• Journal of Microbiology and Biotechnology
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• v.24 no.2
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• pp.152-159
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• 2014

The regulation of protein tyrosine phosphorylation is mediated by protein tyrosine kinases (PTKs) and protein tyrosine phosphatases (PTPs) and is essential for cellular homeostasis. Co-expression of PTKs with PTPs in Pichia pastoris was used to facilitate the expression of active PTKs by neutralizing their apparent toxicity to cells. In this study, the gene encoding phosphatase PTP1B with or without a blue fluorescent protein or peroxisomal targeting signal 1 was cloned into the expression vector pAG32 to produce four vectors. These vectors were subsequently transformed into P. pastoris GS115. The tyrosine kinases EGFR-2 and $PDGFR{\beta}$ were expressed from vector pPIC3.5K and were fused with a His-tag and green fluorescent protein at the N-terminus. The two plasmids were transformed into P. pastoris with or without PTP1B, resulting in 10 strains. The EGFR-2 and $PDGFR{\beta}$ fusion proteins were purified by $Ni^{2+}$ affinity chromatography. In the recombinant P. pastoris, the PTKs co-expressed with PTP1B exhibited higher kinase catalytic activity than did those expressing the PTKs alone. The highest activities were achieved by targeting the PTKs and PTP1B into peroxisomes. Therefore, the EGFR-2 and $PDGFR{\beta}$ fusion proteins expressed in P. pastoris may be attractive drug screening targets for anticancer therapeutics.

• Journal of Microbiology and Biotechnology
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• v.24 no.6
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• pp.803-811
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• 2014

Concerns over drug-resistant bacteria have stimulated interest in developing alternative methods to control bacterial infections. Endolysin, a phage-encoded enzyme that breaks down bacterial peptidoglycan at the terminal stage of the phage reproduction cycle, is reported to be effective for the control of bacterial pathogenic bacteria. Bioinformatic analysis of the SPN9CC bacteriophage genome revealed a gene that encodes an endolysin with a domain structure similar to those of the endolysins produced by the P1 and P22 coliphages. The SPN9CC endolysin was purified with a C-terminal oligo-histidine tag. The endolysin was relatively stable and active over a broad temperature range (from $24^{\circ}C$ to $65^{\circ}C$). It showed maximal activity at $50^{\circ}C$, and its optimum pH range was from pH 7.5 to 8.5. The SPN9CC endolysin showed antimicrobial activity against only gram-negative bacteria and functioned by cutting the glycosidic bond of peptidoglycan. Interestingly, the SPN9CC endolysin could lyse intact gram-negative bacteria in the absence of EDTA as an outer membrane permeabilizer. The exogenous lytic activity of the SPN9CC endolysin makes it a potential therapeutic agent against gram-negative bacteria.

• Proceedings of the Korean Printing Society Conference
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• 2001.06a
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• pp.63-77
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• 2001

PDF(Portable Document Format) is a file format that Adobe advances postscritp technique and use in managing document information or electric publishing(internet, CD-ROM, DVD). PDF is a devised document type for being able to read and print anywhere, independent of OS, printer type, resolution, and the kind of computer etc. Because this includes a compressing function, it transfers document through a small size of file in internet or intranet. In addition, that is a file format has various advantages-sharing of information and transfering documents in on line or off line environment. In this paper, we developed electronic document system using PDF format. Electronic document system consists of filter, automatic indexing, special searching system and web server. The information used in this paper is database made using Zwon\`s DocuCom. The filter recognizes various kinds of document structure. And according to property of document, it produces ASCII output. In addition to processing various formats of document, the filter can extract keywords in documents of MS WORD, Excel, Powerpoint, PDF, CAD etc. This filter uses the structure of window printer drive and can extract the information for text, page, font type and size from relevant document. The automatic indexing recognizes the formatted tag of document form ASCII text produced by filter and extracts adequate keyword to structure and property of document. PDF electronic document systems proposed in this paper can be used in Internet, PC communication. Users can choose and read electronic documents by two ways. First, users can choose and read relevant books using PDF electronic document homepage. Second, users can use PDF integrated-search system. User can search after inputing keyword and choose reference field and type of data. But, now, PDF products of Adobe can\`t support the Korean character. If this problem is resolved, we thick that PDF applications system looks active. Although there is limited function in case of using Zwon DocuCom used in this study, we think that there isn\`t a great deal of difficulty in electronic document and building digital database.

• Journal of Microbiology and Biotechnology
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• v.25 no.2
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• pp.274-279
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• 2015

Receptor activator of nuclear factor-kappa B ligand (RANKL) is a critical factor in osteoclastogenesis. It makes osteoclasts differentiate and multinucleate in bone remodeling. In the present study, RANKL was expressed as a soluble maltose binding protein (MBP)-fusion protein using the Escherichia coli maltose binding domain tag system (pMAL) expression vector system. The host cell E. coli DH5α was cultured and induced by isopropyl β-_D-1-thiogalactopyranoside for rRANKL expression. Cells were disrupted by sonication to collect soluble MBP-fused rRANKL. The MBP-fusion rRANKL was purified with MBP Trap affinity chromatography and treated with Tobacco Etch Virus nuclear inclusion endopeptidase (TEV protease) to remove the MBP fusion protein. Dialysis was then carried out to remove binding maltose from the cleaved rRANKL solution. The cleaved rRANKL was purified with a second MBP Trap affinity chromatography to separate unsevered MBP-fusion rRANKL and cleaved MBP fusion protein. The purified rRANKL was shown to have biological activity by performing in vitro cell tests. In conclusion, biologically active rRANKL was successfully purified by a simple two-step chromatography purification process with one column.

• Journal of Periodontal and Implant Science
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• v.39 no.sup2
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• pp.231-237
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• 2009

Purpose: Cytolethal distending toxin (CDT) is a family of heat-labile cytotoxins produced by several gram-negative mucosa-associated pathogens, including Aggregatibacter actinomycetemcomitans. CDT is well known to be capable of inducing growth arrest, morphological alterations, and eventually death in various cells. CDT belongs to a tripartite $AB_2$ toxin (CdtB: the enzymatic A subunit; CdtA and CdtC: the heterodimeric B subunit). Previous studies proposed that CdtA and CdtC together bind to a cell surface receptor and glycolipids act as a receptor for A. actinomycetemcomitans CDT (AaCDT). In this study, recombinant CdtA and CdtC proteins of AaCDT were co-expressed in a bacterial expression system and tested for their affinity for $GM_1$ ganglioside. Methods: The genes for CdtA and CdtC from A. actinomycetemcomitans Y4 were utilized to construct the expression vectors, pRSET-cdtA and pET28a-cdtC. Both CdtA and CdtC proteins were expressed in Escherichia coli BL21(DE3) and then purified using hexahistidine (His6) tag. The identity of purified protein was confirmed by anti-His6 antibody and monoclonal anti-CdtA antibody. Furthermore, the affinity of recombinant protein to $GM_1$ ganglioside was checked through ELISA. Results: Recombinant CdtA and CdtC proteins were expressed as soluble proteins and reacted to anti-His6 and monoclonal anti-CdtA antibodies. ELISA revealed that purified soluble CdtA-CdtC protein bound to $GM_1$ ganglioside, while CdtA alone did not. Conclusions: Co-expression of CdtA and CdtC proteins enhanced the solubility of the proteins in E. coli, leading to convenient preparation of active CdtA-CdtC, a critical material for the study of AaCDT pathogenesis.

• The Journal of the Korea Contents Association
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• v.11 no.1
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• pp.92-101
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• 2011

However, in order to create a stereoscopic 3D image, independently, it is required to spend expensive manufacturing costs and to have special techniques. As 3D display devices have been generalized, there is an increasing need for implementing a stereoscopic 3D image without a burden of expensive costs. This paper proposes some methods to implement stereoscopic 3D images easily by utilizing a marker tracking technology using a single camera. In addition, the study made it possible for the resolution of an image to be adjustable dynamically. This paper will be committed to the promotion of the field of UCC (User Created Contents) using stereoscopic 3D images by attracting the active participation of general users in the field of the implementation of stereoscopic 3D images.

• Journal of Internet Computing and Services
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• v.14 no.6
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• pp.9-18
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• 2013

In this paper, we design and implement an intelligent system for finding objects with RFID(Radio Frequency IDentification) tag in which an mobile robot can do. The system we developed is a learning system of artificial neural network that uses RSSI(Received Signal Strength Indicator) value as input and absolute coordination value as target. Although a passive RFID is used for location estimation, we consider an active RFID for expansion of recognition distance. We design the proposed system and construct the environment for indoor location estimation. The designed system is implemented with software and the result related learning is shown at test bed. We show various experiment results with similar environment of real one from earning data generation to real time location estimation. The accuracy of location estimation is verified by simulating the proposed method with allowable error. We prepare local test bed for indoor experiments and build a mobile robot that can find the objects user want.

• Journal of the Korea Institute of Information and Communication Engineering
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• v.15 no.10
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• pp.2079-2086
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• 2011

Recently, technology of self diagnosis system which one of health-care has been research very active. With this, In this paper, we have to implement the self-blood sugar measurement system that is interlock with PC, RFID, and Bio-Signal glucose tester and accessible user interface. Therefore the equipments can performs accessible interface composition for communication between devices and PC, analysis and test of communication status of the RFID tag and reader that it easy to obtain the data of self-diagnosis linkage with PC, RFID, and Bio-Signal glucose tester. In other words, as result of communication protocol experiment with self-blood sugar measurement data, it has been showed to display a high-reliability result through year, month, and day, body temperature, and levels and check point of blood glucose.

• Molecules and Cells
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• v.39 no.3
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• pp.242-249
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• 2016

A balance between production and degradation of reactive oxygen species (ROS) is critical for maintaining cellular homeostasis. Increased levels of ROS during oxidative stress are associated with disease conditions. Antioxidant enzymes, such as extracellular superoxide dismutase (EC-SOD), in the extracellular matrix (ECM) neutralize the toxicity of superoxide. Recent studies have emphasized the importance of EC-SOD in protecting the brain, lungs, and other tissues from oxidative stress. Therefore, EC-SOD would be an excellent therapeutic drug for treatment of diseases caused by oxidative stress. We cloned both the full length (residues 1-240) and truncated (residues 19-240) forms of human EC-SOD (hEC-SOD) into the donor plasmid pFastBacHTb. After transposition, the bacmid was transfected into the Sf9-baculovirus expression system and the expressed hEC-SOD purified using FLAG-tag. Western blot analysis revealed that hEC-SOD is present both as a monomer (33 kDa) and a dimer (66 kDa), as detected by the FLAG antibody. A water-soluble tetrazolium (WST-1) assay showed that both full length and truncated hEC-SOD proteins were enzymatically active. We showed that a potent superoxide dismutase inhibitor, diethyldithiocarbamate (DDC), inhibits hEC-SOD activity.

• Journal of Digital Convergence
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• v.17 no.12
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• pp.211-217
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• 2019

Location positioning systems are available in applications such as mobile, robotic tracking systems and Wireless location-based service (LBS) applications. The GPS system is the most well-known location tracking system, but it is not easy to use indoors. The method of radio frequency identification (RFID) location tracking was studied in terms of cost-effectiveness for indoor location tracking systems. Most RFID systems use active RFID tags using expendable batteries, but in this paper, an inexpensive indoor location tracking system using passive RFID tags has been developed. A pattern matching method and a system for tracing location by generating regression curves were studied to use precision tracking algorithms. The system was tested by verifying the level of error caused by noise. The three-dimensional curves are produced by the regression equation estimated the statistically meaningful coordinates by the differential equation. The proposed system could also be applied to mobile robot systems, AGVs and mobile phone LBSs.