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Design and Partial Assembly of a Promoter Characterization Circuit for Mycobacteria
Nafziger, Elias H.
Nafziger, Elias H.
Abstract
Mycobacterium is a diverse genus of gram-positive, aerobic bacteria known for their waxy cell membrane, resilient biofilms, and harmful pathogens. Bioengineering has enabled their use in a variety of fields. Mycobacteria have been engineered as vaccines and to gain insight into their pathogenesis. Species like Mycobacterium neoaurum are currently the only bacteria used at the industrial level to synthesize sterols for various therapeutic uses. Finally, mycobacteria in soil have been shown to break down highly stable pollutants, while their aquatic counterparts have been suggested as “probiotic” coatings in plumbing. All of these applications of mycobacteria could be expanded beyond their current state with synthetic biology by promoting rational design, system modeling, and standard genetic part characterization. However, mycobacteria lack the fundamental underpinnings of standard genetic parts to model and assemble genetic circuits with. We have designed and partially assembled a promoter characterization method to characterize promoters in mycobacteria in a standard, highly qualitative manner. The genetic circuit pSUM7 constitutively expresses mCherry while simultaneously expressing sfGFP, which is regulated by a test promoter. Ultimately, we plan to generate a comprehensive promoter library from which synthetic biologists can select promoters a la carte that best fit their modeling parameters and design goals. This will help lay the foundation for synthetic biology in mycobacteria and rational circuit assembly.
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2022-05-01
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5/10/2032
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Biology