• Title/Summary/Keyword: Vector Potential

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Transduced HSP27 protein protects neuronal cell death by enhancing FALS-associated SOD1 mutant activity

  • An, Jae-Jin;Lee, Yeom-Pyo;Kim, Dae-Won;Sohn, Eun-Joung;Jeong, Hoon-Jae;Kang, Hye-Won;Shin, Min-Jae;Kim, Mi-Jin;Ahn, Eun-Hee;Jang, Sang-Ho;Kang, Jung-Hoon;Kang, Tae-Cheon;Won, Moo-Ho;Kwon, Oh-Shin;Cho, Sung-Woo;Lee, Kil-Soo;Park, Jin-Seu;Eum, Won-Sik;Choi, Soo-Young
    • BMB Reports
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    • v.42 no.3
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    • pp.136-141
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    • 2009
  • Familial Amyotrophic lateral sclerosis (FALS) is a progressive neurodegenetative disorder induced by mutations of the SOD1 gene. Heat shock protein 27 (HSP27) is well-defined as a stress-inducible protein, however the its role in ALS protection has not yet been established. To investigate the role HSP27 may have in SOD1 mutant-mediated apoptosis, human SOD1 or HSP27 genes were fused with a PEP-1 peptide in a bacterial expression vector to produce a genetic in-frame fusion protein, which was then transduced into cells. We found the purified PEP-1-HSP27 fusion proteins can be transduced efficiently into neuronal cells and protect against cell death by enhancing mutant SOD1 activity. Moreover, transduced PEP-1-HSP27 efficiently prevents protein aggregation produced by oxidative stress. These results suggest that transduced HSP27 fusion protein may be explored as a potential therapeutic agent for FALS patients.

Feasibility as a Laundry Detergent Additive of an Alkaline Protease from Bacillus clausii C5 Transformed by Chromosomal Integration (Chromosomal Integration에 의해 제조한 Bacillus clausii C5 유래의 alkaline protease의 세제 첨가제 응용성)

  • Joo, Han-Seung;Choi, Jang Won
    • KSBB Journal
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    • v.27 no.6
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    • pp.352-360
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    • 2012
  • Bacillus clausii I-52 which produced SDS- and $H_2O_2$-tolerant extracellular alkaline protease (BCAP) was isolated from heavily polluted tidal mud flat of West Sea in Incheon, Korea and stable strain (transformant C5) of B. clausii I-52 harboring another copy of BCAP gene in the chromosome was developed using the chromosome integration vector, pHPS9-fuBCAP. When investigated the production of BCAP using B. clausii transformant C5 through pilot-scale submerged fermentation (500 L) at $37^{\circ}C$ for 30 h with an aeration rate of 1 vvm and agitation rate of 250 rpm, protease yield of approximately 105,700 U/mL was achieved using an optimized medium (soybean meal 2%, wheat flour 1%, sodium citrate 0.5%, $K_2HPO_4$ 0.4%, $Na_2HPO_4$ 0.1%, NaCl 0.4%, $MgSO_4{\cdot}7H_2O$ 0.01%, $FeSO_4{\cdot}7H_2O$ 0.05%, liquid maltose 2.5%, $Na_2CO_3$ 0.6%). The enzyme stability of BCAP was increased by addition of polyols (10%, v/v) and also, the stabilities of BCAP towards not only the thermal-induced inactivation at $50^{\circ}C$ but also the SDS and $H_2O_2$-induced inactivation at $50^{\circ}C$ were enhanced. Among the polyols examined, the best result was obtained with propylene glycol (10%, v/v). The BCAP supplemented with propylene glycol exhibited extreme stability against not only the detergent components such as ${\alpha}$-orephin sulfonate (AOS) and zeolite but also the commercial detergent preparations. The granulized enzyme of BCAP was prepared with approximately 1,310,000 U/g of granule. Wash performance analysis using EMPA test fabrics revealed that BCAP granule exhibited high efficiency for removal of protein stains in the presence of anionic surfactants as well as bleaching agents. When compared to Savinase 6T$^{(R)}$ and Everlase 6T$^{(R)}$ manufactured by Novozymes, BCAP under this study probably showed similar or higher efficiency for the removal of protein stains. These results suggest that the alkaline protease produced from B. clausii transformant C5 showing high stability against detergents and high wash performance has significant potential and a promising candidate for use as a detergent additive.

Construction of BLAST Server for Mollusks (연체동물 전용 서열 블라스트 서버구축)

  • Lee, Yong-Seok;Jo, Yong-Hun;Kim, Dae-Soo;Kim, Dae-Won;Kim, Min-Young;Choi, Sang-Haeng;Yon, Jei-Oh;Byun, In-Sun;Kang, Bo-Ra;Jeong, Kye-Heon;Park, Hong-Seog
    • The Korean Journal of Malacology
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    • v.20 no.2
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    • pp.165-169
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    • 2004
  • The BLAST server for the mollusk was constructed on the basis of the Intel Server Platform SC-5250 dual Xeon 2.8 GHz cpu and Linux operating system. After establishing the operating system, we installed NCBI (National Center for Biotechnology Information) WebBLAST package after web server configuration for cgi (common gate interface) (http://chimp.kribb.re.kr/mollusks). To build up the stand alone blast, we conducted as follows: First, we downloaded the genome information (mitochondria genome information), DNA sequences, amino acid sequences related with mollusk available at NCBI. Second, it was translated into the multifasta format that was stored as database by using the formatdb program provided by NCBI. Finally, the cgi was used for the Stand Alone Blast server. In addition, we have added the vector, Escherichia coli, and repeat sequences into the server to confirm a potential contamination. Finally, primer3 program is also installed for the users to design the primer. The stand alone BLAST gave us several advantages: (1) we can get only the data that agree with the nucleotide sequence directly related with the mollusks when we are searching BLAST; (2) it will be very convenient to confirm contamination when we made the cDNA or genomic library from mollusks; (3) Compared to the current NSBI, we can quickly get the BLAST results on the mollusks sequence information.

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The Change of c-jun Promoter Activity in TPA-Induced U937 Cells Infected with Human Cytomegalovirus (HCMV) (TPA로 분화된 U937 세포에서 사람 세포거대바이러스에 의한 c-jun Promoter 활성도의 변화)

  • Park, Chung-Gyu;Kim, Dae-Joong;Kim, Jin-Hee;Han, Tae-Hee;Hwan, Eung-Soo;Choi, Myong-Sik;Kook, Yoon-Hoh;Choi, Sung-Bae;Cha, Chang-Yong
    • The Journal of Korean Society of Virology
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    • v.29 no.2
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    • pp.129-136
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    • 1999
  • Transient transfection assay has been done to evaluate whether the c-jun activation would be prerequisite to the induction of permissiveness against human cytomegalovirus using in vitro cell model in which U937 has been induced to express CD11b and CD14 to become potential monocyte/macrophage cells by TPA treatment. U937 cells were treated with $10\;{\mu}M$, $50\;{\mu}M$ or $100\;{\mu}M$ of TPA. The cell morphology change was observed and the expression of the CD11b and CD14 was confirmed by FACS. Differentiated cells were transfected with pJLuc reporter vector which contained the wild type murine c-jun promoter spanning the SP1, CTF, ATF/CREB and MEF-2 binding sites upstream of the firefly luciferase gene. After 48 hrs of transfection, the cells were infected with HCMV Towne strain and the luciferase activity was assessed at 1 hand 4 h pi. The transfection assay showed no activation of the c-jun promoter at 1 h pi, instead, it showed 2 times increase of the its activity at 4 h pi. There was no difference of the c-jun promoter activation between TPA treated and untreated U937 cells, implying that c-jun activation might not be prerequisite for allowing cells to be premissive to HCMV, although HCMV infection itself could activate c-jun promoter.

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Molecular Identification and Sequence Analysis of Coat Protein Gene of Ornithogalum mosaic virus Isolated from Iris Plant

  • Yoon, Hye-In;Ryu, Ki-Hyun
    • The Plant Pathology Journal
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    • v.18 no.5
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    • pp.251-258
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    • 2002
  • A potyvirus was isolated from cultivated Iris plants showing leaf streak mosaic symptom. Reverse transcription and polymerase chain reaction (RT-PCR) product of 1 kb long which encoded partial nuclear inclusion B and N-terminal region of viral coat protein (CP) genes for potyviruses was successfully amplified with a set of potyvirus-specific degenerate primers with viral RNA samples from the infected leaves: The RT-PCR product was cloned into the plasmid vector and its nucleotide sequences were determined. The nucleotide sequence of a CDNA clone revealed that the virus was an isolate of Ornithogalum moseic virus (OrMV) based on BLAST search analysis and was denoted as OrMV Korean isolate (OrMV-Ky). To further characterize the CP gene of the virus, a pair of OrMV-specific primers was designed and used for amplification of the entire CP gene of OrMV-Kr, The virus was easily and reliably detected from virus-infected Iris leaves by using the RT-PCR with the set of virus-specific primers. The RT-PCR product of the CP gene of the virus was cloned and its sequences were determined from selected recombinant CDNA clones. Sequence analysis revealed that the CP of OrMV-Kr consisted of 762 nucleotides, which encoded 253 amino acid residues. The CP of OrMV-Ky has 94.1-98.0% amino acid sequence identities (20 amino acid alterations) with that of other three isolates of OrMV, Two NT rich potential N-glycosylation motif sequences, NCTS and NWTM, and a DAC triple box responsible for aphid transmission were conserved in CPs of all the strains of OrMV. The virus has 58.5-86.2% amino acid sequence identities with that of other 16 potyviruses, indicating OrMV to be a distinct species of the genus. OrMV-Ky was the most related with Pterostylia virus Yin the phylogenetic tree analysis of CP at the amino acid level. This is the first report on the occurrence of OrMV in Iris plants in Korea. Data in this study indicate that OrMV is found in cultivated Iris plants, and may have mixed infection of OrMV and Iris severe mosaic virus in Korea.

Program Design and Implementation for Efficient Application of Heterogeneous Spatial Data Using GMLJP2 Image Compression Technique (GMLJP2 영상압축 기술을 이용한 다양한 공간자료의 효율적인 활용을 위한 프로그램 설계 및 구현)

  • Kim, Yoon-Hyung;Yom, Jae-Hong;Lee, Dong-Cheon
    • Journal of the Korean Society of Surveying, Geodesy, Photogrammetry and Cartography
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    • v.24 no.5
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    • pp.379-387
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    • 2006
  • The real world is spatially modelled conceptually either as discrete objects or earth surface. The generated data models are then usually represented as vector and raster respectively. Although there are limited cases where only one data model is sufficient to solve the spatial problem at hand, it is now generally accepted that GIS should be able to handle various types of data model. Recent advances in spatial technology introduced even more variety of heterogeneous data models and the need is ever growing to handle and manage efficiently these large variety of spatial data. The OGC (Open GIS Consortium), an international organization pursuing standardization in the geospatial industry. recently introduced the GMLJP2 (Geographic Mark-Up Language JP2) format which enables store and handle heterogeneous spatial data. The GMLJP2 format, which is based on the JP2 format which is an abbreviation for JPEG2000 wavelet image compression format, takes advantage of the versatility of the GML capabilities to add extra data on top of the compressed image. This study takes a close look into the GMLJP2 format to analyse and exploit its potential to handle and mange hetergeneous spatial data. Aerial image, digital map and LIDAR data were successfully transformed end archived into a single GMLJP2 file. A simple viewing program was made to view the heterogeneous spatial data from this single file.

Construction of a Full-length cDNA Library from Korean Stewartia (Stewartia koreana Nakai) and Characterization of EST Dataset (노각나무(Stewartia koreana Nakai)의 cDNA library 제작 및 EST 분석)

  • Im, Su-Bin;Kim, Joon-Ki;Choi, Young-In;Choi, Sun-Hee;Kwon, Hye-Jin;Song, Ho-Kyung;Lim, Yong-Pyo
    • Horticultural Science & Technology
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    • v.29 no.2
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    • pp.116-122
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    • 2011
  • In this study, we report the generation and analysis of 1,392 expressed sequence tags (ESTs) from Korean Stewartia (Stewartia koreana Nakai). A cDNA library was generated from the young leaf tissue and a total of 1,392 cDNA were partially sequenced. EST and unigene sequence quality were determined by computational filtering, manual review, and BLAST analyses. Finally, 1,301 ESTs were acquired after the removal of the vector sequence and filtering over a minimum length 100 nucleotides. A total of 893 unigene, consisting of 150 contigs and 743 singletons, was identified after assembling. Also, we identified 95 new microsatellite-containing sequences from the unigenes and classified the structure according to their repeat unit. According to homology search with BLASTX against the NCBI database, 65% of ESTs were homologous with known function and 11.6% of ESTs were matched with putative or unknown function. The remaining 23.2% of ESTs showed no significant similarity to any protein sequences found in the public database. Annotation based searches against multiple databases including wine grape and populus sequences helped to identify putative functions of ESTs and unigenes. Gene ontology (GO) classification showed that the most abundant GO terms were transport, nucleotide binding, plastid, in terms biological process, molecular function and cellular component, respectively. The sequence data will be used to characterize potential roles of new genes in Stewartia and provided for the useful tools as a genetic resource.

Psychology analyzing system using spectrum component density ratio of EEG based on BCI-TAT (EEG 대역별 스펙트럼 활성 비를 활용한 BCI-TAT 기반 심리 분석 시스템)

  • Shin, Jeon-Hoon;Jin, Sang-Hyeon
    • Journal of the Institute of Convergence Signal Processing
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    • v.11 no.2
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    • pp.112-124
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    • 2010
  • Studies that can find resolutions to problems of subjective psychiatric analysis must be performed and indeed they are in the process. However there still lies many problems in researches of mentality examination, which should be the foundation of finding potential resolutions. One of the biggest problems in the conventional system is that there are many different opinions by psychiatrists depending on their educations and experiences. There is no objective standard on the subjects and there is no effective psychiatric analysis method. Also, the characteristic of such examinations is that it cannot be applied to disabilities, foreigners and infants alyce the examination is ch examinconversation. The objective of this atudy is to standardize TAT(Thematic Apperception Test)analysiBallken index method so that subjective data from the examination can be excluded and the examination thus maklysithe examination objectified. Furthermore, objective result and patients' brain wave pattern is read with BCI(Brain Computer Interface) ch exaTherenvironment to Alsare it to brain wave characteristics vectors to reate brain-wave characteristics vector DB. Therefore, such DB can be utilize durlysithe examination and diagnosis to reate objective examination method and standardized diagnosis system. Thus, mentality examination can be performed only with brain-wave scans with BCI based TAT system.

Expression Profiles of Streptomyces Doxorubicin Biosynthetic Gene Cluster Using DNA Microarray System (DNA Microarray 시스템을 이용한 방선균 독소루비신 생합성 유전자군의 발현패턴 분석)

  • Kang Seung-Hoon;Kim Myung-Gun;Park Hyun-Joo;Kim Eung-Soo
    • KSBB Journal
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    • v.20 no.3
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    • pp.220-227
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    • 2005
  • Doxorubicin is an anthracycline-family polyketide compound with a very potent anti-cancer activity, typically produced by Streptomyces peucetius. To understand the potential target biosynthetic genes critical for the doxorubicin everproduction, a doxorubicin-specific DNA microarray chip was fabricated and applied to reveal the growth-phase-dependent expression profiles of biosynthetic genes from two doxorubicin-overproducing strains along with the wild-type strain. Two doxorubicin-overproducing 5. peucetius strains were generated via over-expression of a dnrl (a doxorubicin-specific positive regulatory gene) and a doxA (a gene involved in the conversion from daunorubicin to doxorubicin) using a streptomycetes high expression vector containing a strong ermE promoter. Each doxorubicin-overproducing strain was quantitatively compared with the wild-type doxorubicin producer based on the growth-phase-dependent doxorubicin productivity as well as doxorubicin biosynthetic gene expression profiles. The doxorubicin-specific DNA microarray chip data revealed the early-and-steady expressions of the doxorubicin-specific regulatory gene (dnrl), the doxorubicin resistance genes (drrA, drrB, drrC), and the doxorubicin deoxysugar biosynthetic gene (dnmL) are critical for the doxorubicin overproduction in S. peucetius. These results provide that the relationship between the growth-phase-dependent doxorubicin productivity and the doxorubicin biosynthetic gene expression profiles should lead us a rational design of molecular genetic strain improvement strategy.

MAXILLARY ADVANCEMENT USING RIGID EXTERNAL DISTRACTION(RED) IN CLEFT LIP AND PALATE PATIENT : CASE REPORT (견고 구외 골신장술을 이용한 구순구개열 환아의 치험례)

  • Yu, Nan-Young;Kim, Sung-Min;Lee, Ju-Hyun;Seo, Hyun-Woo;Park, Ho-Won
    • Journal of the korean academy of Pediatric Dentistry
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    • v.32 no.4
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    • pp.709-716
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    • 2005
  • Patients with cleft tip and palate present severe maxillary hypoplasia due to scar of lip and palate, often accompanied by compromised mastication, speech abnormalities. Sometimes maxillary hypoplasia persist even though active orthodontic treatment was done. In theses cases, patients born with cleft lip and palate will be potential candidates for maxillary advancement with bone grafting after growth to correct the functional deformities and improve aesthetic facial proportions. But, maxillary advancement using standard surgical approaches has several limitations : increased relapse tendency after maxillary advancement, necessity of additional bone graft and mandibular setback surgery. Distraction osteogenesis is current treatment modality to overcome these limitations, thus has become popular for treatment of maxillary hypoplasia associated cleft lip and palate, craniosyntosis. Especially, rigid external distraction, contrary to internal device, has advantages : better vector control of osteotomized segment, effective traction of the bony segments, the ease of the application and removal the distraction device. This study showed that relatively successful result could be generated by using rigid external distraction osteogenesis(RED) in the case of cleft lip and palate with severe maxillary hypoplasia, 6 years 7 months old.

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