Chromogenic Escherichia coli reporter strain for screening DNA damaging agents.
Josué Daniel Mora-GarduñoJessica Tamayo-NuñezFelipe Padilla-VacaFátima Berenice Ramírez-MontielÁngeles Rangel-SerranoFernando Santos-EscobarFélix Gutiérrez-CoronaItzel Páramo-PérezFernando Anaya-VelázquezRodolfo García-ContrerasNaurú Idalia Vargas-MayaBernardo FrancoPublished in: AMB Express (2022)
The presence of pollutants in soil and water has given rise to diverse analytical and biological approaches to detect and measure contaminants in the environment. Using bacterial cells as reporter strains represents an advantage for detecting pollutants present in soil or water samples. Here, an Escherichia coli reporter strain expressing a chromoprotein capable of interacting with soil or water samples and responding to DNA damaging compounds is validated. The reporter strain generates a qualitative signal and is based on the expression of the coral chromoprotein AmilCP under the control of the recA promoter. This strain can be used simply by applying soil or water samples directly and rendering activation upon DNA damage. This reporter strain responds to agents that damage DNA (with an apparent detection limit of 1 µg of mitomycin C) without observable response to membrane integrity damage, protein folding or oxidative stress generating agents, in the latter case, DNA damage was observed. The developed reporter strain reported here is effective for the detection of DNA damaging agents present in soils samples. In a proof-of-concept analysis using soil containing chromium, showing activation at 15.56 mg/L of Cr(VI) present in soil and leached samples and is consistent with Cr(III) toxicity at high concentrations (130 µg). Our findings suggest that chromogenic reporter strains can be applied for simple screening, thus reducing the number of samples requiring analytical techniques.
Keyphrases
- oxidative stress
- escherichia coli
- dna damage
- crispr cas
- circulating tumor
- single molecule
- induced apoptosis
- cell free
- heavy metals
- dna repair
- dna methylation
- gene expression
- ischemia reperfusion injury
- diabetic rats
- staphylococcus aureus
- multidrug resistant
- cell death
- pseudomonas aeruginosa
- binding protein
- computed tomography
- protein kinase
- endoplasmic reticulum stress
- cystic fibrosis
- liquid chromatography