• Title/Summary/Keyword: FRET

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The Synthesis and Optical Properties of Silica Coated CdSe/ZnS QDs (실리카가 코팅된 양자점의 코팅두께에 따른 광 특성 변화)

  • Lee, Ji-Hye;Shin, Hyun Ho;Lee, Jong-Heun;Hyun, Sang Il;Koo, Eunhae
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.26 no.3
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    • pp.221-226
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    • 2013
  • The water soluble quantum dots (QDs) are synthesized by the phase transfer and silica coating reaction. The photoluminescence intensity of silica-coated QDs are mainly affected by the amount of phase transfer agent, SDS (sodium dodecyl sulfate), and the maximum value is obtained at the cmc (critical micell concentration) concentration of SDS in the phase transfer reaction. Based on fluorescence spectra and field emission transmission electron microscope (FETEM), the energy transfer rate by forster resonance energy transfer (FRET) is increasing with the thickness of the silica shell coated on CdSe/ZnS QDs.

Pepper Blight Disease Inhibition Metagenome Clone Screening Using Soil Metagenome Library (토양 Metagenome Library로부터 고추역병 저해 클론 탐색)

  • Park, Hae-Chul;Sung, So-Ra;Kim, Dong-Gwan;Koo, Bon-Sung;Jeong, Byeong-Moon;Kim, Jin-Heung;Yoon, Moon-Young
    • Korean Journal of Microbiology
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    • v.45 no.2
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    • pp.228-231
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    • 2009
  • We have purified Phytophthora capsici alpha and beta tubulin from Escherchia coli BL21(DE3). The recombinant alpha and beta tubulins were assembled into microtubule in vitro with specific conditions. The metagenome library was isolated from soil in the Mt. Yeo-Ki, Suwon, Korea and manufactured with the method mentioned in experiment contents for in vitro screening of microtubule assembly screening. FRET effect was used for microtubule assembly inhibitor screening with metagenome library. We got 2 metagenome clones from in vitro screening, and these 2 hit clones showed P. capsici growth inhibition activity on the growing pepper plants. These results suggest that new development of potent inhibitor for pepper blight disease and new approach to prevention of pepper blight disease.

High-throughput SNP Genotyping by Melting Curve Analysis for Resistance to Southern Root-knot Nematode and Frogeye Leaf Spot in Soybean

  • Ha, Bo-Keun;Boerma, H. Roger
    • Journal of Crop Science and Biotechnology
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    • v.11 no.2
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    • pp.91-100
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    • 2008
  • Melting curve analysis of fluorescently labeled DNA fragments is used extensively for genotyping single nucleotide polymorphism(SNP). Here, we evaluated a SNP genotyping method by melting curve analysis with the two probe chemistries in a 384-well plate format on a Roche LightCycler 480. The HybProbe chemistry is based on the fluorescence resonance energy transfer(FRET) and the SimpleProbe chemistry uses a terminal self-quenching fluorophore. We evaluated FRET HybProbes and SimpleProbes for two SNP sites closely linked to two quantitative trait loci(QTL) for southern root-knot nematode resistance. These probes were used to genotype the two parents and 94 $F_2$ plants from the cross of PI 96354$\times$Bossier. The SNP genotypes of all samples determined by the LightCycler software agreed with previously determined SSR genotypes and the SNP genotypes determined on a Luminex 100 flow cytometry instrument. Multiplexed HybProbes for the two SNPs showed a 98.4% success rate and 100% concordance between repeats two of the same 96 DNA samples. Also, we developed a HybProbe assay for the Rcs3 gene conditioning broad resistance to the frogeye leaf spot(FLS) disease. The LightCycler 480 provides rapid PCR on 384-well plate and allows simultaneous amplification and analysis in approximately 2 hours without any additional steps after amplification. This allowed for a reduction of the potential contamination of PCR products, simplicity, and enablement of a streamlined workflow. The melting curve analysis on the LightCycler 480 provided high-throughput and rapid SNP genotyping and appears highly effective for marker-assisted selection in soybean.

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Numerical Analysis of Sunroof Buffeting using STAR-CCM+ (STAR-CCM+를 이용한 썬루프 버페팅 유동 소음 해석)

  • Bonthu, Satish Kumar;Mendonca, Fred;Kim, Ghuiyeon;Back, Young-R.
    • Transactions of the Korean Society for Noise and Vibration Engineering
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    • v.24 no.3
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    • pp.213-218
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    • 2014
  • CFD flow simulation of vehicles with open sunroof and passenger window help the automotive OEM(original equipment manufacturer) to identify the low frequency noise levels in the cabin. The lock-in and lock-off phenomena observed in the experimental studies of sunroof buffeting is well predicted by CFD speed sweep calculations over the operating speed range of the vehicle. The trend of the shear layer oscillation frequency with vehicle speed is also well predicted. The peak SPL from the CFD calculation has a good compromise with the experimental value after incorporating the real world effects into the CFD model by means of artificial compressibility and damping correction. The entire process right from modeling to flow analysis as well as acoustic analysis has been performed within the single environment i.e., STAR-CCM+.

Plasmonic Nanosheet towards Biosensing Applications

  • Tamada, Kaoru
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.02a
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    • pp.105-106
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    • 2013
  • Surface plasmon resonance (SPR) is classified into the propagating surface plasmon (PSP) excited on flat metal surfaces and the local surface plasmon (LSP) excited by metalnanoparticles. It is known that fluorescence signals are enhanced by these two SPR-fields.On the other hand, fluorescence is quenched by the energy transfer to metal (FRET). Bothphenomena are controlled by the distance between dyes and metals, and the degree offluorescence enhancement is determined by the correlation. In this study, we determined thecondition to achieve the maximum fluorescence enhancement by adjusting the distance of ametal nanoparticle 2D sheet and a quantum dots 2D sheet by the use of $SiO_2$ spacer layers. The 2D sheets consisting of myristate-capped Ag nanoparticles (AgMy nanosheets) wereprepared at the air-water interface and transferred onto hydrophobized gold thin films basedon the Langmuir-Schaefer (LS) method [1]. The $SiO_2$ sputtered films with different thickness (0~100 nm) were deposited on the AgMy nanosheet as an insulator. TOPO-cappedCdSe/CdZnS/ZnS quantum dots (QDs, ${\lambda}Ex=638nm$) [2] were also transferred onto the $SiO_2$ films by the LS method. The layered structure is schematically shown in Fig. 1. The result of fluorescence measurement is shown in Fig. 2. Without the $SiO_2$ layer, the fluorescence intensity of the layered QD film was lower than that of the original QDs layer, i.e., the quenching by FRET was predominant. When the $SiO_2$ thickness was increased, the fluorescence intensity of the layered QD film was higher than that of the original QDs layer, i.e., the SPR enhancement was predominant. The fluorescence intensity was maximal at the $SiO_2$ thickness of 20 nm, particularly when the LSPR absorption wavelength (${\lambda}=480nm$) was utilized for the excitation. This plasmonic nanosheet can be integrated intogreen or bio-devices as the creation point ofenhanced LSPR field.

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Photoluminescent Graphene Oxide Microarray for Multiplex Heavy Metal Ion Analysis

  • Liu, Fei;Ha, Hyun Dong;Han, Dong Ju;Park, Min Su;Seo, Tae Seok
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.08a
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    • pp.281.2-281.2
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    • 2013
  • Since heavy metal ions included in water or food resources have critical effects on human health, highly sensitive, rapid and selective analysis for heavy metal detection has been extensively explored by means of electrochemical, optical and colorimetric methods. For example, quantum dots (QDs), such as semiconductor QDs, have received enormous attention due to extraordinary optical properties including high fluorescence intensity and its narrow emission peaks, and have been utilized for heavy metal ion detection. However, the semiconductor QDs have a drawback of serious toxicity derived from cadmium, lead and other lethal elements, thereby limiting its application in the environmental screening system. On the other hand, Graphene oxide (GO) has proven its superlative properties of biocompatibility, unique photoluminescence (PL), good quenching efficiency and facile surface modification. Recently, the size of GO was controlled to a few nanometers, enhancing its optical properties to be applied for biological or chemical sensors. Interestingly, the presence of various oxygenous functional groups of GO contributes to opening the band gap of graphene, resulting in a unique PL emission pattern, and the control of the sp2 domain in the sp3 matrix of GO can tune the PL intensity as well as the PL emission wavelength. Herein, we reported a photoluminescent GO array on which heavy metal ion-specific DNA aptamers were immobilized, and sensitive and multiplex heavy metal ion detection was performed utilizing fluorescence resonance energy transfer (FRET) between the photoluminescent monolayered GO and the captured metal ion.

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Force Transmission in Cellular Adherens Junction Visualized by Engineered FRET Alpha-catenin Sensor (형광공명에너지전이 알파카테닌 센서를 활용한 세포 부착접합부에서의 힘 전달 이미징)

  • Jang, Yoon-Kwan;Suh, Jung-Soo;Suk, Myungeun;Kim, Tae-Jin
    • Korean Chemical Engineering Research
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    • v.59 no.3
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    • pp.366-372
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    • 2021
  • Cadherin-Catenin complex is thought to play an essential role in the transmission of force at adherens junction. Due to the lack of proper tools to visualize and detect mechanical force signals, the underlying mechanism by which the cadherin-catenin complex regulates force transmission at intercellular junctions remains elusive. In this study, we visualize cadherin-mediated force transmission using an engineered α-Catenin sensor based on fluorescence resonance energy transfer. Our results reveal that α-catenin is a key force transducer in cadherin-mediated mechanotransduction at cell-cell junctions. Thus, our finding will provide important insights for studying the effects of chemical and physical signals on cell-cell communication and the relationship between physiological and pathological phenomena.

Ginsenoside F2 Restrains Hepatic Steatosis and Inflammation by Altering the Binding Affinity of Liver X Receptor Coregulators

  • Kyurae Kim;Myung-Ho Kim;Ji In Kang;Jong-In Baek;Byeong-Min Jeon;Ho Min Kim;Sun-Chang Kim;Won-Il Jeong
    • Journal of Ginseng Research
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    • v.48 no.1
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    • pp.89-97
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    • 2024
  • Background: Ginsenoside F2 (GF2), the protopanaxadiol-type constituent in Panax ginseng, has been reported to attenuate metabolic dysfunction-associated steatotic liver disease (MASLD). However, the mechanism of action is not fully understood. Here, this study investigates the molecular mechanism by which GF2 regulates MASLD progression through liver X receptor (LXR). Methods: To demonstrate the effect of GF2 on LXR activity, computational modeling of protein-ligand binding, Time-resolved fluorescence resonance energy transfer (TR-FRET) assay for LXR cofactor recruitment, and luciferase reporter assay were performed. LXR agonist T0901317 was used for LXR activation in hepatocytes and macrophages. MASLD was induced by high-fat diet (HFD) feeding with or without GF2 administration in WT and LXRα-/- mice. Results: Computational modeling showed that GF2 had a high affinity with LXRα. LXRE-luciferase reporter assay with amino acid substitution at the predicted ligand binding site revealed that the S264 residue of LXRα was the crucial interaction site of GF2. TR-FRET assay demonstrated that GF2 suppressed LXRα activity by favoring the binding of corepressors to LXRα while inhibiting the accessibility of coactivators. In vitro, GF2 treatments reduced T0901317-induced fat accumulation and pro-inflammatory cytokine expression in hepatocytes and macrophages, respectively. Consistently, GF2 administration ameliorated hepatic steatohepatitis and improved glucose or insulin tolerance in WT but not in LXRα-/- mice. Conclusion: GF2 alters the binding affinities of LXRα coregulators, thereby interrupting hepatic steatosis and inflammation in macrophages. Therefore, we propose that GF2 might be a potential therapeutic agent for the intervention in patients with MASLD.

The Effect of the Oxygen Scavenging System on the pH of Buffered Sample Solutions: in the Context of Single-molecule Fluorescence Measurements

  • Kim, Sung-Eun;Lee, Il-Buem;Hong, Seok-Cheol
    • Bulletin of the Korean Chemical Society
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    • v.33 no.3
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    • pp.958-962
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    • 2012
  • In single-molecule fluorescence experiment, the oxygen scavenging system is indispensable for avoiding photo-bleaching of fluorescent dyes. Here we report that the gloxy-based oxygen scavenging system commonly used in single molecule fluorescence experiments can disturb the solution pH considerably. To track in situ pH change, we utilized the pH-sensitive conformational transition of i-motif and examined the transition with ensemble and single-molecule FRET measurements. Based on our results, we also suggested several practical remedies for the stability of the solution pH.