• Journal of Microbiology and Biotechnology
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• 제24권2호
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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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• 제24권6호
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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.

• 한국인쇄학회:학술대회논문집
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• 한국인쇄학회 2001년도 국제학술발표회
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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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• 제25권2호
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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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• 제39권sup2호
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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.

• 한국콘텐츠학회논문지
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• 제11권1호
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• pp.92-101
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• 2011

21세기 기술 혁신으로 3D 디스플레이 기술 및 3D 영상 기술은 새로운 트랜드로 주목 받고 있다. 그러나 3D 입체 영상을 직접 제작하기 위해서는 고가의 제작비용과 전문적인 기술이 필요하다. 3D 디스플레이 장치가 일반화 되면서 사용자 고가의 비용 부담 없이 3D 입체 영상을 구현할 필요성이 증대되고 있다. 본 논문은 싱글 카메라를 이용한 마커 인식 기술을 사용하여 3D 입체 영상을 용이하게 구현 할 방법을 제시하였다. 또한 영상의 해상도를 동적으로 조절가능하게 하였다. 본 논문은 3D 입체 영상 구현 분야에 일반 사용자의 적극적인 참여를 유도하여 3D 입체 영상을 이용한 UCC(User Created Contents) 분야의 활성화에 기여할 것이다.

• 인터넷정보학회논문지
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• 제14권6호
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• pp.9-18
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• 2013

본 논문에서는 모바일 로봇이 주어진 환경에서 물건 객체를 찾기 위해 RFID(Radio Frequency IDentification) 태그의 위치를 추정할 수 있는 지능형 시스템을 설계하고 구현하였다. 개발된 지능형 시스템은 인공 신경망 시스템으로 RSSI(Received Signal Strength Indication)값을 입력으로 하고 절대 위치 좌표 값을 목표 값으로 하는 학습 시스템이다. 위치 추정을 위하여 수동형 RFID를 사용하였으며 능동형으로의 확장도 고려하였다. 실내에서 위치 추정을 위한 환경을 구축하여 사용될 수 있도록 시스템을 설계하였다. 또한 설계된 시스템을 소프트웨어 개발을 하였고, 실험을 통해 테스트베드 상에서 시스템 학습과 관련된 결과를 보여준다. 실제 현장과 유사한 환경에서 학습데이터 생성에서부터 실시간 위치 추정과 관련된 다양한 실험 결과를 보여준다. 실험 결과를 통해 모바일 로봇이 실내에서 원하는 객체를 쉽게 찾을 수 있음을 확인하였다.

• 한국정보통신학회논문지
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• 제15권10호
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• pp.2079-2086
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• 2011

최근 헬스케어 응용 시스템 중의 하나인 자가 진단 시스템 기술의 연구가 활발하게 이루어져 왔다. 이와 함께 본 연구논문은 PC, RFID, 생체신호측정기와 연동된 자가 혈당측정 시스템 및 사용자가 이해하기 쉬운 인터페이스를 구현하는데 있다. 따라서 PC, RFID, 그리고 생체신호측정 센서와 연동되어 손쉽게 자가진단 데이터를 확보할 수 있도록 디바이스와 PC사이의 통신을 위한 손쉬운 인터페이스 구성과 RFID 태그 및 리더기 통신 상태를 분석, 테스트를 수행했다. 즉, 자가 혈당측정 데이터에 대한 통신 프로토콜 수행(프로토콜 정의에 따른 리퀘스트 및 리스폰스 데이터 분석)으로 연월일, 체온 및 혈당수치, 그리고 혈당체크 시점 등 신뢰성 높은 결과가 출력되었음을 나타내었다.

• Molecules and Cells
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• 제39권3호
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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.

• 디지털융복합연구
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• 제17권12호
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• pp.211-217
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• 2019

무선 위치 추적 시스템은 모바일, 로봇 추적 시스템 및 인터넷 위치 기반 서비스(LBS) 애플리케이션과 같은 응용분야에서 사용할 수 있다. GPS 시스템은 가장 잘 알려진 위치 추적 시스템이지만 실내에서의 사용이 용이하지 않다. 실내 위치 추적 시스템에 대한 비용 효율적인 측면에서 무선 주파수 식별(RFID) 위치 추적 방법을 연구하였다. RFID 시스템 대부분은 소모성 배터리를 사용하는 능동형 RFID 태그를 사용지만, 본 논문에서는 패시브 RFID 태그를 사용하는 저렴한 실내 위치 추적 시스템을 개발하였다. 정밀한 추적 알고리즘을 사용하기 위하여 패턴인식에 의한 위치 추적 방법과 회귀곡선을 생성하여 위치를 추적하는 시스템을 연구하였다. 시스템은 잡음으로 인한 오류 수준을 검증하여 테스트하였다. 회귀식에 의해 생성된 3차원 곡선은 미분방정식에 의해서 확률적으로 설명력이 높은 좌표를 추정하였다. 이 제안된 시스템은 모바일 로봇 시스템, AGV 및 휴대전화 LBS에도 적용될 수 있다.