• Title/Summary/Keyword: Species affinity

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In Vitro Selection of RNA Aptamer Specific to Salmonella Typhimurium

  • Han, Seung Ryul;Lee, Seong-Wook
    • Journal of Microbiology and Biotechnology
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    • v.23 no.6
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    • pp.878-884
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    • 2013
  • Salmonella is a major foodborne pathogen that causes a variety of human diseases. Development of ligands directly and specifically binding to the Salmonella will be crucial for the rapid detection of, and thus for efficient protection from, the virulent bacteria. In this study, we identified a RNA aptamer-based ligand that can specifically recognize Salmonella Typhimurium through SELEX technology. To this end, we isolated and characterized an RNase-resistant RNA aptamer that bound to the OmpC protein of Salmonella Typhimurium with high specificity and affinity ($K_d$ ~ 20 nM). Of note, the selected aptamer was found to specifically bind to Salmonella Typhimurium, but neither to Gram-positive bacteria (Staphylococcus aureus) nor to other Gram-negative bacteria (Escherichia coli O157:H7). This was evinced by aptamer-immobilized ELISA and aptamer-linked precipitation experiments. This Salmonella species-specific aptamer could be useful as a diagnostic ligand against pathogen-caused foodborne sickness.

Photocatalytic Epoxidation of Olefins Using Molecular O2 by TiO2 Incorporated in Hydrophobic Y Zeolite

  • Kuwahara, Yasutaka;Magatani, Yasuhiro;Yamashita, Hiromi
    • Rapid Communication in Photoscience
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    • v.4 no.1
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    • pp.19-21
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    • 2015
  • Zeolite is an ideal host material for encapsulating nano-size metal catalyst species because of its defined microporous structure, prominent adsorption/condensation properties, high surface area, chemical/thermal stability, and transparency to light. In this study, $TiO_2$ photocatalyst was incorporated in highly hydrophobic Y zeolite and its photocatalytic activity was examined in the photocatalytic oxidation of olefins under UV-light irradiation using molecular oxygen as an oxygen source. $TiO_2$ nanoparticles incorporated in hydrophobic Y zeolite exhibited a markedly enhanced photocatalytic activity compared with bare $TiO_2$ owing to its excellent affinity toward organic moieties, which facilitates the mass transfer of organic substrates and allows them to efficiently access to the neighboring active $TiO_2$ surface.

Phase Behavior of Reversibly Associating Star Copolymer-like Polymer Blends

  • June Huh;Kim, Seung-Hyun;Jo, Won-Ho
    • Macromolecular Research
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    • v.10 no.1
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    • pp.18-23
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    • 2002
  • We theoretically consider blends of two monodisperse one-end-functionalized homopolymers (denoted by A and B) capable of forming clusters between functional groups (stickers) using weak segregation theory. In this model system resulting molecular architectures via clustering resemble star copolymers having many A- and B-arms. Minimizing the total free energy with respect the cluster distribution, the equilibrium distribution of clusters is obtained and used for RPA (Random Phase Approximation) equations as input. For the case that polymers are functionalized by only one kind of sticker, the phase diagrams show that the associations promote the macrophase separation. When there is strong affinity between stickers belonging to the different polymer species, on the other hand, the phase diagram show a suppression of the macrophase separation at the range of high temperature regime, as well as the phase coexistence between a disordered and a mesoscopic phase at the relatively lower temperatures.

Label-free Detection of Biomolecular Specific Interaction by Optical Biosensors (광 바이오센서를 이용한 비표지 생계물질들의 특이 상호작용력의 측정)

  • 김의락;최정우
    • KSBB Journal
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    • v.17 no.1
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    • pp.1-13
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    • 2002
  • Label-free optical methods for the monitoring of interactions between biological molecules have become increasingly popular within the last decade. A rising number of publications have demonstrated the benefits of direct biomolecular interaction analysis(BIA) for biology and biochemistry, such as antigen-antibody Interactions, receptor-ligand interactions, protein-DNA, DNA- intercalator, and DNA-DNA interactions. This article gives an overview of the historical development, principle and application of label-free optical biosensor to examine the functional characteristics of biospecific interaction, such as kinetics, affinity, and binding position of biomolecular between an immobilized species at the transducer surface and its dissolved binding partner.

Effective Uranyl Binding by a Dihydroxyazobenzene Derivative. Ionization of Uranium-Bound Water

  • 이관표;장보빈;서정훈
    • Bulletin of the Korean Chemical Society
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    • v.17 no.9
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    • pp.814-819
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    • 1996
  • In search of simple host molecules for uranyl ion which form 1: 1-type complexes with high formation constants that can be used either in extraction of uranium from seawater or in catalysis of biologically important organic reactions, the uranophile activities of dihydroxyazobenzene derivative 1 were studied. Uranyl ion and 1 form a 1: 1-type complex with a very large formation constant. The formation constant was measured at pH 7-11.6 by competition experiments with carbonate ion. From the resulting pH dependence, ionization constants of the two aquo ligands coordinated to the uranium of the uranyl complex of 1 were calculated. The ionization constants were also measured by potentiometric titration of the uranyl complex of 1. Based on these results, the pKa values of the two aquo ligands were estimated as 7.1 and 11.0, respectively. At pH 7.5-9.5, therefore, the complex exists mostly as monohydroxo species. Under the conditions of seawater, 1 possesses greater affinity toward uranyl ion compared with other uranophiles such as carbonate ion, calixarene derivatives, or a macrocyclic octacarboxylate. In addition, complexation of 1 with uranyl ion is much faster than that of the calixarene or octacarboxylate uranophiles.

Purification and Characterization of Repressor of Temperate S. aureus Phage Φ11

  • Das, Malabika;Ganguly, Tridib;Chattoraj, Partho;Chanda, Palas Kumar;Bandhu, Amitava;Lee, Chia Yen;Sau, Subrata
    • BMB Reports
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    • v.40 no.5
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    • pp.740-748
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    • 2007
  • To gain insight into the structure and function of repressor proteins of bacteriophages of gram-positive bacteria, repressor of temperate Staphylococcus aureus phage ${\phi}11$ was undertaken as a model system here and purified as an N-terminal histidine-tagged variant (His-CI) by affinity chromatography. A ~19 kDa protein copurified with intact His-CI (~ 30 kDa) at low level was resulted most possibly due to partial cleavage at its Ala-Gly site. At ~10 nM and higher concentrations, His-CI forms significant amount of dimers in solution. There are two repressor binding sites in ${\phi}11$ cI-cro intergenic region and binding to two sites occurs possibly by a cooperative manner. Two sites dissected by HincII digestion were designated operators $O_L$ and $O_R$, respectively. Equilibrium binding studies indicate that His-CI binds to $O_R$ with a little more strongly than $O_L$ and binding species is probably dimeric in nature. Interestingly His-CI binding affinity reduces drastically at elevated temperatures ($32-42^{\circ}C$). Both $O_L$ and $O_R$ harbor a nearly identical inverted repeat and studies show that ${\phi}11$ repressor binds to each repeat efficiently. Additional analyses indicate that ${\phi}11$ repressor, like $\lambda$ repressor, harbors an N-terminal domain and a C-terminal domain which are separated by a hinge region. Secondary structure of ${\phi}11$ CI even nearly resembles to that of $\lambda$ phage repressor though they differ at sequence level. The putative N-terminal HTH (helix-turn-helix) motif of ${\phi}11$ repressor belongs to the HTH -XRE-family of proteins and shows significant identity to the HTH motifs of some proteins of evolutionary distant organisms but not to HTH motifs of most S. aureus phage repressors.

Construction of Recombinant DNA for Purification of the Gag-Pro Transframe Protein of Human T-cell Leukemia Virus Type I (HTLV-I) (Human T-cell Leukemia Virus Type I (HTLV-I) 의 Gag-Pro Transframe 단백질 정제를 위한 재조합 DNA 의 제작)

  • 남석현
    • Korean Journal of Microbiology
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    • v.30 no.6
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    • pp.466-471
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    • 1992
  • To determine the site at which -1 ribosomal frameshifting occurs within the gag-pro overlap of HTL V-I. DNA fragment corresponding to a portion of the gene overlap was cloned into a SP6 vector. The resultant plasmid harbors the hybrid gene consisting of a synthetic gene encoding 5 amino acids derived from chick prelysozyme including the initiator methionine plus 141 nucleotides of gag-pro overlapping region followed by Staphylococcus aurcus protein A gene fragment. In vitro transcription by SP6 RNA polymerase with this DNA template made an abundant amount of single species mRNA. Cell-free translation programmed with the RNA transcribed in vitro yielded a polypeptide of 21 kDal in size. which could be purified into homogeneity by IgG-Sepharose affinity chromatography. In vitro system described in this study must be useful for rapid purification and sequencing of the Gag-Pro transframe protein. allowing to determine the exact frameshift site on mRNA and to identify the tRNA involved in frameshifting event for the expression of pro gene.

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Amperometric Enzyme Electrode for the Determination of $NH_4^+$ ($NH_4^+$ 정량을 위한 Amperometric Enzyme Electrode)

  • Moo Lyong Seo;Jae Sang Kim;Shim Sung Lee;Zun Ung Bae;Heung Lark Lee;Tae Myung Park
    • Journal of the Korean Chemical Society
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    • v.37 no.11
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    • pp.937-942
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    • 1993
  • Enzyme electrodes for amperometric measurement of ammonia was prepared by immobilization of L-glutamate dehydrogenase on an Immobilon-AV Affinity membrane and attachment to a glassy carbon electrode. Reduced nicotinamide adenine dinucleotide (NADH) was used as the electroactive species. The electrochemical oxidation of NADH was monitored at +1.0 volt vs. Ag/AgCl. Response was linear from $4.0\;{\times}\;10^{-5}\;to\;4.0\;{\times}\;10^{-4}$ M. The detection limit was 2.0 ${\times}\;10^{-6}$ M. Response time, the optimum pH and life time of enzyme immobilized membrane were 2 min, pH 7.3∼7.6 (Dulbecco's buffer solution) and about 25 days respectively. When the enzyme electrode was applied to the $NH_4^+$ determination with amperometric method, other physiological materials had no interference.

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Removal of Arsenite and Arsenate by a Sand Coated with Colloidal Hematite Particl (나노 크기 적철석 입자 피복 모래를 이용한 비소 3가와 비소 5가의 제거)

  • 고일원;이상우;김주용;김경웅;이철효
    • Journal of Soil and Groundwater Environment
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    • v.9 no.1
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    • pp.63-69
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    • 2004
  • Hematite-coated sand was examined for the application of the PRB (permeable reactive barrier) to the arsenic-contaminated subsurface in the metal mining areas. The removal efficiency of As in a batch and a flow system was investigated through the adsorption isotherm, removal kinetics and column experiments. Hematite-coated sand followed a linear adsorption isotherm with high adsorption capacity at low level concentrations of As (<1.0 mg/L). In the column experiments, high content of hematite-coated sand enhanced the removal efficiency, but the amount of the As removal decreased due to the higher affinity of As (V) than As (III) and reduced adsorption kinetics in the flow system. Therefore. the amount of hematite-coated sand, the adsorption affinity of As species and removal kinetics determined the removal efficiency of As in a flow system.

How to Design Membrane Chromatography for Bioseparations: A Short Review (바이오분야 분리용 막크로마토그래피 설계 방안)

  • Park, Inho;Yoo, Seung Yeon;Park, Ho Bum
    • Membrane Journal
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    • v.31 no.2
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    • pp.145-152
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    • 2021
  • While there are increasing demands on biomolecules separation, resin chromatography lacks in terms of throughput and membrane chromatography is an alternative with high binding capacity and enhanced mass transfer properties. Unlike typical membrane processing, where the performance can only be empirically assessed, understanding how mechanisms work in membrane chromatography is decisive to design biospecific processing. This short review covers three separation mechanisms, including affinity interaction modes for selectively capturing bulk molecules using biospecific sites, ion exchange modes for binding biomolecules using net charges and hydrophobic interaction modes for binding targeted, hydrophobic species. The parameters in designing membrane chromatography that should be considered operation-wise or material-wise, are also further detailed in this paper.