Teaching low level programming in the C and ARM assembly languages.

IT work managing Windows and Mac OS based devices using InTune and Mosyle MDMs. Other duties include network management and device installations.

We characterized a cytotoxin that is excreted via a type III secretion system (T3SS) in Pseudomonas aeruginosa, in the lab of Dr. Donald Rowen. The cytotoxin is localized to the membrane bound T3SS via a critical chaperone. The manner in which the chaperone binds the toxin's chaperone-binding-domain is novel, compared to other T3SS effectors which also bind to chaperones. We sought to elucidate details of these interactions. A few of our methods included:1) Creation of mutants which abrogate interactions between critical residues of the chaperone and toxin. 2) Visualization of mutant structure via protein crystallization and subsequent x-ray crystallography.3) Creation of GFP fusion constructs to quantify interaction levels via fluorescence spectroscopy. We will also visualize cellular localization of the mutants and wild type, using fluorescence microscopy.

I studied the genetic basic of bacterial responses to antimicrobial peptides (AMPs). I sought to discover the molecular basis of bacterial resistance mechanisms of two ESKAPE pathogens, by identifying responding genes from transposon libraries. Downstream work involves synthetic construction of AMPs, which specifically target pathways that are exclusive to human pathogens. We also characterized a system involved in mediating resistance, with homologues in S. aureus, E.coli and P. aeruginosa. This work involves the traditional overexpression and knockout experiments, along with mutating key residues. I am concurrently identifying optimal protein purification conditions and doing in silico work on identifying potential inhibitors. Should potential inhibitors be verified (using techniques such as SPR and HPLC), they will be incubated with protein crystals, for use in structure determination via x-ray crystallography.Other duties included supervising graduate students and guiding research technicians.

I was primarily involved with performing quality control of OTC products via HPLC analysis, along with conducting validation study experiments, and editing validation reports.

I worked on many projects during my PhD career. Just a few of the highlights include the following:1) Characterization of Cid/Lrg proteins from Staphylococcus aureus- The membrane-bound Cid/Lrg proteins are involved in biofilm formation, and are controlled by a transcriptional regulator, CidR. I helped express and purify these proteins, as well as perform in silico analysis to aid in crystallization.2) Made fusion constructs of proteins with either N-terminal or C-terminal domains of GFP, to visualize cellular localization of Cid-Lrg dimers, using fluorescence microscopy. Quantitation was also performed using fluorescence spectroscopy.3) Successful crystallization and structural determination of the type II Dehydroquinase from Pseudomonas aeruginosa. Enzyme kinetics assays were performed on it, and a homologue from Francisella tularensis. Potential inhibitors were chosen based on structural features and in silico predictive algorithms, and subsequently tested in inhibition assays. 4) Creation of constructs with a membrane protein of interest fused to an antiviral lectin, to improve solubility of such proteins, which normally have poor yields using traditional membrane protein purification techniques.