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When we discovered that we did not have the desired Biobricks in the registry, we decided to design and characterize our own Biobricks. We wanted to characterize the superfolded GFP which was optimized for Lactococcus lactis and the CP promoter collection. We made the constructs containing the CP promoter collection with the super folded GFP. Although the superfolded GFP was optimized for L. lactis, it works in E.coli as well. The promoter collection was studied in E.coli by measuring the fluorescence in FACS. Based on the FACS data we analyzed the promoter activity of the CP promoter collection and proved superfolded GFP works well in E.coli too. One of our new Biobricks, NisA, was characterized by putting it before a CP44 promoter. NisA was then transformed to Lactococcus lactis and a nisin activity test was done. But unfortunately we could not see any zone of inhibition.
Team:Groningen/Template/MODULE/project/characterisation
From 2014.igem.org
Project
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Characterization
Data
Figure 1:
The graph shown above helps us to know distribution of the fluorescence in the cells of E.coli with different CP promotor fused to superfoldedGFP-terminator-RBS collection (CP1, CP11, CP29, CP30, CP41, CP44) and J23101 as reference
Collaboration
Figure 2:
LMU Munich https://2014.igem.org/Team:LMU-Munich helped us to characterize our CP promotor collection in Bacillus subtilis. Unfortunately this yielded no results
