Development of an epigenetic tetracycline sensor system based on DNA methylation.
Anti-Bacterial Agents
/ analysis
AraC Transcription Factor
/ genetics
Biosensing Techniques
/ methods
DNA Methylation
Epigenomics
Escherichia coli
/ genetics
Escherichia coli Proteins
/ genetics
Flow Cytometry
/ methods
Operon
Plasmids
/ genetics
Repressor Proteins
/ genetics
Tetracycline
/ analysis
Journal
PloS one
ISSN: 1932-6203
Titre abrégé: PLoS One
Pays: United States
ID NLM: 101285081
Informations de publication
Date de publication:
2020
2020
Historique:
received:
09
03
2020
accepted:
19
04
2020
entrez:
8
5
2020
pubmed:
8
5
2020
medline:
4
8
2020
Statut:
epublish
Résumé
Bacterial live cell sensors are potentially powerful tools for the detection of environmental toxins. In this work, we have established and validated a flow cytometry readout for an existing bacterial arabinose sensor system with DNA methylation based memory function (Maier et al., 2017, Nat. Comm., 8:15336). Flow cytometry readout is convenient and enables a multiparameter analysis providing information about single-cell variability, which is beneficial for further development of sensor systems of this type in the future. We then designed a tetracycline sensor system, because of the importance of antibiotics pollution in the light of multi-resistant pathogens. To this end, a tetracycline trigger plasmid was constructed by replacing the araC repressor gene and the ara operator of the arabinose trigger plasmid with the tetR gene coding for the tetracycline repressor and the tet operon. After combination with the memory plasmid, the tetracycline sensor system was shown to be functional in E. coli allowing to detect and memorize the presence of tetracycline. Due to a positive feedback between the trigger and memory systems, the combined whole-cell biosensor showed a very high sensitivity for tetracycline with a detection threshold at 0.1 ng/ml tetracycline, which may be a general property of sensors of this type. Moreover, acute presence of tetracycline and past exposure can be detected by this sensor using the dual readout of two reporter fluorophores.
Identifiants
pubmed: 32379807
doi: 10.1371/journal.pone.0232701
pii: PONE-D-20-06852
pmc: PMC7205209
doi:
Substances chimiques
Anti-Bacterial Agents
0
AraC Transcription Factor
0
Escherichia coli Proteins
0
Repressor Proteins
0
tetracycline resistance-encoding transposon repressor protein
0
Tetracycline
F8VB5M810T
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e0232701Déclaration de conflit d'intérêts
The authors have declared that no competing interests exist.
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