• Title/Summary/Keyword: Fusion process

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The development of anti-DR4 single-chain Fv (ScFv) antibody fused to Escherichia coli alkaline phosphatase (대장균의 alkaline phosphatase가 융합된 anti-DR4 single-chain Fv (ScFv) 항체의 개발)

  • Han, Seung Hee;Kim, Jin-Kyoo
    • Korean Journal of Microbiology
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    • v.52 no.1
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    • pp.10-17
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    • 2016
  • Enzyme immunoassay to analyze specific binding activity of antibody to antigen uses horseradish peroxidase (HRP) or alkaline phosphatase (AP). Chemical methods are usually used for coupling of these enzymes to antibody, which is complicated and random cross-linking process. As results, it causes decreases or loss of functional activity of either antibody or enzyme. In addition, most enzyme assays use secondary antibody to detect antigen binding activity of primary antibody. Enzymes coupled to secondary antibody provide a binding signal by substrate-based color development, suggesting secondary antibody is required in enzyme immunoassay. Additional incubation time for binding of secondary antibody should also be necessary. More importantly, non-specific binding activity caused by secondary antibody should also be eliminated. In this study, we cloned AP isolated from Escherichia coli (E. coli) chromosome by PCR and fused to) hAY4 single-chain variable domain fragment (ScFv) specific to death receptor (DR4) which is a receptor for tumor necrosis factor ${\alpha}$ related apoptosis induced ligand (TRAIL). hAY4 ScFv-AP expressed in E. coli showed 73.8 kDa as a monomer in SDS-PAGE. However, this fusion protein shown in size-exclusion chromatography (SEC) exhibited 147.6 kDa as a dimer confirming that natural dimerization of AP by non-covalent association induced ScFv-AP dimerization. In several immunoassay such as ELISA, Western blot and immunocytochemistry, it showed antigen binding activity by color development of substrates catalyzed by AP directly fused to primary hAY4 ScFv without secondary antibody. In summary, hAY4 ScFv-AP fusion protein was successfully purified as a soluble dimeric form in E. coli and showed antigen binding activity in several immunoassays without addition of secondary antibody which sometimes causes time-consuming, expensive and non-specific false binding.

Enhancement of the solubility of human tissue inhibitor of matrix metallocroteinase-2 (TIMP-2) in E. coli using a modified in vitro mutagenesis (새로운 유전자 재조합 방법을 이용한 대장균에서의 인간 tissue inhibitor of mtrix metalloproteinase-2 (TIMP-2) 유전자의 가용성 발현)

  • Kim, Jong-Uk;Choi, Dong-Soon;Joo, Hyun;Min, Churl-K.
    • KSBB Journal
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    • v.23 no.3
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    • pp.231-238
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    • 2008
  • The second family member of tissue inhibitors of matrix metalloproteinases, TIMP-2, is a 21kDa protein which inhibits matrix metalloproteinases 2 (MMP-2). Expression of mammalian proteins in E. coli often forms inclusion bodies that are made up of mis-folded or insoluble protein aggregates. The requirement for the formation of 6 disulfide bonds in the process of the TIMP-2 folding is likely to be incompatible with the reducing environment of E. coli. However, this incompatibility can be often overcome by introducing a mutagenesis that could lead to enhancement of the protein solubility. In this reason, we have attempted to express the soluble TIMP-2 in E. coli by applying a modified staggered extension process (StEP), one of the in vitro PCR-based recombinant mutagenesis methods, and error-prone PCR. C-terminally located CAT fusion protein with respect to mutated TIMP-2 proteins enables us to differentiate the soluble TIMP-2 from the insoluble in E. coli by virtue of chloramphenicol resistance. According to this scheme, E. coli harboring properly-folded CAT fused to TIMP-2 protein was selected, and some of the resulting colonies exhibited an enhanced, soluble expression of TIMP-2 compared to the wild type, implying (i) the StEP technique is successfully employed to enhance the proper folding thereby increasing the solubility of TIMP-2, and (ii) the CAT dependent screening may be a simple and effective method to differentiate the soluble protein expression in E. coli.

Retrieval of High Resolution Surface Net Radiation for Urban Area Using Satellite and CFD Model Data Fusion (위성 및 CFD모델 자료의 융합을 통한 도시지역에서의 고해상도 지표 순복사 산출)

  • Kim, Honghee;Lee, Darae;Choi, Sungwon;Jin, Donghyun;Her, Morang;Kim, Jajin;Hong, Jinkyu;Hong, Je-Woo;Lee, Keunmin;Han, Kyung-Soo
    • Korean Journal of Remote Sensing
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    • v.34 no.2_1
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    • pp.295-300
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    • 2018
  • Net radiation is the total amount of radiation energy used as a heat flux for the Earth's energy cycle, and net radiation from the surface is an important factor in areas such as hydrology, climate, meteorological studies and agriculture. It is very important to monitoring the net radiation through remote sensing to be able to understand the trend of heat island and urbanization phenomenon. However, net radiation estimation using only remote sensing data is generally causes difference in accuracy depending on cloud. Therefore, in this paper, we retrieved and monitored high resolution surface net radiation at 1 hour interval in Eunpyeong New Town where urbanization using Communication, Ocean and Meteorological Satellite (COMS), Landsat-8 satellite and Computational Fluid Dynamics (CFD) model data reflecting the difference in building height. We compared the observed and estimated net radiation at the flux tower. As a result, estimated net radiation was similar trend to the observed net radiation as a whole and it had the accuracy of RMSE $54.29Wm^{-2}$ and Bias $27.42Wm^{-2}$. In addition, the calculated net radiation showed well the meteorological conditions such as precipitation, and showed the characteristics of net radiation for the vegetation and artificial area in the spatial distribution.

The Study on the Role of 3D Animated Pre-visualization in VFX FilmProduction (VFX 영화 제작을 위한 3D animatied Pre-visualization(3D애니메이티드 사전시각화)의 역할에 관한 연구)

  • Park, Sung-Ho
    • Cartoon and Animation Studies
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    • s.51
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    • pp.293-319
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    • 2018
  • Thanks to the advancement of the related technologies and equipment, today's video contents like movies, animations and soap operas are rapidly expanding their expressible cinematic imagination area. In order to fulfill the elevated visual expectations of audiences and realize exciting storytelling and fantastic world, the fusion of different techniques is actively used, and the reality for visual effects and image synthesis is increasing more and more. Accordingly, recent VFX-oriented movies using CG have a much more complicated production process than before. Therefore, the importance of Pre-visualization, aka Pre-vis is becoming bigger in the planning process for sophisticated design. Pre-vis means that the advance visualization for stories or directing ideas in the planning process before starting production of movies or animations. 3D animated Pre-visualization realizing directors' abstract and ambiguous ideas in 3 dimensional environment in advance is, as a powerful means for visual storytelling, briskly used focusing on the VFX film industry on which the present CG is broadly used, and the role of Pre-vis throughout productions has increased compared to the past. The studies, however, on the role and utility of Pre-vis are not enough. Therefore, this study was conducted on the role of Pre-vis used for present VFX movie productions using the examples of 3D animated Pre-visualization production in which the researcher of this study participated. In this study, the role of the Pre-vis that is subdivided presently, is divided into and 3D animatics and their each role is analyzed with the example images. Through this, the characteristics that Pre-vis should have are clarified and the concept of the advantages and utility led by the use of Pre-vis in productions is strengthened. The goal of this study is to induce active uses of Pre-vis throughout productions after forming consensus about the various roles of Pre-vis and their utility.

Analyses of the Setup Errors using on Board Imager (OBI) (On Board Imager (OBI)를 이용한 Setup Error 분석에 대한 연구)

  • Kim, Jong-Deok;Lee, Haeng-O;You, Jae-Man;Ji, Dong-Hwa;Song, Ju-Young
    • The Journal of Korean Society for Radiation Therapy
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    • v.19 no.1
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    • pp.1-5
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    • 2007
  • Purpose: The accuracy and advantages of OBI(On Board Imager) against the conventional method like film and EPID for the setup error correction were evaluated with the analysis of the accumulated data which were produced in the process of setup error correction using OBI. Materials and Methods: The results of setup error correction using OBI system were analyzed for the 130 patients who had been planned for 3 dimensional conformal radiation therapy during March 2006 and May 2006. Two kilo voltage images acquired in the orthogonal direction were fused and compared with reference setup images. The setup errors in the direction of vertical, lateral, longitudinal axis were recorded and calculated the distance from the isocenter. The corrected setup error were analyzed according to the lesion and the degree of shift variations. Results: There was no setup error in the 41.5% of total analyzed patients and setup errors between 1mm and 5mm were found in the 52.3%. 6.1% patients showed the more than 5mm shift and this error were verified as a difference of setup position and the movement of patient in a treatment room. Conclusion: The setup error analysis using OBI in this study verified that the conventional setup process in accordance with the laser and field light was not enough to get rid of the setup error. The KV images acquired using OBI provided good image quality for comparing with simulation images and much lower patients' exposure dose compared with conventional method of using EPID. These advantages of OBI system which were confirmed in this study proved the accuracy and priority of OBI system in the process of IGRT(Image Guided Radiation Therapy).

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A Study on Digital Culture Phenomenon Shown in the Modernly Fashion Design (현대 패션디자인에 나타난 디지털문화현상)

  • Kim, Jee-Hee
    • Fashion & Textile Research Journal
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    • v.7 no.2
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    • pp.143-152
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    • 2005
  • A concept of 'digital' is changing a living pattern of moderners, with having influence on the whole life of modern society. The purpose of this study is arrange the frame of conformity to the 'fashion as culture' by considering the social and cultural phenomena being shown in relation to digital, which is a concept being watched most for the 21st century and by trying to analyze a tendency of digital culture being shown in the modernly fashion design based on this. The digital culture, which is a concept of generalizing the phenomena of interactional changes in the sub-structure being derived by digital technology, is being shown as a tendency of fusionization and globalization, and due to this, the culture of digital nomads is being formed. On the other hand, a tendency of amenity caused by the reaction against the coldly digital technology, is forming one axis of digital culture. As the culture, which experiences the process of a change by digital technology, is reflected even on the fashion, the fusion of technology and the human body, brought about the appearance and the development of the artificial body, by allowing the wearable computer to be introduced to fashion and by being connected directly to the body. This means the expansion of range for fashion. The destruction of a border between space and space, is making an opportunity of forming another ego inside the cyber space, with bringing about the mixed loading between the cyber space and the real space. As the border between the cyber space and the real space is being collapsed, the space of newly self-realization is being created. The collapse of gender is being shown as the pursuit of gender, which is a nomadic concept of not giving priority to anywhere of male gender and female gender. The tendency of sensitive design introduced the sports look as the largely fashion trend. Fascinated with Zen thoughts is leading to a response to the swiftly and coldly social conditions caused by machine. The digital culture by digital technology and the fashion tendency being shown by its influence, meet the needs of self-realization and self-expansion for a human being, and satisfy the needs for the expression of self-identity for a human being, and enable the search for introspection about inner existence inside the self.

Anatomical Studies on the Skeleton of Thoracic Limb of Korean Native Goat (한국재래산양의 전지골격에 관한 해부학적 연구)

  • Kim, Jin-sang;Lee, Heung-shik S.;Lee, In-se
    • Korean Journal of Veterinary Research
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    • v.27 no.2
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    • pp.167-183
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    • 1987
  • The anatomical structure of the Skeleton of thoracic limb of thirty-one adult Korean native goats(body weight: 14~17kg) was observed after skeletal preparation, and the osteometry was performed in each bone. The results were as follows; 1. The thoracic limb of the Korean native goat was composed of scapula, humerus, radius, ulna, carpal bones, metacarpal bones, phalanges and sesamoid bones. 2. The scapula was flat and triangular in shape. There were no distinct tuber of spine and acromion in the spine. The subscapular fossa was deep and triangular in shape and the vertebral border was sigmoid form. The coracoid bone was formed as the coracoid process at the medial aspect of the supraglenoid tubercle but the clavicle wa.s not observed. The left and right scapular indexes were 57.92 and 58.31 and the glenoid cavity indexes were 89.23 and 86.82, respectively. 3. The greater tubercle of the humerus was devided into cranial and caudal parts. The third tubercle was observed and the face for the infraspinatus muscle was rectangular form. The left and right humerus indexes were 32.44 and 32.63, the head indexes were 94.13, 96.62 and the trochlear-epidondyle indexes were 67.32 and 65.81, respectively. 4. The radius and ulna were fused entirely except at the broad proximal and narrow distal interosseous spaces. The ulna was longer than the radius, and its reduced body and distal end were fused at the caudomedial surface of the radius. 5. The carpal bones were six in number. There were radial, intermediate, ulnar, accessory, second-third and fourth carpal hones in carpal bones. 6. The metacarpal bone was composed of a large metacarpal bone resulted from the fusion of the third and fourth metacarpal bones, and there was a metacarpal tubercle at the dorsolateral part of the proximal end. There were no vestiges of the second and fifth metacarpal bones. 7. The digits were composed of third and fourth digits and each digit was composed of the proximal, middle and distal phalanges. 8. The sesamoid bones were six in number. There were two at the fetlock joint and one at the coffine joint palmarly in each digit. 9. The ratios of the lengths among the scapula, humerus, antebrachium and metacarpal bone were 1.42 : 1.47 : 1.77 : 1.00 in the left and 1.42 : 1.45 : 1.77 : 1.00 in the right, respectively.

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Establishment of a Stable Cell Line Expressing Human BMP2/7-PTD for Efficient Osteogenic Induction (효과적인 뼈 세포분화 유도를 위한 유전자 재조합 PTD 융합 인간 뼈 형성촉진인자2/7(BMP2/7-PTD)를 발현하는 세포주 개발)

  • Park, Seung-Won;Kang, Seok-Woo;Goo, Tae-Won;Kim, Seong-Ryul;Paik, Soon-Young
    • Journal of Life Science
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    • v.22 no.4
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    • pp.456-465
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    • 2012
  • Heterodimeric recombinant human bone morphogenetic proteins (rhBMPs) are powerful tools for bone tissue engineering. However, BMPs have several important limitations in their application to bone regeneration. BMPs have a short half-life and must be used in high concentrations, which may be cost-inefficient. To overcome these problems, we established a stable cell line that expressed the fusion protein comprised of recombinant human BMP2/7 heterodimer protein and PTD (rhBMP2/7-PTD). This stable cell line enabled high process yields by continuously expressing rhBMP2/7-PTD products at high levels throughout cultivation. This synthesized BMP7 was fused to a BMP2 protein with four glycine residues (to allow free bond rotation of the domains) and PTD. To demonstrate that the rhBMP2/7-PTD protein that was secreted from an rhBMP2/7-PTD-expressing stable cell line exhibited biological activity consistent with its role as an osteogenic differentiation induction growth factor, we evaluated BMP-induced ALP activity. Our results suggest that this cell line may be a powerful and efficient tool for applications such as bone tissue regeneration.

A Critical analysis of NCS-based Curriculum (국가직무능력표준(NCS) 기반 교육과정에 대한 비판적 고찰)

  • Ko, Kyoung-Im
    • Journal of Digital Convergence
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    • v.13 no.8
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    • pp.69-82
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    • 2015
  • This article critically examines the backgrounds and meanings of National Competency Standards (NCS) that is employed by Korean colleges for managing performance-based, competency-based curriculum. Findings are as follows: 1) the NCS-based curriculum was primarily adopted to enhance students' business competency for their successes in a competitive job market. 2) NCS is questioned its efficacy to resolve Korean employment issues in a serious economic structure in which a growing number of jobless youth and aged employees and education discrimination are involved. 3) NCS, with its emphasis on the Tyler Rationale and principles of scientific curriculum development, has many criticisms due to its technical approaches to educational processes and needs to be replaced with an alternative paradigm. 4) This article suggests that administrators, policy makers, and educators seek ways to resolve NCS issues considering contextual features of Korean job market and rethinking NCS ideology in the education process. A need for curriculum reconceptualisation is discussed.

Anti-inflammation Effect of Gynura Procumbens extract (명월초 추출물의 항염 효과에 대한 연구)

  • Jeon, Hyeong-Ju;Kwon, Hye-Jin
    • Journal of Digital Convergence
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    • v.14 no.10
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    • pp.515-520
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    • 2016
  • This study conducted the research about the positive effects of Gynura Procumbens on preventing dermatitis and infectious diseases the contemporary people have. To do it, this study measured the concentration of NF-kB, the extract of Gynura Procumbens. In this regard, cytosol, the extract of Gynura Procumbens, and nucleus went through the separation process. The concentration of major NF-kB among various factors to control inflammation was measured with the use of HDF cell. Regarding RAW264.7 cell, the amounts of NO to play an important role in reacting the skin immune system as the media of neural transmission were analyzed. The outcome about the extracts of Gynura Procumbens injected show that it can be expected that as the NF-kB protein and mRNA band were reduced, Gynura Procumbens would have anti-inflammatory effects that could contribute to preventing dermatitis and diseases. In addition, the extracts of Gynura Procumbens have significantly reduced NO with their concentration increasing. In other words, Gynura Procumbens are considered to regulate dependently the production of NO in the concentration of extracts. Thus there is an expectation that the more intensive research would be conducted to heal dermatitis. And it is deemed that Gynura Procumbens would be used as materials for cosmetics as well as foods that the contemporary people can widely consume if a more careful research about anti-inflammatory effects would be sustained on a human bodies' clinical level.