Un-Masking Hidden Phenotypes for PA14 IVYp1 and IVYp2 Knockout Strains by Observation of Tetracycline Sensitivity and Restoration of Resistance
Disciplines
Medicine and Health Sciences
Abstract (300 words maximum)
Pseudomonas aeruginosa (PA) is a gram-negative bacteria that is known for its adaptive antimicrobial resistance. Inhibitors of vertebrate lysozyme proteins IVY’s are periplasmic proteins that are involved in the resistome of PA. They are deemed responsible for this resistivity, based on previous findings that demonstrate a tetracycline-sensitive phenotype for IVYp1 knockout under growth conditions in parallel with cystic fibrosis lung environment. However, this resistivity has not been tested using standard laboratory conditions. The aim of this CSM mentor protégé funded project is to implement standard microbiological characterization such as Kirby-Bauer Susceptibility Test to provide qualitative results en par with Minimum Inhibitory Concentration assays to indicate tetracycline sensitivity of the IVYp1 knockout. Assessing these results from Kirby Bauer test, the minimum inhibitory concentrations for the knockout strains will be determined to obtain the minimum amount of antibiotic needed to limit the growth for wild type, IVYp1-, and IVYp2- strains of PA14. Ongoing studies to use complementation of IVYp1 inactivation loop by exogenously added peptide are being used to further indicate the sustainability of IVYp1. IVYp1-/- strain will be examined under the addition of a cyclic peptide consisting of six amino acids followed by MIC assays performed to evaluate if the phenotype TetR is restored through the exogenously added peptide advocating for the necessity and sufficiency of IVYp1 for maintaining the integrity of peptidoglycan cell wall, thus making it a potential drug target.
Academic department under which the project should be listed
CSM - Chemistry and Biochemistry
Primary Investigator (PI) Name
Thomas C. Leeper
Un-Masking Hidden Phenotypes for PA14 IVYp1 and IVYp2 Knockout Strains by Observation of Tetracycline Sensitivity and Restoration of Resistance
Pseudomonas aeruginosa (PA) is a gram-negative bacteria that is known for its adaptive antimicrobial resistance. Inhibitors of vertebrate lysozyme proteins IVY’s are periplasmic proteins that are involved in the resistome of PA. They are deemed responsible for this resistivity, based on previous findings that demonstrate a tetracycline-sensitive phenotype for IVYp1 knockout under growth conditions in parallel with cystic fibrosis lung environment. However, this resistivity has not been tested using standard laboratory conditions. The aim of this CSM mentor protégé funded project is to implement standard microbiological characterization such as Kirby-Bauer Susceptibility Test to provide qualitative results en par with Minimum Inhibitory Concentration assays to indicate tetracycline sensitivity of the IVYp1 knockout. Assessing these results from Kirby Bauer test, the minimum inhibitory concentrations for the knockout strains will be determined to obtain the minimum amount of antibiotic needed to limit the growth for wild type, IVYp1-, and IVYp2- strains of PA14. Ongoing studies to use complementation of IVYp1 inactivation loop by exogenously added peptide are being used to further indicate the sustainability of IVYp1. IVYp1-/- strain will be examined under the addition of a cyclic peptide consisting of six amino acids followed by MIC assays performed to evaluate if the phenotype TetR is restored through the exogenously added peptide advocating for the necessity and sufficiency of IVYp1 for maintaining the integrity of peptidoglycan cell wall, thus making it a potential drug target.