Identification of the Targets of the Antimicrobial Peptide DASamP2 by Isolation of Pseudomonas Mutants with Increased Resistance

Advisor Information

Donald Rowen

Location

Dr. C.C. and Mabel L. Criss Library

Presentation Type

Poster

Start Date

6-3-2015 11:00 AM

End Date

6-3-2015 12:30 PM

Abstract

The emergence of bacteria that have developed resistance to current antibiotics is an issue of great concern because we may no longer be able to treat some bacterial infections. Antimicrobial peptides (AMP) have shown promise as a source of new antimicrobial agents. Antimicrobial peptides are short proteins with a wide range of antimicrobial activity against bacteria, fungi, and viruses; they can vary from 12-50 amino acids in length. Dr. Guangshun Wang at UNMC has developed a promising new AMP (DASamP2) that is effective against Psedomonas aeruginosa as well as many other types of bacterium. To further investigate DASamp2, we are seeking to determine its mechanism of action against bacteria.cells. To do this, we sought to isolate mutants of the bacterium P. aeruginosa that show increased resistance to the AMP. These mutants are expected to have a mutation in a gene that encodes a target of the AMP. We made mutants by causing a transposon to jump randomly into the P. aeruginosa genome. I helped to screen 1,500 transposon mutant cells for increased resistance. Mutants that showed increase resistance in an initial screen were retested. I ultimately helped to isolate four mutants that showed increased resistance after three rounds of retesting. Determining the location of the transposon in these four mutants should help to identify the mechanism of action of DASamp2.

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Mar 6th, 11:00 AM Mar 6th, 12:30 PM

Identification of the Targets of the Antimicrobial Peptide DASamP2 by Isolation of Pseudomonas Mutants with Increased Resistance

Dr. C.C. and Mabel L. Criss Library

The emergence of bacteria that have developed resistance to current antibiotics is an issue of great concern because we may no longer be able to treat some bacterial infections. Antimicrobial peptides (AMP) have shown promise as a source of new antimicrobial agents. Antimicrobial peptides are short proteins with a wide range of antimicrobial activity against bacteria, fungi, and viruses; they can vary from 12-50 amino acids in length. Dr. Guangshun Wang at UNMC has developed a promising new AMP (DASamP2) that is effective against Psedomonas aeruginosa as well as many other types of bacterium. To further investigate DASamp2, we are seeking to determine its mechanism of action against bacteria.cells. To do this, we sought to isolate mutants of the bacterium P. aeruginosa that show increased resistance to the AMP. These mutants are expected to have a mutation in a gene that encodes a target of the AMP. We made mutants by causing a transposon to jump randomly into the P. aeruginosa genome. I helped to screen 1,500 transposon mutant cells for increased resistance. Mutants that showed increase resistance in an initial screen were retested. I ultimately helped to isolate four mutants that showed increased resistance after three rounds of retesting. Determining the location of the transposon in these four mutants should help to identify the mechanism of action of DASamp2.