Applications

Basic Sciences Related

Protein Complementation techniques & Biosensors

Think quick ! That's the message that the EPF Lausanne iGEM team wants to convey. The BioPad project is centered around the use of Protein Complementation Techniques to enable in vivo spatiotemporal analysis of biological signals by bacterial biosensors.

But seriously, what does that mean ? Protein complementation is a technique consisting of the association of reporter protein fragments to components of a same macromolecular complex. Upon reconstitution of the macromolecular structure (active state), the unfolded fused reporter fragments are physically brought together to allow their proper folding. This allows the reconstitution of their chemical properties. The most widely used protein complementation techniques are related to fluorescence (GFP, YFP, RFP), bioluminescence (firefly, renilla luciferases), and cAMP production (Adenylyl cyclase). Most scientists use this technique to validate protein interaction in the context of signal cascades or other pathways.

The EPF Lausanne iGEM team distinguishes itself from this train of thought, as our team implemented a novel split fluorescent reporter to assess the spatiotemporal dynamics of bacterial biosensors. The fluorescent protein used by our iGEM team is the split IFP1.4. The IFP1.4 (engineered Infrared Fluorescent Protein) is a fluorescent protein developed early in 2014 by the Michnick Lab¹. The split IFP1.4 is the first of its kind as it is both fluorescent and reversible (most fluorescent proteins are irreversible). The reversibility is possible as its chromophore - biliverdin - is an organic molecule to which the protein binds. Moreover, the IFP1.4 has advantage of having very low background noise as fluorescence in the far-red spectrum is limited.

The bacterial biosensor allowing the demonstration of our idea was a stress responsive two component regulatory system: the CpxA-R pathway. Our team successfully showed that spatiotemporal dynamics of the biosensor was possible upon fusion of split IFP1.4 fragment to the relay protein of the pathway, CpxR.

Microfluidics as an interface for the in vivo study of Biosensors

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Relationship between genes and their corresponding activating signals

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Applied Sciences Related

Cheap, fast, efficient, and accurate antibiotic screening system

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Antibiotic Complement

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Tumor progression evalutation

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References

1: Michnick, S., Tchekanda, E., & Sivanesan, D. (2014, April 20). An infrared reporter to detect spatiotemporal dynamics of protein-protein interactions. Nature Methods, 6-6.