Národní úložiště šedé literatury Nalezeno 5 záznamů.  Hledání trvalo 0.00 vteřin. 
TAIL-LABELED OLIGONUCLEOTIDE PROBES FOR A DUAL ELECTROCHEMICAL MAGNETIC IMMUNOPRECIPITATION ASSAY OF DNA-PROTEIN BINDING
Hermanová, Monika ; Špaček, Jan ; Orság, Petr ; Fojta, Miroslav
A novel assay for detection of DNA-protein binding has been developed. Oligonucleotides bearing or lacking specific binding site of the p53 protein were tail-labeled by two different modified deoxynucleotide triphosphates using terminal deoxynucleotidyl transferase. Electrochemical detection enabled to discriminate between sequence-specific and non-specific p53-DNA binding in a competition assay.
Studies of Protein-DNA Interactions using Immunoprecipitation with DNA Probes Labelled with Electroactive Groups
Hermanová, Monika ; Špaček, Jan ; Pivoňková, Hana ; Fojta, Miroslav
Different electrochemically active species were tested for labelling of DNA probes and detection of DNA-protein binding. As the DNA probes, oligonucleotides bearing or lacking specific binding site of the p53 protein were chosen. They were tail-labelled either via chemical modification of single-stranded (ss) oligo(dT) DNA tails with an oxoosmium complex, or via enzymatic synthesis of the ss tail by terminal deoxynucleotidyl transferase and deoxynucleoside triphosphates modified with electrochemically reducible moieties. Electrochemical detection enabled to discriminate between sequence-specific and non-specific p53-DNA binding.
Novel Reactive Groups for Modification DNA by Oxoosmium Complexes
Havran, Luděk ; Vidláková, Pavlína ; Špaček, Jan ; Hermanová, Monika ; Fojta, Miroslav
DNA is naturally electroactive moleclue producing several intrinsie voltammetric signals. Some of them found appliction in electrochemical analysis of DNA interactions and damage. In particular types of applictaions is a suitable use DNA labelling by redox active tags to increase selectivity and sensitivity of analysis.One type of long-term used tags are complexes of osmium tetroxide with nittrogen ligunds (Os,L). These complexes preferentially react with pyrimidines in single stand DNA producing electractive adducts. primary reaction site for Os,L is C-C double bond in pyrimidine nucleobases. In this contribution will be presented results acquired with DNA containing chemically modified nuclebase.
Analysis of Products of Nontemplate Enzymatic Synthesis of DNA Oligonucleotides using Voltammetric Methods
Hermanová, Monika ; Havranová-Vidláková, Pavlína ; Ondráčková, Anna ; Fojta, Miroslav
Nontemplate DNA synthesis is a feature specific for a special type of DNA polymerase, terminal deoxynucleotidyl transferase (TdT). This enzyme is able. to randomly add nucleotides to a sintde-stranded DNA primer, provided the 3'OH end of the primer is accessible, Here we show that voltammetric analysis based on hanging mercury drop electrode (EPODE) and pyrolytic graphite electrode (PGE) enables to study this nontemplate DNA synthesis.
Electrochemical Study of Osmium Tetroxide Comptests Reactivity to DNA Bearing Butylacrylate
Havran, Luděk ; Havranová-Vidláková, Pavlína ; Špaček, Jan ; Vítová, Lada ; Hermanová, Monika ; Fojta, Miroslav
Coniplexes of osmium tetroside with nitrogen ligands (Os,L e.g.. with 2,2'-bipyridine (bpy) find application in redox labelling of DNA.. probing of DNA structure, and in studies of DNA interaction with other molecules Os,L preferentially react with pyrimidines in single strand DNA producing electroactive adducts Primary reaction site for Os,L is C=C double bond in pyrimidine nucleobases. In this contribution we introduce a new two-step technique of DNA modification with Os,bpy, consisting in enzymatic construdion or DNA beating butyl acrylate (BA) moieties attached to uracil or T-deaza adenine, followed by chemical modification of a reactive C=C double bond in BA residue.

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