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Application

There are many possible applications for our Sporobeads as it is possible to modify their outermost protein layer. This way they could be used as beads with special functions. To easily create any kind of Sporobead we designed a Sporovector, where you just have to insert your gene of choice. Since there are so many possible applications we picked three examplary ideas for future Sporobeads:

Kumamolisin-Sporobeads: the solution for carefree enjoyment of everyday meals

World wide one out of [http://www.enriquecastro.net/index.php/term/,9da4ab975b545ba0ae53646c58a5a265aa5d535892a89b979fa4b1a49297a261a260555c5a.xhtml 3350] people cannot eat meals that contain wheat products and other foods with traces of gluten. Kumamolisin is an [http://partsregistry.org/wiki/index.php?title=Part:BBa_K590087 enzyme] that cleaves peptides and was produced by the iGEM-Team from the University of Washington] last year. The substrate includes a specific sequence of amino acids which causes celiac disease in sensitive people when they consume food containing gluten. Our beads could carry Kumamolisin and offer a protected passage through the stomach, so that the enzyme can work properly where it is needed in the intestines. The GerminationSTOP we put in place in our spores would ensure a correct dosage. This project is a pharmaceutical application and therefore would have to fulfill the laws for pharmaceuticals. This includes several verification steps of non-toxicity and efficacy.

CPX-Sporobeads: rescuing marine life

The excessive use of disposable plastic and the lack of universal recycling programs has led to the pollution of the world's oceans. In the ocean, large pieces of Polystyrene litter are ground by sea currents into very small pieces, so called plastik plankton, that are consumed by fish, filter feeders, and other organisms living in the oceans. Such plastic uptake can lead to poisoning, sterility and death. The [http://partsregistry.org/wiki/index.php/Part:BBa_I728500 CPX-peptide] generated by MIT (2007) can bind to Polystyrene. CPX-Sporobeads in huge filter boxes could be put into place to mechanically filter microscopic plastic particles, like polystyrene plankton, out of the water. To prevent the beads from being released into the sea and the plastic to be kept the Sporobeads could be attached to membranes in the boxes. The spores would then not only express CPX but also a membrane binding protein on their surface.

TALE-Sporobeads: easy and cheap detection of GMOs

Since 1990, green biotechnology releases many transgenic plants into the environment by selling genetically modified seeds. Thus organic farmers need to prove today, that their products meet the requirements for organic crops. Usually they pay laboratories to attest the lack of contamination with genetically modified crops. The new tools of molecular biology, TAL effectors, combined with our Sporobeads could be an easy and cheap solution for organic agriculture. Farmers could in future use our kit with TALE-lacZ-Sporobeads to detect genetically modified crops themselves. As spores are stable and safe vehicles, they could be send by mail without any considerations. The kit would be suitable for use outside of laboraty. The protocol for this could work as follows:
The DNA extracted from plants with solutions provided by the kit, is immobilized and fixed on a nitrocellulose membrane. This membrane is then washed, incubated with Sporobeads in solution and washed again. With addition of the substrate X-Gal, the lacZ of bound Sporobeads will catalyze the reaction so that a blue staining appears. If no such DNA is present, the spores will not bind and no blue color will appear.

In general, B. subtilis spores are suitable for high-scale applications: they are

• easy and cheap to produce in large quantities ([http://www.alibaba.com/product-free/118791994/Bacillus_subtilis_spores_HU58_100_pure.html offer])
• very stable in several extreme environment conditions
• unable to proliferate and therefore most likely not treated as genetically modified organisms by law
• part of human nutrition in [http://en.wikipedia.org/wiki/Bacillus_subtilis_R0179 food supplements] and generally regarded as safe

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