Team:LMU-Munich/Spore Coat Proteins

From 2012.igem.org

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==Spore Crust Proteins==  
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=='''Sporo'''beads - what protein do you want them to display?==  
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<p align="justify">The aim of this module is to create spores that display fusion proteins on their crust. There are several different proteins forming the spore coat layers of ''Bacillus subtilis'' spores. On the outermost layer, the so called spore crust, the CotZ and CgeA proteins are located ([http://www.ncbi.nlm.nih.gov/pubmed?term=imamura%20et%20al.%202011%20spore%20crust Imamura et al., 2011]). This is why we used them to create functional fusion proteins to be expressed on our '''Sporo'''beads.</p>  
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<p align="justify">'''Sporo'''beads are ''Bacillus subtilis spores modulated in a way that they can be used as a platform for individual protein display. The aim of this module is to create these spores that display fusion proteins on their'' crust. There are several different proteins forming the spore coat layers of ''Bacillus subtilis'' spores. On the outermost layer, the so called spore crust, the CotZ and CgeA proteins are located ([http://www.ncbi.nlm.nih.gov/pubmed?term=imamura%20et%20al.%202011%20spore%20crust Imamura et al., 2011]). This is why we used them to create functional fusion proteins to be expressed on our '''Sporo'''beads.</p>  
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==Applications==
==Applications==
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<p align="justify">There are many possible applications for our '''Sporo'''beads and to easily create any kind of spore we designed a [https://2012.igem.org/Team:LMU-Munich/Bacillus_BioBricks/Sporovector '''Sporo'''vector]. With the help of this vector one can produce any individually desired spore. We picked three exemplary applications for our '''Sporo'''beads to be explained in detail: </p>
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<p align="justify">There are many possible applications for our '''Sporo'''beads 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 '''Sporo'''bead we designed a [https://2012.igem.org/Team:LMU-Munich/Bacillus_BioBricks/Sporovector '''Sporo'''vector], where you just have to insert your protein of choice. Since there are so many possible applications we picked three examplary ideas for future '''Sporo'''beads: </p>
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<br>'''Kumamolisin:'''  
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<br>'''Kumamolisin:'''- the solution for carefree enjoyment of everyday meals
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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 [https://2011.igem.org/Team:Washington 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 this enzyme and offer a protected passage through the stomach, so that the enzyme can work where it is needed in the intestines. The <b>Germination</b>STOP 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.
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World wide one out of [http://www.enriquecastro.net/index.php/term/,9da4ab975b545ba0ae53646c58a5a265aa5d535892a89b979fa4b1a49297a261a260555c5a.xhtml 3350] people cannot eat that contains 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 [https://2011.igem.org/Team:Washington 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 [https://2012.igem.org/Team:LMU-Munich/Germination_Stop <b>Germination</b>STOP] 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.
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'''CPX:'''  
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'''CPX:''' - the relieve of the marine life
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The [http://partsregistry.org/wiki/index.php/Part:BBa_I728500 CPX-peptide] generated by MIT (2007) can bind to Polystyrene which is a common plastic. The waste from Polystyrene is currently a big challenge in the field of environmental protection and remediation. 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 leads to poisoning, sterility and death. To remove Polystyrene from the oceans, huge filter boxes could be put into place to mechanically filter out large pieces; our CPX-Sporobeads could remove the microscopic plastic particles.  
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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-'''Sporo'''beads 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 '''Sporo'''beads 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.
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Accidentally, some spores could loosen from the surface they would be affixed to and float into the sea. Therefore, we would have to ask the ZKBS([http://www.bvl.bund.de/DE/06_Gentechnik/03_Antragsteller/06_Institutionen_fuer_biologische_Sicherheit/01_ZKBS/gentechnik_zkbs_node.html Zentrale Kommission für biologische Sicherheit]; “Central Committee for Biological Safety”) for permission to release of the spores. They would require details about our spores and check whether our spores would be safe for environmental release.
 
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Another application could be the removal of plastic particles in paper recycling facilities. The applicable laws and regulations should be similar.
 
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'''Virus binding:'''  
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'''TALEs''' - the
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Viruses bind specifically to certain proteins and domains to infect cells. To capitalize on this mechanism, the <b>Sporo</b>beads could be coated with such domains. The viruses would then bind to the spores and could be removed from liquid samples. This could be used to filter human immunodeficiency virus (HIV) from blood for transfusions, or even to help people with deadly virus infections by dialyzing their blood and thereby lowering their virus titer.
 
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Of course, permission from the ZKBS would be necessary, but because virus binding would be a medical application, rules from the medical field must be taken into account. The Robert-Koch-Institute in charge of hospital hygiene and infection prevention would check for the hygienic methods of such an application and whether it would work according to the [http://www.gesetze-im-internet.de/ifsg/ Infektionsschutzgesetz] (“prevention of infections law”).
 
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Consideration of medical laws would also be necessary, if our <b>Sporo</b>beads were to be used to filter viruses from drinking water.
 

Revision as of 17:21, 26 September 2012

iGEM Ludwig-Maximilians-Universität München Beadzillus

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