Team:MIT/Projects

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(Project 2: Rapid & Reversible Protein Localization using PhyB-PIF3 System)
(Project 2: Rapid & Reversible Protein Localization using PhyB-PIF3 System)
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Our goal is to engineer a system that adopts the PhyB-PIF3 switch to control protein localization within the cell. We plan to have either PhyB or PIF3 constitutively anchored to the target desired (e.g. mitochondrial membrane, nuclear membrane, vacuole etc). The other will then be bound to our protein of interest(fluorescent protein) and will diffuse within the cell. When pulsed with red light, the traveler protein will localize to the anchored protein.<br>
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Our goal is to engineer a system that adopts the PhyB-PIF3 switch to
 +
control protein localization within the cell. In our design, either
 +
PhyB or PIF3 is constitutively anchored to one of the four target
 +
locations. The other will then be bound to our protein of interest and
 +
diffuse within the cell. Here, we use CFP and YFP to track the
 +
location of PhyB and PIF3, respectively. Reversable, light-dependent
 +
association of PhyB and PIF3 can then be monitored by fluorescence
 +
microscopy.<br>
[[Team:MIT/Projects/Project2|Link to more details about Project 2]]
[[Team:MIT/Projects/Project2|Link to more details about Project 2]]
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Revision as of 00:45, 22 October 2009


Home|Projects|Protocols|[http://partsregistry.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2009&group=MIT Parts for Registry]|References

Project 1: Metabolic Engineering of PCB Synthesis in Yeast


PCB factory.jpg

Brief Description


Our project uses the PhyB-PIF3 system to create this fast-reversible switch. Phycocyanobilin (PCB) is a chromophore necessary for this switch to work. PCB is the protein that senses light and changes its conformation to allow PhyB to bind to PIF3, as well as unbind. In its inactive conformation, noted as the Pr conformation, can absorb red light and change its conformation to the active conformation, noted as the Pfr conformation. In the active conformation, PCB can absorb far-red light to revert back to the inactive conformation. We wanted to make this process self-sufficient. Therefore if the yeast was able to synthesize the PCB itself, it would not have to always be supplemented.
Link to more details about Project 1

Project 2: Rapid & Reversible Protein Localization using PhyB-PIF3 System


Localization system.jpg

Brief Description


Our goal is to engineer a system that adopts the PhyB-PIF3 switch to control protein localization within the cell. In our design, either PhyB or PIF3 is constitutively anchored to one of the four target locations. The other will then be bound to our protein of interest and diffuse within the cell. Here, we use CFP and YFP to track the location of PhyB and PIF3, respectively. Reversable, light-dependent association of PhyB and PIF3 can then be monitored by fluorescence microscopy.
Link to more details about Project 2