Electroanalysis of unnatural base pair content in plasmid DNA generated in a semi-synthetic organism

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Authors

ŠPAČEK Jan FOJTA Miroslav

Year of publication 2020
Type Article in Periodical
Magazine / Source Electrochimica Acta
MU Faculty or unit

Central European Institute of Technology

Citation
Web https://www.sciencedirect.com/science/article/pii/S0013468620316911?via%3Dihub
Doi http://dx.doi.org/10.1016/j.electacta.2020.137298
Keywords Electrochemistry; Semi-synthetic organism; Unnatural base pair; Catalytic hydrogen evolution; Mercury electrode
Description While standard Watson-Crick base pairing normally involves hydrogen bonds that join size complemen-tary nitrogenous heterocycles, certain unnatural base pairs (UBP) can support replication, transcription, and translation, including these in semi-synthetic organisms (SSO), using size complementarity alone. In this paper we show that the UBPs can be analyzed electrochemically as components of riboor 2'-deoxyribonucleosides, corresponding (2'-deoxy)triphosphates, as well as incorporated into DNA. Here, performance of the electrochemical methods profits from the ability of the unnatural bases 5SICS and TPT3 adsorbed at the surface of a mercury electrode to catalyze hydrogen evolution in acidic media. This allows semi-quantitative detection of single UBP within plasmid DNA generated in a SSO. This electrochemical approach provides new way for direct, fast, and low cost UBP analysis and can supplement or replace currently used methods of UBP detection. (C) 2020 Elsevier Ltd. All rights reserved.
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