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Team:MIT/Projects
From 2009.igem.org
Project 1: Metabolic Engineering of PCB Synthesis in Yeast
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Brief DescriptionOur 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.
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Project 2: Rapid & Reversible Protein Localization using PhyB-PIF3 System
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Brief DescriptionOur 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. |
