• Korean Journal Plant Pathology
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• v.11 no.1
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• pp.66-72
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• 1995

The coat protein (CP) gene of cucumber mosaic virus-As (CMV-As) strain was engineered for expression in the plant by using the cauliflower mosaic virus 35S transcript regulatory sequences. The CP gene was cloned into an Agrobacterium-derived binary vector. A chimeric gene was constructed by the cDNA of CMV-As CP and plant expression vector pBI121. The clone, pCMAS66, was first introduced into the phagemid vector pSPORT1 for situating sense orientation for translation and making restriction sites in order to re-introduce plant expression vector, pHI121. The resulting subclone pCASCP02 and plant expression vector pBI121 were treated with BamHI-SacI for excising the target gene and removing GUS gene, respectively. After Agrobacterium transformation by freeze-thaw technique, the clone, pCMASCP121-123 which contains sense orientation of the target gene, was selected and confirmed by restriction endonuclease analysis. The CMV-As CP gene was introduced into A. tumefaciens. The results on tobacco plant transformation with the vector system revealed that the system could be successfully introduced and showed high frequency of selection to putative transformations.

• Molecules and Cells
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• v.41 no.10
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• pp.917-922
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• 2018

The CRISPR-Cas system is a well-established RNA-guided DNA editing technique widely used to modify genomic DNA sequences. I used the CRISPR-Cas9 system to change the second and third nucleotides of the triplet $T{\underline{CT}}$ of human TNSFSF9 in HepG2 cells to $T{\underline{AG}}$ to create an amber stop codon. The $T{\underline{CT}}$ triplet is the codon for Ser at the $172^{nd}$ position of TNSFSF9. The two substituted nucleotides, AG, were confirmed by DNA sequencing of the PCR product followed by PCR amplification of the genomic TNFSF9 gene. Interestingly, sequencing of the cDNA of transcripts of the edited TNFSF9 gene revealed that the $T{\underline{AG}}$ had been re-edited to the wild type triplet $T{\underline{CT}}$, and 1 or 2 bases just before the triplet had been deleted. These observations indicate that CRISPR-Cas9-mediated editing of bases in target genomic DNA can be followed by spontaneous re-editing (correcting) of the bases during transcription.

• Molecules and Cells
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• v.43 no.1
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• pp.76-85
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• 2020

MARCH5 is a RING finger E3 ligase involved in mitochondrial integrity, cellular protein homeostasis, and the regulation of mitochondrial fusion and fission. To determine the function of MARCH5 during development, we assessed transcript expression in zebrafish embryos. We found that march5 transcripts were of maternal origin and evenly distributed at the 1-cell stage, except for the mid-blastula transition, with expression predominantly in the developing central nervous system at later stages of embryogenesis. Overexpression of march5 impaired convergent extension movement during gastrulation, resulting in reduced patterning along the dorsoventral axis and alterations in the ventral cell types. Overexpression and knockdown of march5 disrupted the organization of the developing telencephalon and diencephalon. Lastly, we found that the transcription of march5 was tightly regulated by the transcriptional regulators CHOP, C/EBPα, Staf, Znf143a, and Znf76. These results demonstrate the essential role of March5 in the development of zebrafish embryos.

• Journal of Microbiology
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• v.41 no.3
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• pp.239-247
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• 2003

The LRV1-4 capsid protein possesses an endoribonuclease activity that is responsible for the single site-specific cleavage in the 5' untranslated region (UTR) of its own viral RNA genome and the formation of a conserved stem-loop structure (stem-loop IV) in the UTR is essential for the accurate RNA cleavage by the capsid protein. To delineate the nucleotide sequences, which are essential for the correct formation of the stem-loop structure for the accurate RNA cleavage by the viral capsid protein, a wildtype minimal RNA transcript (RNA 5' 249-342) and several synthetic RNA transcripts encoding point-mutations in the stem-loop region were generated in an in vitro transcription system, and used as substrates for the RNA cleavage assay and RNase mapping studies. When the RNA 5' 249-342 transcript was subjected to RNase T1 and A mapping studies, the results showed that the predicted RNA secondary structure in the stem-loop region using FOLD analysis only existed in the presence of Mg$\^$2＋/ ions, suggesting that the metal ion stabilizes the stem-loop structure of the substrate RNA in solution. When point-mutated RNA substrates were used in the RNA cleavage assay and RNase T1 mapping study, the specific nucleotide sequences in the stem-loop region were not required for the accurate RNA cleavage by the viral capsid protein, but the formation of a stem-loop like structure in a region (nucleotides from 267 to 287) stabilized by Mg$\^$2＋/ ions was critical for the accurate RNA cleavage. The RNase T1 mapping and EMSA studies revealed that the Ca$\^$2＋/ and Mn$\^$2＋/ ions, among the reagents tested, could change the mobility of the substrate RNA 5' 249-342 on a gel similarly to that of Mg$\^$2＋/ ions, but only Ca$\^$2＋/ ions identically showed the stabilizing effect of Mg$\^$2＋/ ions on the stem-loop structure, suggesting that binding of the metal ions (Mg$\^$2＋/ or Ca$\^$2＋/) onto the RNA substrate in solution causes change and stabilization of the RNA stem-loop structure, and only the substrate RNA with a rigid stem-loop structure in the essential region can be accurately cleaved by the LRV1-4 viral capsid protein.

• Biomolecules & Therapeutics
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• v.18 no.2
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• pp.171-177
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• 2010

Appetite is inhibited by the anorexigenic neuropeptides POMC (proopiomelanocortin) and CART (cocaine-amphetamine-regulated transcript) in the hypothalamus. The present study was performed to examine the inhibitory effects of baicalin against food intake and the upregulation of POMC/CART. Short-term food intake (48 h) was significantly inhibited by treatment with baicalin (10 mg/kg, p<0.05) in C57BL/6 mice. Immunohistochemical analysis showed that baicalin upregulated POMC and CART levels in the arcuate nucleus of the hypothalamus. These effects were also examined using an in vitro system. pPOMC-Luc or pCART-Luc plasmids were transformed into mouse N29-2 neuronal and human SH-SY5Y cells, and the activities of baicalin were examined in these cells. Baicalin increased POMC and CART promoter-driven luciferase activity in a dose-dependent manner without cytotoxic effects. These results suggest that baicalin downregulates short-term food intake while upregulating POMC and CART expression.

• Proceedings of the Korean Information Science Society Conference
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• 2005.11a
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• pp.898-900
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• 2005

초고속 통신망의 확대 적용으로 인터넷의 빠른 성장과 함께 음성과 비디오 그리고 데이터를 통합하고자 하는 노력이 시도 되고 있다. VoIP(Voice over IP)는 IP를 이용하여 음성과 데이터를 패킷 형태로 통합하여 실시간으로 전송하는 기술이다[1]. 패킷 네트워크에서 VoIP 시그널링 기술을 이용하면 망 자원의 효율적 이용 및 PSTN에 가까운 음질 그리고 인터넷과 연계한 다양한 음성서비스 지원(H.323, SIP, MGCP 등 다양한 신호처리 지원)이 가능하다. 본 논문에서는 VoIP망에서 IP기반 녹취 시스템을 설계 및 구현한다. 녹취 시스템은 고객과 상담원의 통화 내용을 자동으로 녹음하여 보관함으로써 고객의 요구사항을 명확히 파악할 수 있으며 녹취 데이터의 통계 자료 제공으로 효율적인 관리가 지원되고 선택 녹취, 스케줄링 녹취, 상담원의 평가 자료를 제공하여 고객 관리의 질적인 향상을 지원한다. 본 논문의 녹취 시스템은 고객과의 통화 내용을 녹취하여 서버의 녹취 DB에 저장하여 관리하는 녹취 시스템으로 모든 네트워크 환경에서 사용할 수 있으며 CTI와 연동하여 효율적이고 체계적인 녹취 시스템 구국이 가능하다.

• BMB Reports
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• v.49 no.12
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• pp.693-698
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• 2016

In this study, a tissue-specific GAL4/UAS activation tagging system was used for the characterization of genes which could induce lethality when ubiquitously expressed. A dominant mutant exhibiting stunted growth was isolated and named defective root development 1-D (drd1-D). The T-DNA tag was located within the promoter region of AtTX12, which is predicted to encode a truncated nucleotide-binding leucine-rich repeat (NLR) protein, containing a Toll/interleukin-1 receptor (TIR) domain. The transcript levels of AtTX12 and defense-related genes were elevated in drd1-D, and the misexpression of AtTX12 recapitulated the drd1-D phenotypes. In the presence of ENHANCED DISEASE SUSCEPTIBILITY 1 (EDS1), a key transducer of signals triggered by TIR-type NLRs, a low-level of AtTX12 misexpression induced strong defective phenotypes including seedling lethality whereas, in the absence of EDS1, a high-level of AtTX12 misexpression induced weak growth defects like dwarfism, suggesting that AtTX12 might function mainly in an EDS1-dependent and partially in an EDS1-independent manner.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.100.2-101
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• 2003

The global regulator, GacS (global antibiotic and cyanide sensor kinase), was required for the increased resistance to hydrogen peroxide occurring as cultures of the rhizobacterium, P. chlororaphis O6, matured. Specific stationary-phase peroxidase and catalase isozymes were absent in the GacS mutant, whereas a manganese-superoxide dismutase isozyme was expressed earlier and to a great extent than wild type. In the wild type cell, transcript accumulation of rpoS was higher in late logarithmic-phase cells than cells from mid logarithmic- or stationary-phase. Transcripts from rpoS in the GacS mutant were reduced in each of these growth phases compared to the wild type expression. The down stream sequence from rpoS lacked sequences encoding a small RNA, rsmZ, found in other pseudomonads and implicated in control of genes activated by the GacS system. These findings suggest that GacS-mediated regulation of RpoS plays role in control of oxidative stress in P. chlororaphis O6 by as yet an unknown mechanism.

• Journal of Life Science
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• v.11 no.1
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• pp.52-56
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• 2001

The present study intends to characterize the DNA damage-inducible responses in eukaryotic cells. The fission yeast, S. pombe, which displays efficient DNA repair systems, was used in this study as a model system for higher eukaryotes. To study UV-inducible responses in S. pombe, five UV-inducible cDNA clones were isolated from S. pombe by using subtration hybridization method. To investigate the expression of isolated genes, the cellular levels of the transcripts of these genes were determined by Northern blot analysis after UV-irradiation. The transcripts of isolated gene (UV130) increased rapidly and reached maximum accumulation after UV-irradiation. Compared to the message levels of control, the levels of maximal increase were approximately 5 fold to UV-irradiation. In order to investigation whether the increase of UV130 transcripts was a specific results of UV-irradiation, UV130 transcript levels were examined after treating the cells to Methylmethane sulfonate (MMS). The transcripts of UV130 were not induced by treatment of 0.25% MMS. These results implied that the effects of damaging agents are complex and different regulatory pathways exist for the induction of these genes. To characterize the structure of UV130 gene, nucleotide sequences were analyzed. The nucleotide sequence of 1,340 nucleotide excluding poly(A) tail contains one open reading frame, which encodes a protein of 270 amino acids. The predicted amino acid sequences of UV130 do not exhibit any significant similarity to ther known sequences in the database.

• Korean Institute of Interior Design Journal
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• v.17 no.6
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• pp.120-127
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• 2008

Korean space design image is a kind of writing as well as one of the sign that dissembles itself as a direct transcript of what it represents. Moreover it is pictorial representation and notions such as mental and perceptual imaginary. Significance of Iconology lies in how we transform image and the imagination that produces it into power of trust and respect. From this point of view, the process of this study is illustrated as follows: At first, this study search out concept of archetype, collective unconsciousness and collective representation that found principles on basic theory for interpretation of korean space icon. Secondly, it mentions theoretical background of iconological contents and structure. And it clarifies Iconology as a method that is applicable logic for Korean space design. Finally, as an analysis of korean space design, this study analyse in three steps that are pre-iconological description, iconological analysis, iconological interpretation each in terms of modernization at regional korean space design. In the step of the pre-iconological description, it describe visual representative style based on era and place. In the step of the iconological analysis, the typical structure is classified in status, vernacular, ethnic, traditional. In the step of the iconological interpretation, connotation is categorized into allegory, multivalence, potential. Through this process, this study suggest that iconology is an appropriate analysis system of Korean space design images that represent symbols combined with our collective emotion.