Team:MIT/Projects/Project1
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<center><math>C(mM)=\frac{A_{680}}{37.9}*\begin{cases} 100 (for 1:100) \\ 1000 (for 1:1000) \end{cases}</math></center> | <center><math>C(mM)=\frac{A_{680}}{37.9}*\begin{cases} 100 (for 1:100) \\ 1000 (for 1:1000) \end{cases}</math></center> | ||
- | ==Engineering the PCB Synthesis Pathway into Yeast== | + | ===Engineering the PCB Synthesis Pathway into Yeast=== |
[[Image:PCB_Biosynthesis_Pathway.png|center|500px]] | [[Image:PCB_Biosynthesis_Pathway.png|center|500px]] | ||
<center>''Adapted from .....''</center> | <center>''Adapted from .....''</center> |
Revision as of 01:19, 20 October 2009
Metabolic Engineering of PCB Synthesis in Yeast
As shown in the image above, phycocyanobilin (PCB) plays a crucial part in the PhyB-PIF3 system. Once PCB is in the active conformation it allows the phytochrome (PhyB) to bind to the integrating factor (PIF3).
Developing the Standard: PCB from Spirulina
We decided to use a standard used in many other experiments involving phytochromes. Phycocyanobilin (PCB) extracted from Spirulina is a commonly used standard, as Spirulina produces a large amount of chromophores. We used Spirulina which was bought at Vitamin World as it is commonly used as a dietary supplement.
Chromophores have a very high absorbance around 680nm. Here is an example of an absorbance spectrum:
We followed the protocol that the Quail Lab used to extract the PCB from Spirulina, and were able to produce the following spectrum:
The concentration is found by this formula: