Vertaisarvioitu alkuperäisartikkeli tai data-artikkeli tieteellisessä aikakauslehdessä (A1)
A Plasmid-Based Fluorescence Reporter System for Monitoring Oxidative Damage in E. coli
Julkaisun tekijät: Dandapani Hariharan, Kankaanpää Pasi, Jones Patrik R, Kallio Pauli
Kustantaja: MDPI
Julkaisuvuosi: 2022
Journal: Sensors
Tietokannassa oleva lehden nimi: SENSORS
Lehden akronyymi: SENSORS-BASEL
Volyymi: 22
Julkaisunumero: 17
Sivujen määrä: 15
DOI: http://dx.doi.org/10.3390/s22176334
Verkko-osoite: https://www.mdpi.com/1424-8220/22/17/6334
Rinnakkaistallenteen osoite: https://research.utu.fi/converis/portal/detail/Publication/176534979
Quantitating intracellular oxidative damage caused by reactive oxygen species (ROS) is of interest in many fields of biological research. The current systems primarily rely on supplemented oxygen-sensitive substrates that penetrate the target cells, and react with ROS to produce signals that can be monitored with spectroscopic or imaging techniques. The objective here was to design a new non-invasive analytical strategy for measuring ROS-induced damage inside living cells by taking advantage of the native redox sensor system of E. coli. The developed plasmid-based sensor relies on an oxygen-sensitive transcriptional repressor IscR that controls the expression of a fluorescent marker in vivo. The system was shown to quantitatively respond to oxidative stress induced by supplemented H2O2 and lowered cultivation temperatures. Comparative analysis with fluorescence microscopy further demonstrated that the specificity of the reporter system was equivalent to the commercial chemical probe (CellROX). The strategy introduced here is not dependent on chemical probes, but instead uses a fluorescent expression system to detect enzyme-level oxidative damage in microbial cells. This provides a cheap and simple means for analysing enzyme-level oxidative damage in a biological context in E. coli.
Ladattava julkaisu This is an electronic reprint of the original article. |  |
