Team:LMU-Munich/Data/MazF
The LMU-Munich team is exuberantly happy about the great success at the World Championship Jamboree in Boston. Our project Beadzillus finished 4th and won the prize for the "Best Wiki" (with Slovenia) and "Best New Application Project".
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Cloning of the toxin MazF in E. coli
The cloning of the toxin MazF in E. coli itself was tricky. Even though the MazF itself worked, it didn't grow well, and after retransformation we got only six colonies to pick (right picture above). This shows that this Biobrick is probably functional and is a problem for E. coli to handle even without promoter.
The 3A assembly with the promoter did not work at all (left picture above). We yielded mostly just red colonies and the few that were white were wrong, never having the promoter with MazF. After many attempts with either the pSB1C3 or our own Bacillus vectors, we chose to directly transform the 3A assembly into Bacillus subtilis in hopes that it would cope better with the toxin and/or the promoter would be less leaky. This was too late to finish before Amsterdam unfortunately. But maybe the story will be told to an end at Boston.