Team:MIT/Projects

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Project 1: Metabolic Engineering of PCB Synthesis in Yeast

Brief Description

Phycocyanobilin (PCB) is a chromophore necessary for PhyB-PIF3 based synthetic devices. Our goal is to engineer a yeast strain capable of synthesizing PCB, so that no exogenous PCB would be needed for PhyB-PIF3 to function in yeast. We thus cloned the genes encoding enzymes in the PCB biosynthetic pathway for expression in yeast
Link to more details about Project 1

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

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

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-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.
+Phycocyanobilin (PCB) is a chromophore necessary for PhyB-PIF3 based
+synthetic devices. Our goal is to engineer a yeast strain capable of
+synthesizing PCB, so that no exogenous PCB would be needed for
+PhyB-PIF3 to function in yeast. We thus cloned the genes encoding
+enzymes in the PCB biosynthetic pathway for expression in yeast
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 [[Team:MIT/Projects/Project1|Link to more details about Project 1]]