Presentation Title

Identification of Mechanism of Action of DASamP2 against Pseudomonas aeruginosa by Isolation of Mutants

Advisor Information

Donald Rowen

Location

UNO Criss Library, Room 231

Presentation Type

Oral Presentation

Start Date

6-3-2015 2:30 PM

End Date

6-3-2015 2:45 PM

Abstract

Human bacterial pathogens are evolving resistance to antibiotics currently used for therapy. There is a need to develop new strategies to combat resistant microbes such as Staphylococcus aureus and Pseudomonas aeruginosa which exhibit multi-drug resistance. Antimicrobial peptides are diverse groups of molecules that possess antimicrobial properties (AMPs) that are being studied for use as antibiotics. They are naturally produced by animals as a defense mechanism against microorganisms. The mechanism of action of antimicrobial peptides is not well known. Most are thought to disrupt cell membranes, but some may inhibit the activity of a cytosolic biosynthetic enzyme. Recently a new antimicrobial peptide (DASamp2) designed by Dr. Wang (University of Nebraska Medical Center) was observed to be effective against both S. aureus and P. aeruginosa and thereby holds promise as a potential new antibiotic. The goal of our project is to determine the mechanism of action and target of the DASamp2 by isolating mutants of P. aeruginosa with altered sensitivity to the DASamp2 peptide. We are performing transposon mutagenesis of the P. aerugionosa strain PAO1 and screening for increased and decreased resistance to DASamp2. So far, we have screened 3600 mutants for increased resistance and isolated ten resistant mutants. We have begun to identify the genes mutated in the mutants isolated by using inverse PCR. Characterization of the genes mutated will help in elucidating the target and mechanism action of the DASamp2 peptide.

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COinS
 
Mar 6th, 2:30 PM Mar 6th, 2:45 PM

Identification of Mechanism of Action of DASamP2 against Pseudomonas aeruginosa by Isolation of Mutants

UNO Criss Library, Room 231

Human bacterial pathogens are evolving resistance to antibiotics currently used for therapy. There is a need to develop new strategies to combat resistant microbes such as Staphylococcus aureus and Pseudomonas aeruginosa which exhibit multi-drug resistance. Antimicrobial peptides are diverse groups of molecules that possess antimicrobial properties (AMPs) that are being studied for use as antibiotics. They are naturally produced by animals as a defense mechanism against microorganisms. The mechanism of action of antimicrobial peptides is not well known. Most are thought to disrupt cell membranes, but some may inhibit the activity of a cytosolic biosynthetic enzyme. Recently a new antimicrobial peptide (DASamp2) designed by Dr. Wang (University of Nebraska Medical Center) was observed to be effective against both S. aureus and P. aeruginosa and thereby holds promise as a potential new antibiotic. The goal of our project is to determine the mechanism of action and target of the DASamp2 by isolating mutants of P. aeruginosa with altered sensitivity to the DASamp2 peptide. We are performing transposon mutagenesis of the P. aerugionosa strain PAO1 and screening for increased and decreased resistance to DASamp2. So far, we have screened 3600 mutants for increased resistance and isolated ten resistant mutants. We have begun to identify the genes mutated in the mutants isolated by using inverse PCR. Characterization of the genes mutated will help in elucidating the target and mechanism action of the DASamp2 peptide.