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E-mail Response:Updating the Progress in the Project
Great to hear from you, but gutted that you haven't got those parts yet. If there's anything we can send you that doesn't involve DNA (e.g. characterisation data) please let us know.
Things are looking up here. We've managed to get powerful blue light from our luxAB-lumP fusion and our red mutants are very very red. We can probably send you pictures soon. We're currently trying to characterise them but we're having issues with our spectrophotometer.
Are Cambridge being helpful? We talked to them in July when they hadn't even started lab work asking if they'd like to collaborate
We're thinking of sticking with the wild type luciferase for green light production just to cut down on time, but if you like we can fuse your YFP with our luxAB and send you the photos and data we get. We'll be crediting you for any parts which you had synthesised and sent to us, including Ccas and YFP, even if you didn't have time to characterise them. I'm sure Chris and the rest of the team wouldn't begrudge doing this for you after all you've done for us. (Having said that, I will have to double check this with Chris before we go ahead, but I'm sure he won't mind.)
We're doing preliminary tests with the red and blue light sensors at the moment, which basically involves leaving one plate in the light and the other in the dark and seeing if there's a colour change. RLS cells should grow blue in the dark and LovTap cells should grow red in the dark. Marta will get back to you about the trp mutants, but we've found it's not that important to have the endogenous trp knocked out. Lausanne didn't when they built the construct, and their's seemed to be fine.
That's all that's going on here really. Lectures, jobs, etc keep stealing our lab time but we're managing. I really hope the parts come through in time for you to work with them before the wiki freeze. Best of luck guys, and don't forget to shout if you need anything.
Maria and the Edinburgh iGEM team.
Updating the Progress in the Project
How are you? How is all going with your project?
We have been very busy and are still waiting for experimental results to test some parts.
We have been focused in firefly and click beetle luciferases because we were unable to ligate LuxAB genes to any plasmid, however we have not detected any bioluminescence yet. Mariana has been in touch with Cambridge team in order to get some advices and also some parts; she is now planning an experimental test using the luminometer.
About the light receptors, the blue promoter ligation with GFP has taken so much time, and Jorge is still verifying if he has obtained it correctly. LovTAP is ready ligated to promoters, but we are still working in the construction of the reporter system (trpL promoter+ GFP/RFP and trpL promoter+ cI repressor +GFP/RFP). So, we don´t know yet if our LovTAP is functional. Have you ever tested it? We are very curious about if it works correctly.
About the trpR mutant strain we would like to know if you have obtained more details about the genotype; in a previous e-mail I mentioned that analyzing the sequence of our mutant, we found a non synonymous mutation (Alanine -> Proline) corresponding to the aminoacid 80.
As maybe you know, we haven’t received your shipment yet (our Customs is awful), so we can´t test other parts for which we need those that you kindly sent us. We really hope that they arrive on time.
Luis has been in touch with your advisor Dr. Chris, he had given us some valuable advices about the measurements in the fluorimeter and also he has helped us with the shipment issues.
Well, we are sorry for the long period without communication and hope to hear news from you.
Good luck during these last weeks before Jamboree.