Genetic Characterization of a Klebsiella pneumoniae Secreted Anti-Microbial Protein
Location
Culp Room 217
Start Date
4-6-2022 9:15 AM
End Date
4-6-2022 9:30 AM
Faculty Sponsor’s Department
Health Sciences
Name of Project's Faculty Sponsor
Sean Fox
Additional Sponsors
Dr. Ranjan Chakraborty Dr. Erik Peterson
Competition Type
Non-Competitive
Type
Boland Symposium
Project's Category
Microbiology, Molecular Biology
Abstract or Artist's Statement
Antimicrobial-resistant (AMR) bacteria are a major source of concern in modern-day nosocomial settings, leading to possible further drug resistance or spread to those who cannot fight off the infection. Previous work from our laboratory has shown that Klebsiella pneumoniae (KP) secretes an antimicrobial protein that has been shown to inhibit the growth of many species of bacteria that contain AMR properties in the Enterobacteriaceae family, a major contributor to nosocomial AMR. Klebsiella spent media is able to inhibit the growth of Citrobacter freundii (CF), Enterobacter aerogenes (EA), and Enterobacter cloacae (ECL) through an anti-microbial protein (AMP). This AMP has been shown to reduce the density and growth of CF, EA, and ECL in both biofilm and planktonic forms. To determine the genetic elements involved in AMP production, we introduced a transposon (Tn5) into the genome of Klebsiella to provide resistant selection and to create a mutant knockout to find the exact location of the gene. Upon transposon mutagenesis, the resulting genome was electroporated into Rec- E. coli. The E. coli was now able to produce the antimicrobial protein, with the zones of inhibition for CF, EA, and ECL. Upon confirmation that the plasmid mediates the AMP, the plasmid was sent for sequencing to further characterize the gene responsible for coding the AMP. This newly identified AMP may prove to be a valuable treatment for AMR bacteria once characterized.
Genetic Characterization of a Klebsiella pneumoniae Secreted Anti-Microbial Protein
Culp Room 217
Antimicrobial-resistant (AMR) bacteria are a major source of concern in modern-day nosocomial settings, leading to possible further drug resistance or spread to those who cannot fight off the infection. Previous work from our laboratory has shown that Klebsiella pneumoniae (KP) secretes an antimicrobial protein that has been shown to inhibit the growth of many species of bacteria that contain AMR properties in the Enterobacteriaceae family, a major contributor to nosocomial AMR. Klebsiella spent media is able to inhibit the growth of Citrobacter freundii (CF), Enterobacter aerogenes (EA), and Enterobacter cloacae (ECL) through an anti-microbial protein (AMP). This AMP has been shown to reduce the density and growth of CF, EA, and ECL in both biofilm and planktonic forms. To determine the genetic elements involved in AMP production, we introduced a transposon (Tn5) into the genome of Klebsiella to provide resistant selection and to create a mutant knockout to find the exact location of the gene. Upon transposon mutagenesis, the resulting genome was electroporated into Rec- E. coli. The E. coli was now able to produce the antimicrobial protein, with the zones of inhibition for CF, EA, and ECL. Upon confirmation that the plasmid mediates the AMP, the plasmid was sent for sequencing to further characterize the gene responsible for coding the AMP. This newly identified AMP may prove to be a valuable treatment for AMR bacteria once characterized.