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Team:CSU Fort Collins/Desalination
From 2013.igem.org
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Desalination
The Concept
Near the ocean but you don't have any water to drink? Have no fear, desalination is here! The CSU iGEM team has a great concept for you to use that just might come to a store near you! Simply insert our tea bag, that contains our special yeast, into a glass of sea water and within minutes you'll have salt free water! That's not all though, take your tea bag and use it to power a light bulb or something like that...
The Theoretical Mechanism
This was the ambitious project. This project involved attempting to create a yeast that would sequester sodium ions into vacuoles, removing them from the surrounding environment.
A) one part was to modify an endogenous high capacity sodium transporter (NHA1) so that it would be redirected from the plasma membrane to the yeast vacuole
B) a second part involved the modification of an endogenous Na transporter (NHX1) present on the vacuole so that it would be over expressed, enhancing the rate of sodium movement
C) a third part was creating a yeast strain with all plasma membrane Na- ATPases (ENA1, 2 and 5)knocked out, preventing the yeast from expelling the yeast back into the environment, a normal wildtype behavior
D) and the forth part was to design a channel rhodopsin that would allow the system to be light activated, only allowing Na to enter the cell when exposed to a specific wave length of light.
The design of the first component (part A) of this project involved identifying both plasma membrane and vacuole targeting sequences, and a functional synthetic membrane protein using these sequences had never been created before (to the best of our knowledge).
Results
Our plan was to design our bio-bricks using Gibson Assembly and gBlocks. However we were unable to get the Gibson Assemblies to work, and we were unable to produce our parts. One big hurdle in designing our parts were that some of the sequences for the proteins we wanted to use had many restriction enzyme sites that were not compatible with the bio-brick standards. This is why we chose Gibson Assembly of gBlocks over PCRing our sequences out of the yeast genome.
