Related Experiment Videos
Multiple antibiotic susceptibility associated with inactivation of the prc gene
A Seoane1, A Sabbaj, L M McMurry
1Department of Molecular Biology, Tufts University School of Medicine, Boston, Massachusetts.
Abstract:
A Tn5 insertion which led to increased susceptibility to multiple drugs, including tetracycline, chloramphenicol, nalidixic acid, erythromycin, spectinomycin, norfloxacin, and novobiocin, was identified in Escherichia coli. Cloning and sequence studies showed that the insertion was in the previously identified prc gene at min 40.4. The prc product is known to function as a protease linked to processing of penicillin-binding protein 3 and lambda repressor and when absent to allow some leakage of periplasmic constituents. Complementation studies with the prc gene on plasmids showed complete recovery of parental levels of susceptibility to all drugs except chloramphenicol, with which only partial reversion to wild-type levels was observed.
Insights
A genetic mutation in Escherichia coli
Area of Science:
- Microbiology
- Molecular Biology
- Genetics
Background:
- The prc gene in Escherichia coli encodes a protease involved in processing penicillin-binding protein 3 and lambda repressor.
- Loss of prc function can lead to leakage of periplasmic constituents.
- Antibiotic resistance is a significant challenge in microbial infections.
Purpose of the Study:
- To investigate the genetic basis of increased multidrug susceptibility in Escherichia coli.
- To identify the specific gene mutation responsible for altered drug resistance profiles.
- To understand the role of the prc gene in antibiotic susceptibility.
Main Methods:
- Utilized Tn5 mutagenesis to generate drug-susceptible mutants in Escherichia coli.
- Performed cloning and DNA sequencing to identify the insertion site.
- Conducted complementations studies using plasmids carrying the prc gene.
Main Results:
- A Tn5 insertion within the prc gene at min 40.4 resulted in increased susceptibility to multiple antibiotics.
- Complementation with a functional prc gene fully restored susceptibility to most tested drugs.
- Partial restoration of wild-type chloramphenicol susceptibility was observed after complementation.
Conclusions:
- The prc gene plays a crucial role in maintaining wild-type levels of resistance to several antibiotics in Escherichia coli.
- Disruption of the prc gene significantly impacts the bacterium's susceptibility profile.
- Targeting the prc gene or its product could be a potential strategy to combat multidrug-resistant bacteria.