Butanol biosensors have been developed in previous studies (Dietrich et al., 2013; Yu et al., 2019) but have fallen short in terms of their reproducibility and inducibility. This work aims to characterize existing sensors with a series of butanol inductions and to improve the existing butanol biosensors with a wide variety of synthetic biology tools and techniques. More specifically, this study aims to design and implement synthetic promoters for use in both the biosensors and for general genetic circuit design. Possible promoters were discovered by validating a transcriptomic data meta-analysis of genetic expression levels of Pseudomonas putida under a wide range of stressful conditions with qPCR. One possible promoter was tested in a reporter assay, and another was identified through comparing genetic loci in P. putida to analogous loci in E. coli. In conclusion, degradation of a vital activator protein was observed in the butanol biosensor. The promoter tested with a reporter assay did not perform as expected. This work confirms the need to validate all aspects of a microbial biosensor circuit to ensure its proper function, and to validate meta-analysis results with benchtop experiments to ensure transcriptomic information is transferrable to genetic circuit design.

Creator

• Bruno, Matthew

Contributors

• Vidali, Luis
• Farny, Natalie
• Shell, Scarlet
• Young, Eric

Degree

• MS

Unit

• Biology & Biotechnology

Publisher

• Worcester Polytechnic Institute

Identifier

• etd-63406

Keyword

• Biosensor
• Pseudomonas
• Butanol
• Promoter
• Synthetic Biology
• Gene Expression

Advisor

• Farny, Natalie

Committee

• Young, Eric
• Vidali, Luis
• Shell, Scarlet

Defense date

• 2022-04-12

Year

• 2022

Date created

• 2022-04-25

Resource type

• Thesis

Rights statement

• In Copyright

License

• Creative Commons BY-NC Attribution-NonCommercial 4.0 International