1. Constructed BioBricks
Our project required the construction of fusion proteins that are displayed on the surface of the spores of B. subtilis. Anchoring an epitope-specific nanobody and glutathione S-transferase to coat proteins enables the spores to attach to surfaces and activate prodrugs on site. In addition to targeted drug delivery, site specific enzymatic activities play a crucial role in other scientific field and many further applications are imaginable ranging from the production of metabolites to bioremediation. Therefore, we would like to share our achievements with the iGEM community in the hope to support and facilitate future projects. Our supervisors Dr. Maximilian Ulbrich and Dr. Nicole Gensch provided us plasmids encoding the anti-GFP nanobody and the glutathione S-transferase that were inevitable for our project. The Integration vectors used to insert our constructs into the genome of B. subtilis were provided by Julia Bartels from the Mascher lab at the TU Dresden. Below you will find a table summarizing the submitted biobricks and a link guiding you to the entries in the registry.
|BBa_K2114998||pelB anti-GFP nanobody||pIG16_162|
2. Improved Biobricks
We submitted an improved anti-GFP nanobody containing the pelB leader sequence for the export of the protein in the periplasmatic space resulting in an increased yield of the over-expressed protein.
The B. subtilis promoter PCotYZ regulates the expression of the late stage spore coat proteins CotY and CotZ. We included a ribosome binding site which simplifies the assembly of devices and the expression of subsequent genes.