• Title/Summary/Keyword: Single-molecule

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Biosynthesis of a Biological Active Single Chain Equine Chorionic Gonado-tropin

  • Min, Kwan-Sik
    • Journal of Life Science
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    • v.11 no.2
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    • pp.103-107
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    • 2001
  • The equine chorionic gonadotropin (eCG) subunits $\alpha$ and ${\beta}$ are transcribed from different genes and associate noncovalently to form the bioactive eCG heterodimer. Dimerization is rate limiting for eCG secretion, and dissociation leads to hormone inactivation. The correct conformation of the heterodimer is alto important for efficient secretion, hormone-specific post-translational modifications, receptor binding and signal transduction. To determine whether ${\alpha}$ and ${\beta}$ subunits can be synthesized as a single polypeptide chain (tethered-eCG) and also display biological activity, the tethered-eCG molecule by fusing the carboxyl terminus of the eCG ${\beta}$-subunit to the amino terminus of the af-subunit was construe-ted and transfected into chinese hamster ovary (CHO-Kl) cells. LH- and FSH-like activities were assayed in terms of testosterone production and aromatase activity in primary cultured rat Leydig cells and granulosa cells, respectively. The tethered-eCG was efficiently secreted and showed similar LH-like activity to the dimeric eCG ${\alpha}$/${\beta}$ and native eCG. FSH-like activity of the tethered-eCG was also shown similarly in comparison with the native and wild type eCG ${\alpha}$/${\beta}$. Our data for the first time suggest that the tethered-eCG can be expressed efficiently and the produced product by the CHO-K1 cells is fully LH- and FSH-like activities in rat in vitro bioassay system. Our results also suggest that this molecular can imply particular models of FSH-like activity not LH-like activity in the eCG. Taken together, these data indicate that the constructs of tethered molecule will be useful in the study of mutants that affect subunit association and/or secretion.

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Characterization of the Rosellinia necatrix Transcriptome and Genes Related to Pathogenesis by Single-Molecule mRNA Sequencing

  • Kim, Hyeongmin;Lee, Seung Jae;Jo, Ick-Hyun;Lee, Jinsu;Bae, Wonsil;Kim, Hyemin;Won, Kyungho;Hyun, Tae Kyung;Ryu, Hojin
    • The Plant Pathology Journal
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    • v.33 no.4
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    • pp.362-369
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    • 2017
  • White root rot disease, caused by the pathogen Rosellinia necatrix, is one of the world's most devastating plant fungal diseases and affects several commercially important species of fruit trees and crops. Recent global outbreaks of R. necatrix and advances in molecular techniques have both increased interest in this pathogen. However, the lack of information regarding the genomic structure and transcriptome of R. necatrix has been a barrier to the progress of functional genomic research and the control of this harmful pathogen. Here, we identified 10,616 novel full-length transcripts from the filamentous hyphal tissue of R. necatrix (KACC 40445 strain) using PacBio single-molecule sequencing technology. After annotation of the unigene sets, we selected 14 cell cycle-related genes, which are likely either positively or negatively involved in hyphal growth by cell cycle control. The expression of the selected genes was further compared between two strains that displayed different growth rates on nutritional media. Furthermore, we predicted pathogen-related effector genes and cell wall-degrading enzymes from the annotated gene sets. These results provide the most comprehensive transcriptomal resources for R. necatrix, and could facilitate functional genomics and further analyses of this important phytopathogen.

MicroRNA super-resolution imaging in blood for Alzheimer's disease

  • Mirae Lee;Jiwon Woo;Sang Tae Kim;Minho Moon;Sang Yun Kim;Hanna Cho;Sujin Kim;Han-Kyeol Kim;Jeong-Yoon Park
    • BMB Reports
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    • v.56 no.3
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    • pp.190-195
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    • 2023
  • We propose a novel blood biomarker detection method that uses miRNA super-resolution imaging to enable the early diagnosis of Alzheimer's disease (AD). Here, we report a single-molecule detection method for visualizing disease-specific miRNA in tissue from an AD mice model, and peripheral blood mononuclear cells (PBMCs) from AD patients. Using optimized Magnified Analysis of Proteome (MAPs), we confirmed that five miRNAs contribute to neurodegenerative disease in the brain hippocampi of 5XFAD and wild-type mice. We also assessed PBMCs isolated from the whole blood of AD patients and a healthy control group, and subsequently analyzed those samples using miRNA super-resolution imaging. We detected more miR-200a-3p expression in the cornu ammonis 1 and dentate gyrus regions of 3 month-old 5XFAD mice than in wild-type mice. Additionally, miRNA super-resolution imaging of blood provides AD diagnosis platform for studying miRNA regulation inside cells at the single molecule level. Our results present a potential liquid biopsy method that could improve the diagnosis of early stage AD and other diseases.

One-dimensional Nanomaterials for Field Effect Transistor (FET) Type Biosensor Applications

  • Lee, Min-Gun;Lucero, Antonio;Kim, Ji-Young
    • Transactions on Electrical and Electronic Materials
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    • v.13 no.4
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    • pp.165-170
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    • 2012
  • One-dimensional, nanomaterial field effect transistors (FET) are promising sensors for bio-molecule detection applications. In this paper, we review fabrication and characteristics of 1-D nanomaterial FET type biosensors. Materials such as single wall carbon nanotubes, Si nanowires, metal oxide nanowires and nanotubes, and conducting polymer nanowires have been widely investigated for biosensors, because of their high sensitivity to bio-substances, with some capable of detecting a single biomolecule. In particular, we focus on three important aspects of biosensors: alignment of nanomaterials for biosensors, surface modification of the nanostructures, and electrical detection mechanism of the 1-D nanomaterial sensors.

Nanoparticle plasmonics: from single molecule chemistry to materials science

  • Kim, Ji-Hwan
    • Proceedings of the Korean Vacuum Society Conference
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    • 2015.08a
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    • pp.76.2-76.2
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    • 2015
  • I will present my research group's recent investigation on how the localized plasmon of a nanoparticle interacts with another plasmon, and with nearby molecules. First, I will demonstrate the use of scattering-type scanning near-field microscopy (s-SNOM) to directly visualize the capacitive / conductive coupling in dimeric nanoparticles and heterometallic nanorods. Second, I will talk about the use of gap-plasmons to locally induce photochemical reactions, and to follow chemical kinetics of individual organic molecules using the gap-plasmons. As a last topic, I will talk about the use of near-field coupling between a scanning probe and graphenes to visualize / identify the stacking domains (e. g., ABA versus ABC-type stacking in triple layer) hidden in multilayer graphenes.

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Fluorescence photon counting rate as a function of dye concentration: Effect of dead time of photon detector (색소 농도에 따른 형광 광자의 계수율 : 광자 검출기의 dead time 효과)

  • 고동섭
    • Korean Journal of Optics and Photonics
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    • v.8 no.4
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    • pp.353-355
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    • 1997
  • A single molecule detection system, which consists of confocal fluorescence microscope and single photon counter, has been used to observe the dye concentration dependence of photon counting rate. With increasing concentration, a saturation effect of counting is observed and demonstrated on the basis of the dead time of photon detector. The equations presented here show the relations between the counting rate and some parameters such as probe volume, quantum efficiency of detector, and fluorescence photon number entered onto detector. The signal-to-noise ratio is also discussed briefly.

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Biological Function of Single Chain Equine Chorionic Gonadotiopin Mutants(C-terminal Deletions)

  • ;;;;N.P JarGil
    • Proceedings of the KSAR Conference
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    • 2004.06a
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    • pp.210-210
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    • 2004
  • Equinechorionic gonadotropin(eCG) is a member of the glycoprotein hormone family which includes FSH, hCG, TSH. These hormone family is characterized by a heterodimeric structure composed a common α-subunit noncovalently linked to a hormone specific β-subunit. To determine a and β-subunits can be synthesized as a single polypeptide chain (tethered-eCG) and also display biological activity, the tethered-molecule by fusing the carboxyl terminus of the eCG β-subunit to the amino terminus of the α-subunit was constructed and transfected into chinese hamster ovary (CHO-K1) cells. (omitted)

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Nanoscale-NMR with Nitrogen Vacancy center spins in diamond

  • Lee, Junghyun
    • Journal of the Korean Magnetic Resonance Society
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    • v.24 no.2
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    • pp.59-65
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    • 2020
  • Nitrogen-Vacancy (NV) center in diamond has been an emerging versatile tool for quantum sensing applications. Amongst various applications, nano-scale nuclear magnetic resonance (NMR) using a single or ensemble NV centers has demonstrated promising results, opening possibility of a single molecule NMR for its chemical structural studies or multi-nuclear spin spectroscopy for quantum information science. However, there is a key challenge, which limited the spectral resolution of NMR detection using NV centers; the interrogation duration for NV-NMR detection technique has been limited by the NV sensor spin lifetime (T1 ~ 3ms), which is orders of magnitude shorter than the coherence times of nuclear spins in bulk liquid samples (T2 ~ 1s) or intrinsic 13C nuclear spins in diamond. Recent studies have shown that quantum memory technique or synchronized readout detection technique can further narrow down the spectral linewidth of NMR signal. In this short review paper, we overview basic concepts of nanoscale NMR using NV centers, and introduce further developments in high spectral resolution NV NMR studies.

Stability and Electronic Properties of the Adsorption of Molecular Hydrogen on Metal-containing Single-walled Carbon Nanotubes

  • Michael, Mananghaya
    • Journal of the Korean Chemical Society
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    • v.59 no.5
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    • pp.429-433
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    • 2015
  • The binding ability and hydrogen storage capacity of nitrogen doped carbon nanotube with divacancy (4NDCNxNT) that is decorated with transition metals was investigated based on density functional theory calculations. Results indicate that scandium shows an ideal reversible hydrogen binding capability with promising system-weight efficiency compared with other transition metals when functionalized with 4ND-CNxNT. The (Sc/4ND)10-CNxNT can store up to 50H2 molecules, corresponding to a maximum gravimetric density of 5.8 wt%. Detailed structural stability and electronic properties were reported as hydrogen molecules were absorbed. It takes about 0.16 eV/H2 to add one H2 molecule, which assures reversible storage of H2 molecules under ambient conditions.

Chirality of Drug Molecules and Biological Activity (의약품의 손대칭성과 생리 활성)

  • 권순경
    • Biomolecules & Therapeutics
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    • v.4 no.3
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    • pp.209-223
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    • 1996
  • Chirality is important in the context of biological activity because at a molecular level, asymmetry dominates biological process. While most pharmaceuticals of natural origin are single enantiomers, most of the synthesized chiral drugs are used in the form of racemic mixtures of two or more diastereomers. The enantiomers of a racemic drug generally differ in pharmacodynamic and pharmacokinetic properties as a consequence of stereoselective interaction with optically active molecular components of living organism. In pharmacokinetics and pharmacodynamics enantioselectivity plays an important role. The information on the sum of eutomer and distorter in a racemic drugs is very important in the estimation of therapeutic advantage and/or toxicity of racemates. The choice of preferentially developing a single enantiomer should be based on actual therapeutic advantages and especially improved safety.

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