Backbone

To get our developed biobricks into b. subtilis, a backbone was developed. The biobrick compatible backbone developed by the iGEM team of Munich 2012 (Insert exact name) was used as an template but is improved (insert link to part registry) . A hyspank promoter was added to the backbone. The multiple cloning site is used to enter our biobricks into the system. For the use of b. subtilis. the amyE sites are used to cut it

We used the empty backbone for integration into Bacillus subtilis amyE locus from Munich 2012 (BBa_K823023) that had no promoter. We've added the Hy_Spank IPTG inducible promoter from Rudner's Lab 2004. The Hy_Spank promoter is right in front of the prefix so any biobrick can be inserted in this backbone.
We used the empty backbone for integration into Bacillus subtilis amyE locus from Munich 2012 (BBa_K823023) that had no promoter. We've added the Hy_Spank IPTG inducible promoter from Rudner's Lab 2004. The Hy_Spank promoter is right in front of the prefix so any biobrick can be inserted in this backbone.

Promoter activity

The intensity in of ~40 cells in the GFP-channel, were analyzed from two pictures.
The average intensity (AU) from the cells are plotted above with the standard deviation.

GFPmg and GFP (BBa_E0840)