The project has been divided into several modules - the blue light inducible system, the negative feedback loop, the mutation assay and the lead biosensing and bioremdiating module. Each module is being seperately characterized and then can be later incorporated to gether to form an inducible mutagenesis system.
The results displayed on this page are only from the wet lab experiments. To view the modelling results go here
The overall system has again been divided into two parts. The first part shows light inactivation due to the presence of the fusion protein YF1 and its effector protein FixJ, which in the absence of blue light activates the FixK2 promoter as described in Möglich, Moffat and Ayers[1]. This part is available in the registry as BBa_K3285000.
The second part is a regulatory system with the constitutively active promoter CIlam. The production of RFP is regulated by the cIts2 repressor. The cIts2 repressor is a modified cI repressor which is temperature-sensitive as described here by Jana et al. The cIts2 repressor is present in the repository as BBa_K3285001 and the whole part is present as BBa_K3285002.
The Biosensor Module is mainly involved in quantifying Lead in the system using fluorescence as the readout. In the biosensor, Lead ions in the system prevent a repressor protein (pbrR) from binding to an operator site. GFP is present downstream of the operator site. Therefore we get higher GFP intensities for higher lead concentrations. This part is available in the registry as BBa_K1758333.
