JEV.

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An Implementation of Functional Module Editor inthe Gate-Array Layout Style (게이트 어레이 레이아웃 형태에서의 기능 모듈 편집기의 구현)

  • Hong, Seong-Hyeon;Jeong, Yeong-Suk;Im, Jong-Seok;Son, Jin-U
    • The Transactions of the Korea Information Processing Society
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    • v.3 no.5
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    • pp.1240-1252
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    • 1996
  • In this paper we propose a layout editor for the functional module generation in the Sea-of Gates(SOG) lay-out style. The proposed layout editor provides interactive was of designing a functional module to the designer so that the layout result is very satisfiable. Especially, the editor is independent on the shape of the basic cells in the gate array template, and provides semi-automatic layout methods as well as hand layout. It also has several special functions which are not able to find in other layout tools for the module generation, and hence the designer can generate modules very fast. The layout editors implemented in C language with X-win-dow Motif environment. When we compare our editor with the previous layout editor Seadali, the design time is reduced by a factor of two for several benchmark circuits.

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Synthesis of Mesoporous Tin Oxide and Its Application as a Gas Sensor (메조세공을 갖는 이산화 주석의 합성 및 가스센서로서의 응용)

  • Kim, Nam-Hyon;Kim, Geon-Joong
    • Applied Chemistry for Engineering
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    • v.18 no.2
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    • pp.142-147
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    • 2007
  • In this study, mesoporous tin oxide was synthesized by sol-gel method using $C_{16}TMABr$ surfactant as a template in a basic condition. The optimum conditions for the synthesis of mesoporous $SnO_2$ were investigated and the obtained samples were characterized by XRD, nitrogen adsorption and TEM analysis. A mesoporous and nanostructured $SnO_2$ gas sensor with Au electrode and Pt heater has been fabricated on alumina substrate as one unit via a screen printing process. Sensing abilities of fabricated sensors were examined for CO and $CH_4$ gases, respectively, at $350^{\circ}C$ in the concentration range of 1~10,000 ppm. Influence of loading amount of palladium impregnated on $SnO_2$ was also tested in detection of those gases. High sensitivity to detecting gases and the fast response speed with stability were obtained with the mesoporous tin oxide sensor as compared to a non-porous one under the same detection conditions.

Identification and Characterization of Expansins from Bursaphelenchus xylophilus (Nematoda: Aphelenchoididae)

  • Lee, Dae-Weon;Seo, Jong Bok;Kang, Jae Soon;Koh, Sang-Hyun;Lee, Si-Hyeock;Koh, Young Ho
    • The Plant Pathology Journal
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    • v.28 no.4
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    • pp.409-417
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    • 2012
  • We identified two novel expansin (EXP) genes in the expressed sequence tag database of Bursaphelenchus xylophilus, designated as Bx-EXPB2 and -EXPB3. Novel Bx-EXPBs encoded 150 amino acids and their similarities in coding sequence were 70.7-84.0% to the previously reported EXPB1 of B. xylophilus. Bx-EXPB2 and Bx-EXPB3 were clustered with Bx-EXPB1 and Bm-EXPB1, respectively, forming the independent phylogeny with other nematode EXPs. All identified Bx-EXPBs contained the signal peptide and were only expressed during the propagative stage, suggesting that they are secreted to facilitate nematode migration through hosts by loosening cell walls during infection. Quantitative real-time PCR analysis showed that the relative accumulation of Bx-EXPB3 mRNAs was the highest among the three Bx-EXPs examined and the order of mRNA accumulation was as follows: Bx-EXPB3 > Bx-EXPB2 >> Bx-EXPB1. Homology modeling of Bx-EXPBs showed that the structurally optimum template was EXLX1 protein of Bacillus subtilis, whichshared residues essential for catalytic activity with Bx-EXPB1 and Bx-EXPB2 except for Bx-EXPB3. Taken together, Bx-EXPB1 and Bx-EXPB2 may be involved migration through plant tissues and play a role in pathogenesis.

Realtime Facial Expression Recognition from Video Sequences Using Optical Flow and Expression HMM (광류와 표정 HMM에 의한 동영상으로부터의 실시간 얼굴표정 인식)

  • Chun, Jun-Chul;Shin, Gi-Han
    • Journal of Internet Computing and Services
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    • v.10 no.4
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    • pp.55-70
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    • 2009
  • Vision-based Human computer interaction is an emerging field of science and industry to provide natural way to communicate with human and computer. In that sense, inferring the emotional state of the person based on the facial expression recognition is an important issue. In this paper, we present a novel approach to recognize facial expression from a sequence of input images using emotional specific HMM (Hidden Markov Model) and facial motion tracking based on optical flow. Conventionally, in the HMM which consists of basic emotional states, it is considered natural that transitions between emotions are imposed to pass through neutral state. However, in this work we propose an enhanced transition framework model which consists of transitions between each emotional state without passing through neutral state in addition to a traditional transition model. For the localization of facial features from video sequence we exploit template matching and optical flow. The facial feature displacements traced by the optical flow are used for input parameters to HMM for facial expression recognition. From the experiment, we can prove that the proposed framework can effectively recognize the facial expression in real time.

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Loop-mediated Isothermal Amplification (LAMP) for Detection of Streptococcus parauberis (Loop-mediated isothermal amplification (LAMP)법을 이용한 Streptococcus parauberis 의 신속 진단)

  • Moon, Kyung-Mi;Kim, Dong-Hwi;Heo, Moon-Soo
    • Journal of Life Science
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    • v.24 no.4
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    • pp.428-436
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    • 2014
  • Loop-mediated isothermal amplification (LAMP) technique relies on autocycling strand displacement DNA sysnthesis without template denaturation steps under isothermal conditions. LAMP has more advantages than modern PCR, as it requires only basic laboratory equipment like an isothermal water bath, oven, and heating cabinet. Hence, in this study, five random primers were designed with Streptococcus parauberis, shikimate kinase Arok gene sequences (Genbank accession number: CP0024711). Two primers were selected based on the ladder pattern. Other optimum reaction conditions like temperature, time, and sensitivity were established and confirmed with conventional SYBR-green PCR. Results confirmed that the designed random primers were species specific, without any non-target DNA amplification under isothermal conditions. Hence, this study suggests that LAMP techniques could be used in the diagnosis of fish pathogen, specifically S. parauberis.

Optimization of RAPD-FCR Conditions for Morus alba L. (뽕나무(Morus alba L.)의 RAPD 분석조건 최적화에 관한 연구)

  • 정대수;양보경;김나영;정순재;남재성;이영병;이재헌;김경태;김도훈
    • Journal of Life Science
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    • v.14 no.1
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    • pp.110-114
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    • 2004
  • The optimized RAPD-PCR conditions, that can be utilized as a basic information for analysis of the gelletic characteristics were developed for genetic analysis of four mulberry varieties, named Milsung, Chungil, Suil, and Hansung using a primer, OPY15 (5'-AGTCGCCCTT-3') from Operon company. We tested several different factors for best PCR condition including concentrations of DNA, primer, Mgclu annealing temperature, number of PCR cycle, and prosence/absence of pre-heating time at the begining of PCR reaction in the $25 \mul$volume. The best RAPD profiles were obtained using 50 ng of DNA, 1 $\mu$M of primer, $1 \mum$of $MgCl_2\;,45^{\circ}C$ of annealing temperature and an absence of pre-heating time. An establishment of the stable and reproducible RAPD-PCR conditions are expected to be useful for the subsequent RAPD-related investigation, such as genetic characterization of the mulberry varieties, re-establishment of phylogenetic relationships and development of new varieties.

Modeling Studies of an Exotype Alginate Lyase Atu3025 from Agrobacterium Tumefaciens Strain C58, a Member of Polysaccharide Lyase Family 15

  • Kothandan, Gugan;Cho, Seung Joo
    • Journal of Integrative Natural Science
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    • v.3 no.2
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    • pp.72-77
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    • 2010
  • Alginate lyases, also known as alginases or alginate depolymerases, catalyze the degradation of alginate by a ${\beta}$-elimination mechanism that has yet to be fully elucidated. Alginate is a copolymer of ${\alpha}$-L-guluronate (G) and its C5 epimer ${\beta}$-D-mannuronate (M), arranged as homopolymeric G blocks, M blocks, alternating GM or random heteropolymeric G/M stretches. Almost all alginate lyases depolymerize alginate in an endolytical fashion via a ${\beta}$-elimination reaction. The alginate lyase Atu3025 from Agrobacterium tumefaciens strain C58, consisting of 776 amino-acid residues, is a novel exotype alginate lyase classified into polysaccharide lyase family 15. Till now there is no crystal structure available for this class of proteins. Since there is no template with high sequence identity, three-dimensional coordinates for exotype alginate lyase (PL 15 family) were determined using modeling methods (Comparitive modeling and Fold recognition). The structures were modeled using the X-ray coordinates from Heparinase protein family (PDB code: 3E7J). This enzyme (Atu3025) displays enzymatic activity for both poly-M and poly-G alginate. Since poly-M is widespread; docking of a tri-mannuronate against the modeled structure was performed. We identified some of those residues which are crucial for lyase activity. The results from this study should guide future mutagenesis studies and also provides a starting point for further proceedings.

Fabrication of Hollow Micro-particles with Nonspherical Shapes by Surface Sol-gel Reaction (표면 솔-젤 반응을 활용한 마이크로미터 크기의 비구형상 공동 입자의 제조)

  • Cho, Young-Sang;Jeon, Seog-Jin;Yi, Gi-Ra
    • Korean Chemical Engineering Research
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    • v.45 no.6
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    • pp.611-618
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    • 2007
  • We demonstrate the sol-gel coating technique of colloidal clusters for producing hollow micro-particles with complex morphologies. Cross-linked amidine polystyrene (PS) microspheres were synthesized by emulsifier-free emulsion copolymerization of styrene and divinylbenzene. The amidine PS particles were self-organized inside toluene-in-water emulsion droplets to produce large quantities of colloidally stable clusters. These clusters were coated with thin silica shell by sol-gel reaction of tetraethylorthosilicate (TEOS) and ammonia, and the organic polystyrene cores were removed by calcination at high temperature to generate nonspherical hollow micro-particles with complex morphologies. This process can be used to prepare hollow particles with shapes such as doublets, tetrahedra, icosahedra, and others.

A development of a Digital tongue diagnosis system using the tongue color analysis of the each taste region (미각 영역별 설색 분석을 이용한 디지털 설진 시스템 개발)

  • Choi, Min;Yang, Dong-Min;Lee, Kyu-Won
    • Journal of the Korea Institute of Information and Communication Engineering
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    • v.19 no.2
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    • pp.428-434
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    • 2015
  • A new tongue examination model by using color analysis according to the taste division of tongue. The proposed system consists of a tongue image acquisition in a predefined template, taste region segmentation, color distribution analysis and abnormality decision of tongue by color analysis using Hue-Saturation histograms and the part of a mobile application service. We divided 4 basic taste(bitter, sweet, salty and sour) regions and performed color distribution analysis targeting each region under HSI(Hue Saturation Intensity) color model. To minimize the influence of illumination, the histograms of H and S components only except U are utilized. Using the analyzed results, the abnormality is discriminated by the criteria of the histogram range of normal tongues. Finally, a self tongue diagnosis system which can be used anytime and anywhere on mobile environment.