• BMB Reports
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• v.39 no.1
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• pp.55-60
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• 2006

We describe a phage display strategy, based on the differential resistance of proteins to denaturant-induced unfolding, that can be used to select protein variants with improved conformational stability. To test the efficiency of this strategy, wild-type and two stable variants of ${\alpha}_1$-antitrypsin (${\alpha}_1AT$) were fused to the gene III protein of M13 phage. These phages were incubated in unfolding solution containing denaturant (urea or guanidinium chloride), and then subjected to an unfavorable refolding procedure (dialysis at $37^{\circ}C$). Once the ${\alpha}_1AT$ moiety of the fusion protein had unfolded in the unfolding solution, in which the denaturant concentration was higher than the unfolding transition midpoint ($C_m$) of the ${\alpha}_1AT$ variant, around 20% of the phage retained binding affinity to anti-${\alpha}_1AT$ antibody due to a low refolding efficiency. Moreover, this affinity reduced to less than 5% when 10 mg/mL skimmed milk (a misfolding-promoting additive) was included during the unfolding/refolding procedure. In contrast, most binding affinity (>95%) remained if the ${\alpha}_1AT$ variant was stable enough to resist unfolding. Because this selection procedure does not affect the infectivity of M13, the method is expected to be generally applicable to the high-throughput screening of stable protein variants, when activity-based screening is not possible.

• Journal of Microbiology and Biotechnology
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• v.26 no.12
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• pp.2184-2191
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• 2016

The time-consuming and high-cost preparation of soluble peptide-major histocompatibility complexes (pMHC) currently limits their wide uses in monitoring antigen-specific T cells. The single-chain trimer (SCT) of peptide-${\beta}2m$-MHC class I heavy chain was developed as an alternative strategy, but its gene fusion is hindered in many cases owing to the incompatibility between the multiple restriction enzymes and the restriction endonuclease sites of plasmid vectors. In this study, overlap extension PCR and one-step cloning were adopted to overcome this restriction. The SCT gene of the $OVA_{257-264}$ peptide-$(GS_4)_3-{\beta}2m-(GS_4)_4-H-2K^b$ heavy chain was constructed and inserted into plasmid pET28a by overlap extension PCR and one-step cloning, without the requirement of restriction enzymes. The SCT protein was expressed in Escherichia coli, and then purified and refolded. The resulting $H-2K^b/OVA_{257-264}$ complex showed the correct structural conformation and capability to bind with $OVA_{257-264}$-specific T-cell receptor. The overlap extension PCR and one-step cloning ensure the construction of single-chain MHC class I molecules associated with random epitopes, and will facilitate the preparation of soluble pMHC multimers.

• Journal of Fashion Business
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• v.22 no.5
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• pp.64-82
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• 2018

As sustainability of fashion brands is becoming more important in the prolonged depression, the fashion market is also seeking ways to create new value through convergence with various fields. In the fashion market, convergence marketing is engaged with diverse technologies combined with advanced technologies and strategies. This study established the conceptual characteristics of convergence and divergence from a market perspective. Convergence can be summarized as Borderless, Compounding, Diversification, Recombination and Affiliation. The conceptual characteristics of the divergence can be summarized as de-fusion, individualization, differentiation, specialization, and segmentation. The characteristics of convergence revealed through the case of fashion market include the expandability of the new category, the expandability of the consumer, the brand experience, and the expandability of time and space. Divergence is the differentiation of brand value, target customer, and product nature. Through this study, we could understand the differentiation of the fashion market and the direction of the marketing strategy by looking at the characteristic of the convergence marketing - oriented research that has the contrary conceptual characteristics from the extended viewpoint. Based on this, it is possible to see the changes and patterns of brand marketing strategies and techniques in recent fashion market. In the future, it is expected that the systematic study of the marketing strategy of the domestic fashion brand based on the convergence and change characteristics of the fashion market proposed in this study will continue.

• Journal of the Korea Society of Computer and Information
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• v.25 no.12
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• pp.271-278
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• 2020

This study looked at the World Martial Arts Mastership's 'non-face promotional media strategies and methods' after the Corona-19 fandom, and found out if they could be applied to other sports events. As a method of research, the promotional media strategy of martial arts mastership was divided into types and in-depth interviews with media agents were conducted. According to the study, hosting an online martial arts competition, building an untact convention platform, and a base education facility can be a successful contest with budget support. Martial arts branding, popularization and industrialization can be a new attempt if ideas and execution personnel are sufficient. If IT technology and Chungbuk martial arts infrastructure are combined, they will be able to pioneer a new model called 'non-face-to-face-to K-sports'.

• 한국디지털정책학회:학술대회논문집
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• 2006.12a
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• pp.379-386
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• 2006

The purpose of this paper is to analyze the implementation of the GIS infrastructure systems for the u-City, GIS base u-City represents spatial information derived fields, such as geographical distribution of the urban boundaries, physical configuration of the urban locations and cultural characteristics of the urban history. These three aspects relate to urban infrastructure systems implementation, urban monitoring center implementation, and spatial database implementation. In terms of the GIS based u-Ci쇼 urban infrastructure implementation systems, the u-City depends on IT contents and spatial features. IT contents are strongly related to IT839 strategy due to the national agenda is "u-Korea". GIS should contribute to u-City construction through the spatial analyses methods. For these methods various GIS functions will guide to u-City's distribution, location, and characteristics of urbanization. The infrastructure consists of road and road facilities, underground facilities, related agencies facilities, dispatch systems, environmental systems, and urban planning. These six units of the urban infrastructures have spatial databases that consist of spatial configuration, such as dots, lines, and polygons in order to draw the spatial distribution of the u-City GIS based u-City urban infrastructure implementation systems should deal with It convergence to generate fusion affects.

• Journal of the Korean Institute of Telematics and Electronics B
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• v.29B no.11
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• pp.120-129
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• 1992

In this paper we propose a fuzzy-neural network(FNN) which includes both advantages of the fuzzy logic and the neural network. The basic idea of the FNN is to realize the fuzzy rule-base and the process of reasoning by neural network and to make the corresponding parameters be expressed by the connection weights of neural network. After constructing the FNN, a novel controller consisting of a conventional P-controller and a FNN is explained. In this control scheme, the rule-base of a FNN are automatically generated by error back-propagation algorithm. Also the parallel connection of the P-controller and the FNN can guarantee the stability of a plant at initial stage before the rules are completely created. Finally the effectiveness of the proposed strategy will be verified by computer simulations using a 2 degree of freedom robot manipulator.

• Journal of Microbiology and Biotechnology
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• v.19 no.12
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• pp.1620-1627
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• 2009

The design and expression of an antihypertensive peptide multimer (AHPM), a common precursor of 11 kinds of antihypertensive peptides (AHPs) tandemly linked up according to the restriction sites of gastrointestinal proteases, were explored. The DNA fragment encoding the AHPM was chemically synthesized and cloned into expression vector pGEX-3X. After an optimum induction with IPTG, the recombinant AHPM fused with glutathione S-transferase (GST-AHPM) was expressed mostly as inclusion body in Escherichia coli BL21 and reached the maximal production, 35% of total intracellular protein. The inclusion body was washed, dissolved, and purified by cation-exchange chromatography under denaturing conditions, followed by refolding together with size-exclusion chromatography and gradual dialysis. The resulting yield of the soluble GSTAHPM (34 kDa) with a purity of 95% reached 399 mg/l culture. The release of high active fragments from the AHPM was confirmed by the simulated gastrointestinal digestion. The results suggest that the design strategy and production method of the AHPM will be useful to obtain a large quantity of recombinant AHPs at a low cost.

• Journal of Microbiology and Biotechnology
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• v.26 no.7
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• pp.1267-1277
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• 2016

Currently, enzymatic synthesis of lactulose, a synthetic prebiotic disaccharide, is commonly performed with glycosyl hydrolases. In this work, a new type of lactulose-producing biocatalyst was developed by displaying β-galactosidase from Bacillus stearothermophilus IAM11001 (Bs-β-Gal) on the surface of Bacillus subtilis 168 spores. Localization of β-Gal on the spore surface as a fusion to CotX was verified by western blot analysis, immunofluorescence microscopy, and flow cytometry. The optimum pH and temperature for the resulting spore-displayed β-Gal was 6.0 and 75℃, respectively. Under optimal conditions, it showed maximum activity of 0.42 U/mg spores (dry weight). Moreover, the spore-displayed CotX-β-Gal was employed as a whole cell biocatalyst to produce lactulose, yielding 8.8 g/l from 200 g/l lactose and 100 g/l fructose. Reusability tests showed that the spore-displayed CotX-β-Gal retained around 30.3% of its initial activity after eight successive conversion cycles. These results suggest that the CotX-mediated spore-displayed β-Gal may provide a promising strategy for lactulose production.

• International Journal of Advanced Culture Technology
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• v.4 no.4
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• pp.81-86
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• 2016

In this paper, we propose a simple and effective vision-based tracking controller design for autonomous object tracking using multicopter. The multicopter based automatic tracking system usually unstable when the object moved so the tracking process can't define the object position location exactly that means when the object moves, the system can't track object suddenly along to the direction of objects movement. The system will always looking for the object from the first point or its home position. In this paper, PID control used to improve the stability of tracking system, so that the result object tracking became more stable than before, it can be seen from error of tracking. A computer vision and control strategy is applied to detect a diverse set of moving objects on Raspberry Pi based platform and Software defined PID controller design to control Yaw, Throttle, Pitch of the multicopter in real time. Finally based series of experiment results and concluded that the PID control make the tracking system become more stable in real time.

• Tunnel and Underground Space
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• v.23 no.4
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• pp.271-279
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• 2013

This report introduced the types of drilling equipment and their operation mechanisms. The state of the art technologies of the Top-hammer drill equipment were investigated and the technology level of Korean drill industry was compared to that of the advanced country. Based on the investigation, the necessity of fusion research and development in the Korean drilling technology and industry was discussed, and the future strategy to catch up with the advanced technology was suggested.