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Carbon Electrodes for Enzyme-Based Voltammetric Detection of DNA Hybridization: Detection of DNA Sequence from the Aflatoxinogenic Fungi
Authors: Šnévajsová Petra | Tisoň Lukáš | Vytřasová Jarmila
Year: 2009
Type of publication: ostatní - článek ve sborníku
Name of source: Abstracts - XXth International Symposium on Bioelectrochemistry and Bioenergetics 2009
Publisher name: Romanian Society of Pure and Applied Biophysic
Place: Bucharest
Page from-to: 152-153
Titles:
Language Name Abstract Keywords
cze Uhlíkové elektrody provoltametrickou detekci hybridizace DNA: Detekce specifickéhoúseku DNA aflatoxinogenních plísní Cílem práce je zavést metodiku pro elektrochemickou detekci vybraného úseku DNA kódujícího biochemickou cestu syntézy aflatoxinu B1 u některých kmenů plísní rodu Aspergillus. Princip metody se zakládá na aktivitě enzymu alkalické fosfatázy (ALP). Cílová DNA je nejdříve imobilizována na povrch elektrody, poté hybridizuje se sondou značenou biotinem. Na biotin je navázán konjugát streptavidinu s ALP. Alkalická fosfatáza katalyzuje přeměnu elektroinaktivního substrátu 1-naftylfosfátu na elektroaktivní produkt 1-naftol, jehož signál je detekován voltametricky. Aspergillus; carbon paste; DNA
eng Carbon Electrodes for Enzyme-Based Voltammetric Detection of DNA Hybridization: Detection of DNA Sequence from the Aflatoxinogenic Fungi In the present study we are exploring possibilities for electrochemical detection of a regulatory gene encoding for a biochemical pathway of aflatoxins. In our contribution, target DNA was immobilized by adsorption onto a surface of the carbon electrode. In order to avoid non-specific adsorption onto electrode surface, unoccupied surface was blocked by bovine serum albumin and hybridization between the target DNA and biotin-DNA probe followed. Then the conjugate of streptavidin with alkaline phosphatase was added and bound to the biotin labelled probe. The electrode was placed into voltammetric cell containing buffer and 1-naphthyl phosphate. The 1- naphthol product of enzymatic reaction quantified voltammetric technique. Aspergillus; carbon paste; DNA

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