Team:Freiburg/4 April
From 2011.igem.org
Attending: Manuel, Sophie, Jabok, Julia, Theo, Rüdiger, Johannes
Tasks
scheme: task(responsible persons)
- Logo-Design, T-shirt, advertising(Sophie?)
- Wesbite/Wiki
- Sponsoring (Julia, Theo, jabob)
- Press releases(Josef, Rüdiger)
- Ethical considerations + Saftey&Security (Josef, Theo)
- Lab and meeting protokols (Julia)
- Prepare Lab methods (Jakob, Sophie)
- purchase(Theo)
- Cloning strategy(Theo)
- Modelling -graphical/mathematical..
- Theoretical planing (Rüdiger: TLR4 Receptor, ...)
- Team leader(Rüdiger)
- Labwork(all except Josef)
- Graphics/Poster
Contact persons:
- Ethics: Boldt Ethikseminar Freiburg
- Logodesign: Lora Braun (toolbox)
- Test licence for lab protocol: Tomasz Krzyzowski BIOSS IT Abteilung.
- Press: BIOSS Christiane Gieseking Anz
- Chemical purchase Andrea Weber BIOSS
Project idea: Cellular Explorer
a system of several light inducible promotors that control genes a molecular biologist needs. The light promotors are saved in the registry: blue, green, red. Tokyo iGEM team 2009(?) developed a counter for light pulses. With this extension, several genes could be controlled by one light source. Genes stored in the "Explorer": Taq-Plymerase, Ligase, Exonuclease (for Gibson Cloning, see Cambridge Team 2010), cell lysis device, cell aggregation device, protein His tag purification device(this is what we will develop)
Protein purification device:
cells express a soluble protein which is able to specifically bind a plastik/paper surface it also includes a LRR or LRRNT motif (http://smart.embl-heidelberg.de/smart/do_annotation.pl?ACC=SM00013, also compare TLR4 studies), maybe combined with a Histidine rich repeat motif. This structure will bind several Nickel ions in close proximity. These Ni ions have free coordination points with which they can bind His tagged proteins. The application could be:
- induce protein purification device by light pulse.
- lyse cells by light pulse.
- centrifuge.
- pour supernatant over paper/plastic surface
- wash off surface - only specific proteins will stay
- induce his tagged protein of interest expression by light pulse
- induce lysis by light pulse
- centrifuge
- pour supernatant over prepared surface - his tagged proteins will bind
- wash of rest
- eluate his tagged proteins with imidazole