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


Tiivistelmä
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.

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Last updated on 2022-06-10 at 10:46