Event Title

Characterizing E. coli promoters sensitive to subinhibitory concentration of antibiotics

Presenter Information

Anna Francis, Oberlin College

Location

Science Center Perlik Commons

Document Type

Poster

Start Date

5-13-2022 12:00 PM

End Date

5-13-2022 2:00 PM

Abstract

Antibiotic resistance arises due to the overexposure of antibiotics to bacteria, which can commonly occur at low concentrations of antibiotics over a long period of time. Our goal is to identify promoter regions in E. coli that are most sensitive to treatment of antibiotics at very low subinhibitory concentrations. This research will inform our development of whole-cell sensors that can detect subinhibitory concentrations of antibiotics in the environment. We are studying stress-responsive genes because we expect those to respond first to antibiotic treatment and initiate signaling cascade that aid in cell survival and fitness. Tetracycline is our antibiotic of interest because it is commonly found in human and animal waste and is difficult to eradicate from waste water. Our group has developed a dosing scheme for tetracycline below, at, and above its minimum inhibitory concentration (MIC), the concentration at which E. coli growth is inhibited. We have used techniques in RNA purification, polymerase chain reaction (PCR), and quantitative polymerase chain reaction (qPCR) to examine gene expression in response to low concentrations of tetracycline. Our findings indicate that treatment of half the MIC of tetracycline maximizes the expression of the genes baeR and spy, both of which are part of a periplasmic stress-responsive pathway. We found a number of time periods suitable for detecting increased gene expression, including 30 minutes and 120 minutes.

Keywords:

Antibiotics, Genes, Bacteria, Biochemistry

Major

Chemistry and Biochemistry

Project Mentor(s)

Lisa Ryno, Biochemistry

2022

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May 13th, 12:00 PM May 13th, 2:00 PM

Characterizing E. coli promoters sensitive to subinhibitory concentration of antibiotics

Science Center Perlik Commons

Antibiotic resistance arises due to the overexposure of antibiotics to bacteria, which can commonly occur at low concentrations of antibiotics over a long period of time. Our goal is to identify promoter regions in E. coli that are most sensitive to treatment of antibiotics at very low subinhibitory concentrations. This research will inform our development of whole-cell sensors that can detect subinhibitory concentrations of antibiotics in the environment. We are studying stress-responsive genes because we expect those to respond first to antibiotic treatment and initiate signaling cascade that aid in cell survival and fitness. Tetracycline is our antibiotic of interest because it is commonly found in human and animal waste and is difficult to eradicate from waste water. Our group has developed a dosing scheme for tetracycline below, at, and above its minimum inhibitory concentration (MIC), the concentration at which E. coli growth is inhibited. We have used techniques in RNA purification, polymerase chain reaction (PCR), and quantitative polymerase chain reaction (qPCR) to examine gene expression in response to low concentrations of tetracycline. Our findings indicate that treatment of half the MIC of tetracycline maximizes the expression of the genes baeR and spy, both of which are part of a periplasmic stress-responsive pathway. We found a number of time periods suitable for detecting increased gene expression, including 30 minutes and 120 minutes.