Freiburg

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Revision as of 15:44, 20 October 2007

Team Members:

Picture: The University of Freiburg iGEM Team in June 2007 (left to right: Natalia, Dinah, Corinna, Philipp, Katja, Maximilian, Moritz, Dario, Kristian)

Instructors:

Kristian M. Müller, Katja M.Arndt

Students:

Maximilian Mauler

Moritz Busacker

Corinna Gruber

Natalia Maier

Dario Hermida Aponte

Philipp Mappes

Graduate Student:

Dinah Mattay

Advisors:

Andreas Hiltbrunner, Michael Reth, Bodo Rak

Project Summary

Integrated Sensor-Executor Proteins and Molecular Switches
Our goal is to design integrated molecular sensing and executing devices based on modular protein engineering. These integrated devices can then easily be used for the construction of complex systems. We fuse sensing proteins, which provide nano-mechanical movements or dimerization upon an external signal, to executing proteins, which depend in their activity on the nano-mechanical change in the sensing part.
To elucidate the possibilities of such a system we used the calcium-ion sensor Calmodulin and the light sensor system PhyA-Fhy1. To test execution we used the split enzymes DHFR or beta-lactamase or the fluorescent proteins CFP and YFP, which can form a FRET pair. Sensors and executors were geneticlly fused and tesetd in E. coli for activity. So far we could demonstrate Ca2+ dependent growth of E. coli with the DHFR[1]-Calmodulin-DHFR[2] construct.

Biobrick compatible strategy for fusion proteins

Support:

[http://www.syntheticbiology.ethz.ch/synbiocomm/index SYNBIOCOMM]

[http://www.uni-freiburg.de/ Universität Freiburg], [http://www.uni-freiburg.de/wiss-ges/ Wissenschaftliche Geselschaft in Freiburg im Breisgau]

[http://www.geneart.com GeneArt]

Lab-Work:

lab notes
plasmids
PCR primers
Lab Boxes

Protocols:

DNA sequencing
Plasmid spin column prep
Glycerol stocks
General Gene-Protein Information

@@ Line 31: / Line 31: @@
 Andreas Hiltbrunner, Michael Reth, Bodo Rak
-== Our Project: ==
+== Project Summary ==
-Our goal is to design integrated molecular sensing and executing devices based on modular protein engineering. These integrated devices can then easily be used for the construction of complex systems. We plan to fuse sensing proteins, which provide nano-mechanical movements upon an external signal, to executing proteins, which depend in their activity on the nano-mechanical movement of the sensing protein. To elucidate the possibilities of such a system we plan to fuse each of the termini of Calmodulin, which senses Ca2+, or the Maltodextrin Binding Protein, which senses maltodextrin, to one half of a split enzyme such as β-lactamase. We expect that we can design the separation of the two enzyme fragments by one conformation of the sensing protein such that there is no activity and that a conformational change in the sensor will bring the enzyme fragments together resulting in activity of the execution part. Once the basis of the interconnection of sensor and executor has been established, we plan to extend the work in a modular fashion to further proteins in order to generate a readily available and easy to tailor toolbox. We see application of such tools as varied as bacterial growth regulation and tumor targeting.
+<b>Integrated Sensor-Executor Proteins and Molecular Switches</b><br>
+Our goal is to design integrated molecular sensing and executing devices based on modular protein engineering. These integrated devices can then easily be used for the construction of complex systems. We fuse sensing proteins, which provide nano-mechanical movements or dimerization upon an external signal, to executing proteins, which depend in their activity on the nano-mechanical change in the sensing part.<br>
+To elucidate the possibilities of such a system we used the calcium-ion sensor Calmodulin and the light sensor system PhyA-Fhy1. To test execution we used the split enzymes DHFR or beta-lactamase or the fluorescent proteins CFP and YFP, which can form a FRET pair. Sensors and executors were geneticlly fused and tesetd in E. coli for activity. So far we could demonstrate Ca2+ dependent growth of E. coli with the DHFR[1]-Calmodulin-DHFR[2] construct.
+<br>
+<b>Biobrick compatible strategy for fusion proteins</b><br>
 == Support: ==

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