| Types | DnaRegion
|
| Roles | engineered_region
Reporter
|
| Sequences | BBa_K621001_sequence (Version 1)
|
Description
Our part consists of an OMP-R binding region that regulates our reporter gene, eYFP. The OMP-R binding region is an omp-c operator, which has three OMP-R binding sites. Our part is regulated by the ENVZ-OMPR two-component regulatory system. ENVZ is a osmolarity sensor on the inner membrane and phosphorylates OMP-R in the presence of high osmolarity. When phosphorylated, OMP-R binds to the omp-c operator and recruits RNAP to transcribe the downstream reporter gene. Fluorescence intensity due to the production of eYFP is a measure of the system's activity. When all three binding sites are occupied by phosphorylated OMP-R proteins, RNAP cannot bind to the DNA and transcribe downstream. This part can be used as an osmolarity indicator. Our team used this part as a glucose sensor because the presence of glucose changes osmolarity. Different concentrations of glucose produce different, quantifiable fluorescence intensities.
Notes
Our team had to combine the genes in a specific order. First our team cut out and purified the eYPF gene. We added this gene to a plasmid containing RBS. Next we cut out the RBS+eYFP intermediate and ligated them into the plasmid containing OMP-R. The RBS and OMP-R parts were too small to run out and extract on a gel because they would run off of the end of the gel. Therefore, we had to start the process with eYPF.
Source
This BioBrick was created from parts found in the Registry. These parts were BBa_R0082, BBa_B0032, and BBa_E0030.
