Team:INSA-Lyon/Biology

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Curly'on - IGEM 2014 INSA-LYON

IGEM

WETLAB SUMMARY

Our wetlab work focuses on designing a bacterial strain able to chelate as much Nickel as possible but also to adhere to a synthetic matrix for future filter design applications. To do so, we engineered a hair-shaped protein polymer located at the bacterial surface, called curli (SOURCE). CsgA, which is the monomer of the Curli structure, can be engineered by our will. This property constitutes the basis of our work because we engineered the CsgA by adding one or more His-Tag motifs, famously known to be able to chelate Nickel (SOURCE). Our project aims at modifying an Escherichia coli strain that naturally produces abundant biofilm and at the same time produces the engineered curli proteins to helate Nickel. This way, the bacteria have the sufficient adhesion ability to stick to the filter matrix and be exposed to polluted water and chelate the environmental nickel.

  • As a first step, we constructed and cloned a modified CsgA that has either one or two His-Tag motifs. This way, we will be able to investigate if a repeated His-Tag motif is able to chelate more Nickel or not.
  • As a follow-up, we designed and optimized two protocols for nickel chelation investigation: one based on post-centrifugation supernatent exposure to nickel and a second one based on curli purification only. Both technics use dimethylglyoxime (DMG) that changes color from transparent to bright red in the presence of Nickel.
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