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Team:METU Turkey/characterization.html
From 2013.igem.org
We have done the characterization of three promoters as models. RFP genes were transcribed using these promoters and the amount of RFP molecules were modeled by considering their brightness. The results are as follows:
1- K606040: Promoter Hyperspank B. subtilis & E. coli (iGEM11_Paris_Bettencourt)
We simulated the production of RFP under two different IPTG concentrations to observe its basal and maximum transcription rates.
As seen in the graph, when there is no IPTG with the increasing LacI expression, the RFP amount becomes constant and this means there is no more production of RFP.
As seen in this graph, with 5000 IPTG molecules, LacIs form complexes with IPTG. Therefore there is decreased inhibition on K606040 and RFP production continues and the amount keeps getting higher.
2- K823003 Pveg (iGEM12_LMU-Munich)
We did alignment by using CLUSTAL W between two Pveg promoters; K143012 and K823003. The result is as follows:
K823003 sequence has 237 bp and K143012 sequence has 97 bp as seen in the figure. We can conclude that K823003 includes K143012. After BLASTing the remaining part of the K823003 which is not same as K143012 there were no results for specific sequences for polimerase binding. This shows K823003 has alike promoter strength as K143012. Only the primers were desinged further from the main promoter region. So for this promoter we can use the promoter strength of the K143012.
3- K143012 Promoter veg: Constitutive Promoter for B. subtilis (iGEM08_Imperial_College)
We used known parameters (parameter strength=0.79 rpu) of K143012 to simulate RFP production by using this promoter.
This graph shows constitutive production of RFP by using K143012.
