Drake Heinz

Date of Award

Spring 5-2020

Document Type


Degree Name

Master of Science (MS)



First Advisor

Patrick Hindmarsh


In today’s world, there is an urgent need to discover novel antibiotics due to the rise of antibiotic resistance and the lack of new ones being approved. Building off of Hanson et al. and their reduction-oxidation sensitive green fluorescent protein (roGFP)(1), that we yeast codon optimized (2); I wanted to determine if I could create a real-time biosensor that has the ability to respond to ROS produced in the stress response triggered by antibiotics as a platform to identify novel antibiotics. To achieve this, I need to know if it will oligomerize in bacterial cytoplasm due to its two cysteines at 48 and 70 and what the optimal conditions are for the biosensor. From western blot results, royGFP does not oligomerize in the cells showing that our biosensor should not lose fluorescence. royGFP had a strong sensitivity to hydrogen peroxide and ROS triggered from antibiotics. The results showed that royGFP can be used as an ROS biosensor tool to help discover novel antibiotics.