Part:BBa_M50026
Gold Detection Device with Promoter RBS and GolS
In this paper, we designed a sensor/reporter device, pGold. These parts include a promoter that binds a GolS inducer, a ribosome binding site (RBS), and a gene coding for GolS, a protein originating from Salmonella typhimurium that forms a protein-metal complex with gold (Au). GolS, upon binding Au, dimerizes and then binds a recognition site on the promoter, forming the basis for our sensor device.
The pervasiveness of bacteria throughout the body presents an opportunity to engineer our microbiome to express novel therapeutic biologics. In our study, we use Au3+ gold salt as an analogue for gold nanoparticles (or other metals usable in metal-inducible expression cassettes) to bind to GolS and form a protein-metal complex which serves as an activator for the promoter region which will increase expression of GFP downstream. To validate our design, we run a series of fluorometric tests to quantify GFP output and cell growth/density over a 16 hour time period. Our findings serve as a proof-of-concept, demonstrating that a gold-inducible expression plasmid can be synthesized and transformed into bacterial cells to upregulate expression of genes of interest under certain Au3+ concentrations. However, since Au3+ is a charged molecule and cannot be safely administered into the body, our future works centers around designing a transcriptional activator that can detect uncharged biocompatible gold-nanoparticles. The GolS protein must be modified to selectively bind to uncharged-gold nanoparticles.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 485
- 1000COMPATIBLE WITH RFC[1000]
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