• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1997년도 춘계학술대회
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• pp.81-81
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• 1997

We previously demonstrated an enhancer-like positive regulatory element within a 259-bp sequence (-2352 to-2094 bp) of the human CYP1A2 gene in HepG2 cells. Three protein binding sites were identified by DNase I footprint analyses within the 259-bp sequence: protected region A PRA ( -2283 to-2243 bp), PRB (-2218 to-2187 bp), and PRC (-2124 to-2098 bp) (I. Chung and E. Bresnick, Mol. Pharmacol. 47, 677-685, 1995). In the present study, the functional significance of those protected regions was examined. Transfection experiments with deletion and substitution mutants defined the PRB and PRC as containing positive and negative regulatory elements, respectively. Human breast carcinoma MCF-7 cells were cotransfected with a hepatocyte nuclear factor-1 (HNF-1) expression vector and CYP1A2 promoter-or thymidine kinase promoter-luciferase remoter gene constructs. HNF-1, which contributes to the liver specificity of genes, enhanced reporter gene activity in a PRC sequence-dependent manner. These results suggested that PRC could exist bound to a repressor which was displaceable by other transcription factors such as HNF-1. Results obtained by transfection of HepG2 hepatoma cells with various PRB substitution mutant-luciferase gene fusion constructs indicated that the entire sequence of PRB was necessary for promoter activity. Consequently, the regulation of CYP1A3 expression is very complex, requiring a number of both positive and negative regulatory factors.

• Journal of Microbiology and Biotechnology
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• 제24권10호
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• pp.1301-1307
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• 2014

White-rot fungi of the genus Bjerkandera are cosmopolitan and have shown potential for industrial application and bioremediation. When distinguishing morphological characters are no longer present (e.g., cultures or dried specimen fragments), characterizing true sequences of Bjerkandera is crucial for accurate identification and application of the species. To build a framework for molecular identification of Bjerkandera, we carefully identified specimens of B. adusta and B. fumosa from Korea based on morphological characters, followed by sequencing the internal transcribed spacer region and 28S nuclear ribosomal large subunit. The phylogenetic analysis of Korean Bjerkandera specimens showed clear genetic differentiation between the two species. Using this phylogeny as a framework, we examined the identification accuracy of sequences available in GenBank. Analyses revealed that many Bjerkandera sequences in the database are either misidentified or unidentified. This study provides robust reference sequences for sequence-based identification of Bjerkandera, and further demonstrates the presence and dangers of incorrect sequences in GenBank.

• 한국복합재료학회:학술대회논문집
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• 한국복합재료학회 2002년도 추계학술발표대회 논문집
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• pp.233-236
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• 2002

An efficient method was developed in this study to obtain optimal stacking sequences, thicknesses, and minimum weights of stiffened laminated composite shells under combined loading conditions and stiffener layouts using genetic algorithms (GAs) and finite element analyses. Among many parameters in designing composite laminates determining a optimal stacking sequence that may be formulated as an integer programming problem is a primary concern. Of many optimization algorithms, GAs are powerful methodology for the problem with discrete variables. In this paper the optimal stacking sequence was determined, which gives the maximum critical buckling load factor and the minimum weight as well. To solve this problem, both the finite element analysis by ABAQUS and the GA-based optimization procedure have been implemented together with an interface code. Throughout many parametric studies using this analysis tool, the influences of stiffener sizes and three different types of stiffener layouts on the stacking sequence changes were throughly investigated subjected to various combined loading conditions.

• 한국기계가공학회지
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• 제15권6호
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• pp.24-34
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• 2016

To attach a fuel tank to an excavator, two sets of mounting plates on which three bosses are attached are welded onto the tank. In this study, the welding process of a fuel tank for an excavator was modeled using a finite element numerical method. The tank was modeled as a simple plate to which the mounting plate or bosses were attached by fillet welding. Thermal and thermo-elasto-plastic analyses of the welding process were carried out to predict the temperature distribution and material distortion during welding, respectively. Three different welding sequences for the tank were also modelled to compare the deformation that occurred due to each welding sequence. The results of the analysis predicted that changing the welding sequence around the mounting plate could not position the boss within the allowable dimensional range. The results also revealed the sequence in which the maximum distortion of the bosses welded onto the tank was 30% less than the maximum distortion due to the other sequences.

• BMB Reports
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• 제39권6호
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• pp.717-721
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• 2006

In vitro analyses of type I signal peptidase activities require protein precursors as substrates. Usually, these pre-proteins are expressed in vitro and cleavage of the signal sequence is followed by SDS polyacrylamide gel electrophoresis coupled with autoradiography. Radioactive amino acids have to be incorporated in the expressed protein, since the amount of the in vitro expressed protein is usually very low and processing of the signal peptide cannot be followed by SDS polyacrylamide gel electrophoresis alone. Here we describe a rapid and simple method to express large amounts of a protein precursor in E. coli. We have analyzed the effect of ionophors as well as of azide on the accumulation of expressed protein precursors. Azide blocks the function of SecA and the ionophors dissipate the electrochemical gradient across the cytoplasmic membrane of E. coli. Addition of azide ions resulted in the formation of inclusion bodies, highly enriched with pre-apo-plastocyanine. Plastocyanine is a soluble copper protein, which can be found in the periplasmic space of cyanobacteria as well as in the thylakoid lumen of cyanobacteria and chloroplasts, and the pre-protein contains a cleavable signal sequence at its N-terminus. After purification of cyanobacterial pre-apo-plastocyanine, its signal sequence can be cleaved off by the E. coli signal peptidase, and protein processing was followed on Coomassie stained SDS polyacrylamide gels. We are optimistic that the presented method can be further developed and applied.

• Journal of Ginseng Research
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• 제35권1호
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• pp.21-30
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• 2011

Panax ginseng root has been used as a major source of ginsenoside throughout the history of oriental medicine. In recent years, scientists have found that all of its biomass, including embryogenic calli and flower buds can contain similar active ingredients with pharmacological functions. In this study, transcriptome analyses were used to identify different gene expressions from embryogenic calli and fl ower buds. In total, 6,226 expressed sequence tags (ESTs) were obtained from cDNA libraries of P. ginseng. Insilico analysis was conducted to annotate the putative sequences using gene ontology functional analysis, Kyoto Encyclopedia of Genes and Genomes orthology biochemical analysis, and interproscan protein functional domain analysis. From the obtained results, genes responsible for growth, pathogenicity, pigments, ginsenoside pathway, and development were discussed. Almost 83.3% of the EST sequence was annotated using one-dimensional insilico analysis.

• 한국소음진동공학회논문집
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• 제18권12호
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• pp.1262-1269
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• 2008

Though multilayered foam-panel structures have been widely used to reduce sound transmission in various fields, most of the previous works to design them were conducted by repeated analyses or experiments based on initially given configurations or sequences. Therefore, it was difficult to obtain an optimal sequence of multilayered foam-panel structure yielding superior sound isolation capability. In this work, we propose a new design method to sequence a multi-panel structure lined with a poroelastic material having maximized sound transmission loss. Being formulated as a one-dimensional topology optimization problem fur a given target frequency, the optimal sequencing of panel-poroelastic layers is systematically carried out in an iterative manner. In this method, a panel layer is expressed as a limiting case of a poroelastic layer to facilitate the optimization process. This means that main material properties of a poroelastic material are treated as interpolated functions of design variable. The designed sequences of panel-poroelastic multilayer were shown to be significantly affected by the target frequencies; more panels were obtained at higher target frequency. The sound transmission loss of the system was calculated by the transfer matrix derived from Biot's theory.

• Parasites, Hosts and Diseases
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• 제56권2호
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• pp.153-165
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• 2018

Development of an effective vaccine is critically needed for the prevention of malaria. One of the key antigens for malaria vaccines is the apical membrane antigen 1 (AMA-1) of the human malaria parasite Plasmodium falciparum, the surface protein for erythrocyte invasion of the parasite. The gene encoding AMA-1 has been sequenced from populations of P. falciparum worldwide, but the haplotype diversity of the gene in P. falciparum populations in the Greater Mekong Subregion (GMS), including Thailand, remains to be characterized. In the present study, the AMA-1 gene was PCR amplified and sequenced from the genomic DNA of 65 P. falciparum isolates from 5 endemic areas in Thailand. The nearly full-length 1,848 nucleotide sequence of AMA-1 was subjected to molecular analyses, including nucleotide sequence diversity, haplotype diversity and deduced amino acid sequence diversity and neutrality tests. Phylogenetic analysis and pair-wise population differentiation ($F_{st}$ indices) were performed to infer the population structure. The analyses identified 60 single nucleotide polymorphic loci, predominately located in domain I of AMA-1. A total of 31 unique AMA-1 haplotypes were identified, which included 11 novel ones. The phylogenetic tree of the AMA-1 haplotypes revealed multiple clades of AMA-1, each of which contained parasites of multiple geographical origins, consistent with the $F_{st}$ indices indicating genetic homogeneity or gene flow among geographically distinct populations of P. falciparum in Thailand's borders with Myanmar, Laos and Cambodia. In summary, the study revealed novel haplotypes and population structure needed for the further advancement of AMA-1-based malaria vaccines in the GMS.

• 생명과학회지
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• 제14권4호
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• pp.593-607
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• 2004

유해성 적조생물 Cochlodinium polykrikoides/Gyrodinium aurelum을 포함한 43 종류의 와편조류를 대상으로 SSU 부위 유전자를 분석했다. 유전자 염기서열에 의거한 상호 계통수는 parisomny, distance, maxium 방법으로 실행했다. Noctilura scintillans, Gonyaulax spinifera와 Crythecodinium cohnii 종들은 와편모조류 중 가장 유전적으로 먼 것으로 보였다. Alexandrium과 Symbiodinium 종간의 bootstrap는 70% 이상의 상호 단일 계통도를 보인 반면에, Gymnodinium과 Gyrodinium은 근립절약계수와 최대 유사도 방법에서 다형 계통도를 나타내었다. Gyroddinium aurelum과 G. dorsum/G. galathenum의 유전적 분화율은 7.4% (45 bp) 였고, G. aurelum과 G. mikimotoi 상호간에는 0.9% (5bp) 밖에 나타나지 않았다. 또한 C. polykrikoides와 G. aurelum도 5.2% (31bp)로 낮은 유전적 분화율을 보였다. 계통도를 분석한 결과 G. aureolum과 C. polykrikoides는 Gyrodinium 보다 Gymnodinim 속에 훨씬 더 근접하게 나타났다.

• Journal of Applied Biological Chemistry
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• 제59권4호
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• pp.299-304
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• 2016

PCR 산물을 이용한 시퀀싱방법 중 Illumina 플랫폼으로 시퀀싱을 수행하면 100개 이상의 인위적인 시퀀스가 생겨나며, 그러한 인위적으로 형성되는 시퀀스에 의해 Operational taxonomic units를 기반으로 한 미생물생태 변화 및 네트워크 분석에 영향을 미친다. 이러한 문제점이 있음에도 불구하고 분변미생물생태를 분석하는데 Illumina에서 제공하고 있는 시퀀싱을 주된 방법으로 사용하고 있으며, 또한 그러한 시퀀스 기반의 분변미생물 생태분석 결과는 분변샘플상태(i.e., 분변 보관 기간, 분변양, 분변의 신선도)에 따라 상이하게 나타난다. 본 연구에서는 분변샘플의 동결건조가 시퀀스 데이터의 퀄리티를 향상시키는지 관해 조사하였으며, 이를 통해 분변샘플에 동결건조처리는 전체적인 미생물생태구조를 변화시키지는 않지만 인위적으로 형성되었을 가능성이 있는 시퀀스의 수를 감소시키는 것으로 확인되었다. 따라서, 분변으로부터 DNA를 추출하기 이전에 동결건조처리하는 방법을 Illumina 기반의 분변미생물생태분석에 사용하는 것을 권장한다.