Team:Bielefeld-Germany/Data Page

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[[Image:Bielefeld_2011_S-Layer-Produktion_lysis-purification_v1.jpg|center|700px|thumb|'''Fig. 1: Production of the S-Layer fusion proteins in E. coli.''' To build a cellfree Bisphenol A biosensor the S-Layer fusion proteins have to be extracted from the cells and purified (simplified schema).]]
[[Image:Bielefeld_2011_S-Layer-Produktion_lysis-purification_v1.jpg|center|700px|thumb|'''Fig. 1: Production of the S-Layer fusion proteins in E. coli.''' To build a cellfree Bisphenol A biosensor the S-Layer fusion proteins have to be extracted from the cells and purified (simplified schema).]]
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[[Image:Bielefeld_2011_Bead-Coating_v1.jpg|center|700px|thumb|'''Fig. 2: Coating of the silica beads with two different S-Layer fusion proteins.''']]
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[[Image:Bielefeld_2011_Bead-Coating_v1.jpg|center|700px|thumb|'''Fig. 2: Coating of the silica beads with the S-Layer fusion proteins.''' Every silica bead gets covered by a geometric S-Layer film consisting of an alternating structure of the two essential S-Layer fusion proteins.]]
[[Image:IGEM_Bielefeld_Project.jpg|center|700px|thumb|'''Fig. 3: Visualization of our cellfree Bisphenol A biosensor 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<sup>+</sup> dependent ligase (LigA) ligates the two oligos so that the hairpin structure opens up and the fluorophore is able to emit light.]]
[[Image:IGEM_Bielefeld_Project.jpg|center|700px|thumb|'''Fig. 3: Visualization of our cellfree Bisphenol A biosensor 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<sup>+</sup> dependent ligase (LigA) ligates the two oligos so that the hairpin structure opens up and the fluorophore is able to emit light.]]

Revision as of 17:55, 17 September 2011

Contents

How our System works

Fig. 1: Production of the S-Layer fusion proteins in E. coli. To build a cellfree Bisphenol A biosensor the S-Layer fusion proteins have to be extracted from the cells and purified (simplified schema).
Fig. 2: Coating of the silica beads with the S-Layer fusion proteins. Every silica bead gets covered by a geometric S-Layer film consisting of an alternating structure of the two essential S-Layer fusion proteins.
Fig. 3: Visualization of our cellfree Bisphenol A biosensor 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. [http://partsregistry.org/Part:BBa_K525305 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. [http://partsregistry.org/Part:BBa_K525517 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. [http://partsregistry.org/Part:BBa_K525710 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. [http://partsregistry.org/Part:BBa_K123000:Experience 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. [http://partsregistry.org/Part:BBa_K123001:Experience 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. [http://partsregistry.org/Part:BBa_K525405 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. [http://partsregistry.org/Part:BBa_K525512 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>.
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