Antibiotic resistance is a significant public health threat; even low concentrations of antibiotics still contribute to high levels of resistance. We are interested in constructing fluorescent sensors that detect low concentrations of antibiotics in complex solutions. We have used plasmids with a selective toxin-antitoxin system and green fluorescent protein (GFP). Using standard molecular biology techniques, we deleted the neomycin/kanamycin resistance cassette from this plasmid. We are currently working to insert different promoter regions corresponding to stress-responsive signaling pathways. To identify sensitive promoter regions of interest, we are using quantitative polymerase chain reaction to monitor transcript levels of several genes in stress-responsive signaling pathways after <i>E. coli </i>are exposed to various low antibiotic concentrations. Preliminary results focused on studying the aminoglycoside gentamicin by observing the activity of several transcriptional pathways involved in the periplasmic stress response. We monitored transcripts of genes in the BaeR, CpxR, and RpoE stress-responsive pathways. Future work includes adding different transcription factor binding sites as the promoter region ahead of the GFP in our exogenous plasmids and testing the sensitivity of our sensors by monitoring GFP fluorescence after treatment with different classes and concentrations of antibiotics.

Development of whole-cell sensors for detection of antibiotics at low concentrations

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