Project Title

Genetic Characterization of a Klebsiella pneumoniae Secreted Anti-Microbial Protein

Author Names

Ethan BeckerFollow

Authors' Affiliations

Ethan Becker, Department of Health Sciences, College of Public Health, East Tennessee State University, Johnson City, TN

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

Classification of First Author

Undergraduate Student

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.

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Apr 6th, 9:15 AM Apr 6th, 9:30 AM

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.