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Team:Bielefeld-Germany/Results/Summary
From 2011.igem.org
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'''S-layer subproject''': 4 different S-layer BioBricks with different lattice structures were created and sent to the partsregistry. The behaviour of these genes when expressed in ''E. coli'' were characterized and purification strategies for the expressed proteins were developed. Two purified fluorescent S-layer fusion proteins from different organisms were immobilized on beads, leading to a highly significant fluorescence enhancement of these beads (p < 10<sup>-14</sup>). | '''S-layer subproject''': 4 different S-layer BioBricks with different lattice structures were created and sent to the partsregistry. The behaviour of these genes when expressed in ''E. coli'' were characterized and purification strategies for the expressed proteins were developed. Two purified fluorescent S-layer fusion proteins from different organisms were immobilized on beads, leading to a highly significant fluorescence enhancement of these beads (p < 10<sup>-14</sup>). | ||
| - | '''NAD<sup>+</sup> detection''': | + | '''NAD<sup>+</sup> detection''': We were able to utilize NAD<sup>+</sup>-dependent DNA ligase from ''E. coli'' (LigA) for a highly sensitive molecular beacon based bioassay detecting NAD<sup>+</sup> in nano molarity scale. The deadenylated form of LigA was able to ligate a split target hybridized to a molecular beacon resulting in an increase of fluorescence intensity which was still measurable in presence of 5 nM NAD<sup>+</sup>. Relating to this, the initial velocity of DNA ligation showed a linear dependence on the employed NAD<sup>+</sup> concentration as long as this remained the limiting factor. |
Revision as of 20:31, 20 September 2011
Bisphenol A subproject: We enabled E. coli to degrade BPA in vivo and improved the specific BPA degradation rate by creating an Fdbisd:CYPbisd fusion protein, changing the cytochrome P450 electron transport system from a putidalike bacterial class I type to a class V type.
S-layer subproject: 4 different S-layer BioBricks with different lattice structures were created and sent to the partsregistry. The behaviour of these genes when expressed in E. coli were characterized and purification strategies for the expressed proteins were developed. Two purified fluorescent S-layer fusion proteins from different organisms were immobilized on beads, leading to a highly significant fluorescence enhancement of these beads (p < 10-14).
NAD+ detection: We were able to utilize NAD+-dependent DNA ligase from E. coli (LigA) for a highly sensitive molecular beacon based bioassay detecting NAD+ in nano molarity scale. The deadenylated form of LigA was able to ligate a split target hybridized to a molecular beacon resulting in an increase of fluorescence intensity which was still measurable in presence of 5 nM NAD+. Relating to this, the initial velocity of DNA ligation showed a linear dependence on the employed NAD+ concentration as long as this remained the limiting factor.
