Notebook:Federico Castro M/Projects/Repressilator
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Revision as of 15:51, 19 January 2008
Last updated January/17/2008
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Project status: Active
In order to improve lab skills and to see with my own eyes the repressilator I intended to assemble it with the biobricks available at 2007 part kits.
At first I tough I could build the repressilator with 3 inverters, one protein generator and one single part... then I realized that almost all the devise was already assembled into one huge biobrick DUH! While searching within the registry I found that only EYFP had the correct degradation tag; other fluorescent proteins had a tag that was to strong for this devise or had no tag at all.
It seems that I will have to use EYFP (I kind of dislike EYFP) so the final construction will be exactly like <bbpart>BBa_I5611</bbpart> O_o Even when the final construction is already available the biobricks obtained and the practice should be worth the time and hard work. Besides, the whole process should be very simple and It shouldn't take more than a week of work.
Now I only have to transform <bbpart>BBa_S03151</bbpart> and <bbpart>BBa_E0430</bbpart> and assemble them.
Alternatively I could use the parts <bbpart>BBa_I5612</bbpart> and <bbpart>BBa_I13602</bbpart>.
I don't know why there are so few GFPs with LVA tags at the registry. Elowitz used a GFP instead of a EYFP. I'm not such a purist so I will make my own devise devise with EYFP in the plasmid pSB1A2 and I wont use IPTG (anyway Elowitz was unable to synchronize the cells with IPTG).
Have you seen the zize of I5612? It's 3337 bp and thats a lot for a plasmid and yet I want to add some 886 bp... I wonder if that would burden the plasmid so much that it would be unable to replicate along with the bacteria yet the whole construction with 4223bp (<bbpart>BBa_I5611</bbpart>) is available. There is only one way to know if a plasmid can hold 4000+ bp hahahaha.
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