• BMB Reports
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• v.38 no.4
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• pp.474-480
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• 2005

Curcumin, a major active component of turmeric, has been identified as an inhibitor of the transcriptional activity of activator protein-1 (AP-1). Recently, it was also found that curcumin and synthetic curcumin derivatives can inhibit the binding of Jun-Fos, which are the members of the AP-1 family, to DNA. However, the mechanism of this inhibition by curcumin and its derivatives was not disclosed. Since the binding of Jun-Fos dimer to DNA can be modulated by redox control involving conserved cysteine residues, we studied whether curcumin and its derivatives inhibit Jun-Fos DNA binding activity via these residues. However, the inhibitory mechanism of curcumin and its derivatives, unlike that of other Jun-Fos inhibitors, was found to be independent of these conserved cysteine residues. In addition, we investigated whether curcumin derivatives can inhibit AP-1 transcriptional activity in vivo using a luciferase assay. We found that, among the curcumin derivatives examined, only inhibitors shown to inhibit the binding of Jun-Fos to DNA by Electrophoretic Mobility Shift Assay (EMSA) inhibited AP-1 transcriptional activity in vivo. Moreover, RT-PCR revealed that curcumin derivatives, like curcumin, downregulated c-jun mRNA in JB6 cells. These results suggest that the suppression of the formation of DNA-Jun-Fos complex is the main cause of reduced AP-1 transcriptional activity by curcuminoids, and that EMSA is a suitable tool for identifying inhibitors of transcriptional activation.

• Journal of Life Science
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• v.11 no.5
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• pp.464-469
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• 2001

A gene, dnaK2, encoding a distinct member of the HSP70 family of molecular chaperones is isolated from the halotolerant cyanobactrium Aphanothece halophytica. The dnak2 gene encodes a molecular wight of 68 kDa polypeptide with predicted 616 amino acid residues. The DnaK2 protein has a structural characteristic of bacterial DnaK homologues and shows high similarity to other HSP70/Dank proteins. The danK2 transcripts are hardly detectable at 28$^{\circ}C$ and strongly induced upon heat stress. It is also found that dnaK2 transcript is increased by high-salinity stress even in the absence of heat stress. These results suggest that the DnaK2 protein plays an important role in protecting A. halophytica against damage caused by salt stress at well as heat stress.

• Journal of Microbiology and Biotechnology
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• v.31 no.9
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• pp.1323-1329
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• 2021

Micro-scale magnetic beads are widely used for isolation of proteins, DNA, and cells, leading to the development of in vitro diagnostics. Efficient isolation of target biomolecules is one of the keys to developing a simple and rapid point-of-care diagnostic. A zinc finger protein (ZFP) is a double-stranded (ds) DNA-binding domain, providing a useful scaffold for direct reading of the sequence information. Here, we utilized two engineered ZFPs (Stx2-268 and SEB-435) to detect the Shiga toxin (stx2) gene and the staphylococcal enterotoxin B (seb) gene present in foodborne pathogens, Escherichia coli O157 and Staphylococcus aureus, respectively. Engineered ZFPs are immobilized on a paramagnetic bead as a detection platform to efficiently isolate the target dsDNA-ZFP bound complex. The small paramagnetic beads provide a high surface area to volume ratio, allowing more ZFPs to be immobilized on the beads, which leads to increased target DNA detection. The fluorescence signal was measured upon ZFP binding to fluorophore-labeled target dsDNA. In this study, our system provided a detection limit of ≤ 60 fmol and demonstrated high specificity with multiplexing capability, suggesting a potential for development into a simple and reliable diagnostic for detecting multiple pathogens without target amplification.

• Biomolecules & Therapeutics
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• v.3 no.4
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• pp.311-315
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• 1995

The K. aerogenes ureal gene product was previously shown to facilitate assembly of the crease metallocenter (Lee, M. H., Mulrooney, S. B., Renner, M. J., Markowicz, Y., and Hausinger, R. P. (1992) J. Bacteriol. 174, 4324-4330). UreG protein has now been purified and characterized. Although the protein is predicted to possess a putative NTP-binding P-loop motif, equilibrium dialysis studies showed negative results. Immunogold electron microscopic studies using polyclonal antibodies directed against UreG protein confirm that UreG is located in the cytoplasm as predicted in the DNA sequence.

• BMB Reports
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• v.34 no.5
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• pp.452-456
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• 2001

Surface plasmon resonance has been used for a biospecific interaction analysis between two macromolecules in real time. Determination of an antibody that is capable of specifically interacting with the native form of antigen is very useful for many biological and medical applications. Twenty monoclonal antibodies against the $\alpha$ subunit of E. coli DNA polymerase III were screened for specifically recognizing the native form of protein using surface plasmon resonance. Only four monoclonal antibodies among them specifically recognized the native $\alpha$ protein, although all of the antibodies were able to specifically interact with the denatured $\alpha$ subunit. These antibodies failed to interfere with the interaction between the $\tau$ and $\alpha$ subunits that were required for dimerization of the two polymerases at the DNA replication fork. This real-time analysis using surface plasmon resonance provides an easy method to screen antibodies that are capable of binding to the native form of the antigen molecule and determine the biological interaction between the two molecules.

• Korean Journal of Environmental Biology
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• v.38 no.3
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• pp.350-354
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• 2020

Many parrots are considered endangered species due to threats from human activities. Gender determination is of great importance for biological studies and the conservation of endangered parrots. However, like other birds, gender determination in parrots is hindered due to the lack of external dimorphism between males and females. A molecular approach using the chromo-helicase-DNA binding protein 1 (CHD1) gene is commonly used for sexing birds. This study aimed to determine the gender of parrots from Korean zoos based on amplification and visualization of the partial CHD1 gene. The samples of 13 parrot species were collected from three different zoos in Korea and the extracted DNA templates were amplified using CHD1 gene primers. The gender of 27 samples of 13 species was determined by visualizing the PCR products on an agarose gel. While male parrots were indicated by a single band, female parrots were indicated by double bands. The findings provide additional information, which might be helpful for the management and care of parrots in Korean zoos.

• The Korean Journal of Physiology and Pharmacology
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• v.14 no.4
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• pp.229-233
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• 2010

Amyloid precursor protein binding protein-1 (APP-BP1) binds to the carboxyl terminus of amyloid precursor protein and serves as a bipartite activation enzyme for the ubiquitin-like protein, NEDD8. Previously, it has been reported that APP-BP1 rescues the cell cycle S-M checkpoint defect in Ts41 hamster cells, that this rescue is dependent on the interaction of APP-BP1 with hUba3. The exogenous expression of APP-BP1 in neurons has been reported to cause DNA synthesis and apoptosis via a signaling pathway that is dependent on APP-BP1 binding to APP. These results suggest that APP-BP1 overexpression contributes to neurodegeneration. In the present study, we explored whether APP-BP1 expression was altered in the brains of Tg2576 mice, which is an animal model of Alzheimer's disease. APP-BP1 was found to be up-regulated in the hippocampus and cortex of 12 month-old Tg2576 mice compared to age-matched wild-type mice. In addition, APP-BP1 knockdown by siRNA treatment reduced cullin-1 neddylation in fetal neural stem cells, suggesting that APP-BP1 plays a role in cell cycle progression in the cells. Collectively, these results suggest that increased expression of APP-BP1, which has a role in cell cycle progression in neuronal cells, contributes to the pathogenesis of Alzheimer's disease.

• Korean journal of applied entomology
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• v.34 no.2
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• pp.100-105
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• 1995

A whole body extract (WBE), a crude nuclear fraction, of aphids was prepared and used to identify the proteins which bound specifically to 5'-upstream regions of the transcription initiation site of the aphid ribosomal RNA gene (rDNA). While DNA fragment (-263/-195) was bound by only one specific 53 kDa protein, two DNA fragments, A(-194/23) and B(-393/-264), were commonly bound by three proteins (52 kDa, 50 kDa and 40 kDa). It was also revealed that the formation of he DNA-protein complex requires a cation.

• Proceedings of the Korean Information Science Society Conference
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• 2004.04b
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• pp.307-309
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• 2004

급성 심근경색 진단용 DNA 컴퓨팅 시스템 모듈로서, 트로포닌 I (troponin I, Tnl). 트로포닌 T (troponin T, TnT). 미오글로빈 (myoglobin), C-반응 단백질 (C-reactive protein, CRP) 과 각각 결합할 수 있는 네 가지 종류의 앱타머틀 선정하고, 이의 개발을 시도하여, 그 중 첫 번째로 C-반응 단백질-결합 앱타머를 SELEX 기법을 이용하여 선별해내었다. 또한, 선별된 앱타머 염기서열에 기초하여 각각 10-mer 길이의 FDNA 와 QDNA 를 제작하고, 표적 단백질 (CRP) 과 혼합시켜 형광발현 변화의 추이를 살펴보았다. 앱타머 및 FDNA. QDNA 가 결합할 경우에는 형광감쇄효과가 발생하므로, 형광감쇄효과가 일어나지 않은 경우에 비하여 현저하게 형광측정값이 저조하게 나타나는 현상을 확인할 수 있었다. 향후 연구로, 나머지 세 가지 종류의 앱타머를 SELEX기법을 이용하여 선별해내고. 기확보된 C-반응 단백질-결합 앱타머 모듈과 함께 논리회로를 구성하는 DNA 컴퓨팅 칩을 제작할 예정이다.

• Korean Journal of Plant Tissue Culture
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• v.24 no.1
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• pp.1-35
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• 1997

Fertilized egg, by successive cell divisions, differentiates into different tissues and organs with various structures and functions. Different cells and tissues contain different proteins, products of selective gene expression. Not all the genes in any genomes are equally active, temporal and spatial gene expression being the general rule. Present paper attempts to review the tanscriptional mechanisms or the initiations of transcription from several angles. In some of the organisms the genes in the process of transcription or the genes in the inactive state can be seen under the light microscope. Some bands of Drosophila polytene chromosomes may exhibit a swollen or puff appearance under certain conditions. A puff, unfolded or decondensed form of chromomere, represents sets of intense transcriptional activity or RNA synthesis. The heterochromatic X chromosome whose genes remain inactive in the female mammals can be visualized as a dark staining structure called Barr body, Configuration of chromatin differs between transcribed and nontranscribed chromatin. Modification to the chromatin facilitates RNA synthesis. The movement of large polymerase molecule along the DNA would probably be facilitated if some modifications of the chromatin configuration is effected. Methylation of cytosines in CG sequences is associated with inactive genes. Methylation can play a role in determination of mammalian cells during embryogenesis. Demethylation is necessary for the gene to be expressed during development A histone modification that is also known to be correlated with transcriptional capacity of chromatin is acetylation of the lysine residues of the core histones. Chromatin containing a high level of histone acetylation is very sensitive to DNase 1. For the transcription to occur TBP must first bind to the TATA box. Another TF, TF IIB, then binds to the promoter-TBP complex, facilitating the access of RNA polymerase to the transcription initiation site. As recently as eight years ago researchers assumed that histones were irrelevant to the regulation of gene expression. Histones combine with the DNA to form nucleosome of the chromatin. Histones are vital participant in gene regulation. Histone and basal factors compete for access to TATA box. When DNA is exposed to basal factors before histones are introduced, the basal factors assemble on TATA boxes preventing the access of histones, allowing transcription to occur, for transcription to begin, activator protein at the upstream activation sequence or enhancer must interact with the tail of histone H4 at TATA box and cause the histone role particle to dissociate from the TATA box leading to partial breakup of the histone core particle and allowing the basal factors to bind to the TATA box. New concept of genomic flux in contrast to the old concept of static genome has been developed based on the powerful new molecular techniques. Genomic changes such as repetitive DNAs and transposable elements, it is assumed but not yet proved, may affect some of the developmental patterns that characterize particular cells, tissues, organs, and organisms. In the last decade or so remarkable achievement have been made in the researches of the structures and functions of TFs and the specific target sequences located in promoters or enhancers where these TFs bind. TFs have independent domains that bind DNA and that activate transcription. DNA binding domain of TFs serves to bring the protein into the right location. There are many types of DNA binding domains. Common types of motifs can be found that are responsible for binding to DNA. The motifs are usually quite short and comprise only a small part of the protein structure. Steroid receptors have domains for hormone binding, DNA binding, and activating transcription. The zinc finger motif comprises a DNA binding domain. Leucine zipper consist of a stretch of amino acids with a leucine residue in every seventh position Two proteins form a dimer because they interact by means of leucine zippers on similar α-helical domain. This positions their DNA binding basic domains for interaction with the two halves of a DNA sequence with dyad symmetry of TGACTCA, ACTGAGT.