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Team:Technion/23_April_2012
Ilya
During the past few days we were learning cloning protocols with Sharon. It is nice to work with the rest of the gang in the lab. I keep saying that no matter what, we learn from mistakes. I will get the cloning right eventually (third time is a charm).
We finally divided into the subgroups that will work on the subprojects. I am in group 1 with Inbal and Asaf. We are goind to construct a new buffer gate that will produce T3 RNA polymerase in response to our chosen input. At first, Roee suggested we use a pLux/Ara promoter, but since group 3 uses a pLac/Ara promoter we need a different solution.
The idea we came up with is to use a pLux promoter along with a theophyllin riboswitch. This way, the riboswitch will lower the basal level of expression. Once we add AHL and theophyllin as our chosen input we will get full induction.
To achieve this, we need to find a strain of E. coli that expresses LuxR or a similar transcription factor in high levels from its chromosome. Then we can construct our buffer gate using a pLux promoter, the theophyllin riboswitch, the T3 RNA polymerase and the fluorescent reporter protein. This will complete our plasmid for stage 1.
Let the search begin!
Inbal
Asaf
Hila
Lior
Noa
Evgeni
Shahar
Rachel
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