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Evelyn McChesney & Madeline McCue

Project Summary
Probiotics were engineered to deliver antimicrobial peptides (AMPs) that show promise as an alternative to antibiotics. This work involved designing a digital “blueprint” in SerialCloner for a broad-spectrum Escherichia coli Nissle 1917 AMP delivery system to secrete two AMPs to target enteropathogenic bacterial infections. The AMPs microcin L and enterocin A were chosen because they show antimicrobial activity against gram-negative and gram-positive enteropathogens, respectively. This is the second year of a two-year study.
The first step included designing a plasmid (pMK-P+) with a pMK-RQ-Bb backbone and the strong promoter proTeOn+. The second step was to include secretion machinery that would enable to bacteria to secrete the produced AMP. The microcin L secretion machinery was expensive, so the operon from an available plasmid (pHK22) containing the highly homologous microcin V secretion machinery mutated to halt production of microcin V while maintaining production of the microcin V secretion machinery. The mutated microcin V operon was digitally inserted into pMK-P+ to create the pMK-P+-V plasmid. Finally, gene blocks that encode for production of the AMPs microcin L and enterocin A were designed and digitally inserted into the pMK-P+-V plasmid.
This work is an important step in engineering a broad-spectrum AMP delivery system that has activity against multiple species of enteropathogenic bacteria. Successful synthesis of such a broad-spectrum probiotic would be a tremendous breakthrough in developing an alternative to antibiotics.
 Evelyn McChesney

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   Madeline McCue

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Honors & Awards
Evelyn McChesney (2016) & Madeline McCue (2016) worked in the Department of Chemical Engineering and Materials Science at the University of Minnesota under the guidance of Dr. Yiannis Kaznessis. Maddy and Evelyn won a Second Grand Award at Intel ISEF and presented as national finalists in the Siemens Competition. At the regional science fair they qualified as ISEF alternates, but were named ISEF finalists at the state level. Also at state, Evelyn and Maddy won a First Place Grand Award and the Seagate Rising Star Award for the top high school project. Evelyn will be attending Johns Hopkins University, and Maddy will be attending the University of Southern California.