Team:LMU-Munich/Spore Coat ProteinsTour/Background

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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 surface. There are several different proteins forming the spore coat layers of ''B. subtilis'' spores (Fig. 1). The outermost layer, the so-called spore crust, is composed of two proteins, CotZ and CgeA ([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 modified so 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 surface. There are several different proteins forming the spore coat layers of ''B. subtilis'' spores (Fig. 1). The outermost layer, the so-called spore crust, is composed of two proteins, CotZ and CgeA ([http://www.ncbi.nlm.nih.gov/pubmed?term=imamura%20et%20al.%202011%20spore%20crust Imamura et al., 2011]). This is why we used these two crust proteins to create functional fusion proteins to be expressed on our '''Sporo'''beads.</p>  
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|[[File:Imamura, 2011 &amp; McKenney, 2010.png|Protein distribution in spore coat of ''Bacillus subtilis''|500px|center]]
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=====Crust Protein Genes Organization=====
=====Crust Protein Genes Organization=====
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<p align="justify">The gene ''cgeA'' is located in the ''cgeABCDE'' cluster and is expressed from its own promoter P<sub>''cgeA''</sub>. The cluster ''cotVWXYZ'' contains the gene ''cotZ'' which is cotranscribed with ''cotY'' and expressed from the promoter P<sub>''cotYZ''</sub>. Another promoter of this cluster, P<sub>''cotV''</sub>, is responsible for the transcription of the other three genes (Fig. 2). Those three promoters were evaluated with the ''lux'' reporter genes ([http://partsregistry.org/Part:BBa_K823025 pSB<sub>''Bs''</sub>3C-''lux''ABCDE]) to analyze their strength and the time point of their activation (see [https://2012.igem.org/Team:LMU-Munich/Data/crustpromoters data]).</p>  
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<p align="justify">The gene ''cgeA'' is located in the ''cgeABCDE'' cluster and is expressed from its own promoter P<sub>''cgeA''</sub>. The cluster ''cotVWXYZ'' contains the gene ''cotZ'' which is cotranscribed with ''cotY'' and expressed from the promoter P<sub>''cotYZ''</sub>. Another promoter of this cluster, P<sub>''cotV''</sub>, is responsible for the transcription of the other three genes (Fig. 2). Those three promoters were evaluated with the ''lux'' reporter genes (pSB<sub>''Bs''</sub>3C-''lux''ABCDE) to analyze their strength and the time point of their activation (visit later DATA page). Based on this data, two of the three promoters could be used for expression of spore crust fusion proteins.
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Based on this data, two of the three promoters could be used for expression of spore crust fusion proteins.
 

Latest revision as of 16:46, 26 October 2012

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