Kristian M. Müller, Katja M.Arndt
Andreas Hiltbrunner, Michael Reth, Bodo Rak
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-Calmodulin-DHFR construct.
final iGEM report: Ca2+ sensor
final iGEM report: light sensor
Biobrick compatible strategy for fusion proteins
The present BioBrick prefix and suffix rules are not compatible with modular protein design. Thus, we propose an extension of the present standard for fusion proteins in which two restriction sites are added in frame adjacent to the coding sequence.
final iGEM report: Fusion parts
Mediums and Plates
Plasmid spin column prep
General Gene-Protein Information
Polyacrylamide gel electrophoresis
In vivo test I
In vivo test II