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Electron Microscopic Visualization of Complementary Labeled DNA With Platinum-Containing Guanine Derivative
Saim Emin, Svetlin TOSHEV, Alexandre Loukanov, Chavdar Filipov, Polina Mladenova, 2016, izvirni znanstveni članek

Opis: The object of the present report is to provide a method for a visualization of DNA in TEM by complementary labeling of cytosine with guanine derivative, which contains platinum as contrast-enhanced heavy element. The stretched single-chain DNA was obtained by modifying double-stranded DNA. The labeling method comprises the following steps: (i) stretching and adsorption of DNA on the support film of an electron microscope grid (the hydrophobic carbon film holding negative charged DNA); (ii) complementary labeling of the cytosine bases from the stretched single-stranded DNA pieces on the support film with platinum containing guanine derivative to form base-specific hydrogen bond; and (iii) producing a magnified image of the base-specific labeled DNA. Stretched single-stranded DNA on a support film is obtained by a rapid elongation of DNA pieces on the surface between air and aqueous buffer solution. The attached platinum-containing guanine derivative serves as a high-dense marker and it can be discriminated from the surrounding background of support carbon film and visualized by use of conventional TEM observation at 100 kV accelerated voltage. This method allows examination of specific nucleic macromolecules through atom-by-atom analysis and it is promising way toward future DNA-sequencing or molecular diagnostics of nucleic acids by electron microscopic observation.
Najdeno v: osebi
Ključne besede: DNA stretching and visualization, contrast-enhanced Pt-containing guanine, DNA sequences
Objavljeno: 02.02.2017; Ogledov: 2192; Prenosov: 0
.pdf Polno besedilo (298,70 KB)

Biotinylated Vanadium and Chromium Sulfide Nanoparticles as Probes for Colocalization of Membrane Proteins
Saim Emin, Alexandre Loukanov, 2016, izvirni znanstveni članek

Opis: ABSTRACT We report the microemulsion synthesis of vanadium and chromium sulfide nanoparticles (NPs) and their biological application as nanoprobes for colocalization of membrane proteins. Spherical V2S3 and Cr2S3 NPs were prepared in reverse microemulsion droplets, as nanoreactors, obtained by the surfactant sodium bis(2-ethylhexyl) sulfosuccinate (AOT) in nonpolar organic phase (heptane). Electron microscopic data indicated that the size distribution of the nanoparticles was uniform with an average diameter between 3 4 5 nm. The prepared hydrophobic nanocrystals were transferred in aqueous phase by surface cap exchange of AOT with biotindihydrolipoic ligands. This substitution allows the nanoparticles solubility in aqueous solutions and confer their bioactivity. In addition, we report the conjugation procedure between a-Lipoic acid (LA) and biotin (abbreviated as biotin-LA). The biotin-LA structure was characterized by 1D and 2D NMR spectroscopy. The biotinylated vanadium and chromium sulfide nanoparticles were tested as probes for colocalization of glutamate receptors on sodium-dodecyl-sulfate-digested replica prepared from rat hippocampus. The method suggests their high labeling efficiency for study of membrane biological macromolecules.
Najdeno v: osebi
Ključne besede: biotin-dihydrolipoic conjugate, vanadium and chromium sulfide nanoparticles, membrane proteins
Objavljeno: 02.02.2017; Ogledov: 2286; Prenosov: 0
.pdf Polno besedilo (581,45 KB)

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