From 2011.igem.org

Revision as of 17:35, 17 September 2011

How our System works

Fig. 1: Schematic drawing of the S-Layer fusion protein production in E. coli with cell lysis and purification.

Fig. 2: Coating of the silica beads with two different S-Layer fusion proteins.

Fig. 3: Visualization of our cellfree Bisphenol A sensor system with all essential components. Bisphenol A (BPA) is reduced by the electrons from NADH transfered by reductase (Red), ferredoxin (Fd) and cytochrome P450. The molecular beacon (hairpin structure) binds two short DNA-oligos. The NAD⁺ dependent ligase (LigA) ligates the two oligos so that the hairpin structure opens up and the fluorophore is able to emit light.

Data For Our Favorite New Parts

1. Main Page - Fusion Protein of S-Layer SgsE and mCitrine: This fluorescent S-layer fusion protein is used to characterize purification methods and to demonstrate the S-layer's ability to self-assemble on surfaces.
2. Main Page - Fusion Protein of BisdA and BisdB (expressed): This fusion protein improves the bisphenol A degradation in E. coli compared to the so far in the partsregistry existing BPA degrading BioBricks.
3. Main Page - NAD⁺-dependent DNA ligase from E. coli : This enzyme enables determination of NAD⁺ even in very low concentrations by coupling it with a molecular beacon based assay.

Data For Pre-existing Parts

1. Experience - BisdA degrades Bisphenol A when used with BisdB, BBa_K123000 (University of Alberta, iGEM 2008): Complete degradation of 120 mg L^-1 Bisphenol A with polycistronic bisdAB gene in 30-33 h. Even faster (21-24 h) when using a fusion protein of BisdA and BisdB.
2. Experience - BisdB degrades Bisphenol A when used with BisdA, BBa_K123001 (University of Alberta, iGEM 2008): Complete degradation of 120 mg L^-1 Bisphenol A with polycistronic bisdAB gene in 30-33 h. Even faster (21-24 h) when using a fusion protein of BisdA and BisdB.

We've Also Characterized the Following Parts

1. Main Page - Fusion Protein of S-Layer SbpA and mCitrine: This fluorescent S-layer fusion protein is used to characterize purification methods and to demonstrate the S-layer's ability to self-assemble on surfaces.
2. Main Page - Polycistronic expression of BisdA and BisdB: This is the version of BPA degrading BioBricks found in the partsregistry - comparison to our fusion protein <partinfo>K525515</partinfo>.