Inducibility of Synthetic Genetic Circuits in Pseudomonas putida under Simulated Soil Conditions

Abraham, Lauren

Student Work

Inducibility of Synthetic Genetic Circuits in Pseudomonas putida under Simulated Soil Conditions

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This research tests the efficacy of an inducible system present in genetically engineered bacteria Pseudomonas putida (P. putida), a widespread soil bacterium. Inducible systems in bacteria are used for a variety of functions in biotechnology, including bioremediation and working as cell factories to produce natural products and small molecules. The inducible system used in this research responds to the addition of an anhydrotetracycline (aTc) with the synthesis of the far-red fluorescent protein mCardinal to report the presence of aTc. To further research and address how the biosensor would respond under real-world environmental conditions, this Major Qualifying Project expanded upon biosensor testing under simulated soil conditions using a liquid soil extract in the lab for testing. The goal of this research project was to observe the effects, if any, of addition of an aTc inducer to the inducible system in liquid soil extract. While we were able to confirm inducibility in laboratory growth conditions, we were unable to induce the circuit in liquid soil extract conditions. We believe that the promoters that drive the system may not be functional under soil-like conditions and will need to be further optimized in order to apply inducible genetic circuits within real-world environmental conditions. These discoveries motivate further research into the potential use of P. putida as a biosensor for antibiotics and other environmental contaminants in soil environments.

This report represents the work of one or more WPI undergraduate students submitted to the faculty as evidence of completion of a degree requirement. WPI routinely publishes these reports on its website without editorial or peer review.

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