Team:LMU-Munich/Spore Coat Proteins

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==Spore Coat Proteins==
 
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The aim of this project part 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 (Imamura et al., 2011).This is why we used them to create functional fusion proteins to be expressed on our '''Sporo'''beads.
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[[File:SporeCoat.png|100px|right|link=]]
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The gene ''cgeA'' is located in the ''cgeABCDE''cluster and is regulated by its own promoter P<sub>''cgeA''</sub>. The cluster ''cotVWXYZ'' contains the gene ''cotZ'' which is cotranscribed with ''cotY'' regulated by the promoter P<sub>''cotYZ''</sub>.
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Our first step will be to  fuse GFP to these proteins to see if they appear on the spore surface and if there is any effect on spore formation. The next step will be to fuse proteins with special features to CotZ and CgeA to produce functional "SporoBeads."
 
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SporoBeads could be used to filter fluids or in the laboratory, and could be capable of:
 
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- binding harmful viruses
 
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- binding toxic metals
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=='''Sporo'''beads - What Protein Do ''You'' Want to Display?==
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<br>
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<p align="justify">'''Sporo'''beads are the first generation of ''Bacillus subtilis'' endospores displaying a protein of our choice on their outermost layer, the spore crust. In future '''Sporo'''beads could serve as a platform for protein display and thus be used for numerous versatile [https://2012.igem.org/Team:LMU-Munich/Application applications]. Our goal was to show that ''B. subtilis'' spores have the ability to do so. As a [https://2012.igem.org/Team:LMU-Munich/Spore_Coat_Proteins/result proof of principle] we successfully fused GFP to the spore crust and obtained fluorescence with microscopy.</p> 
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- binding plastic molecules
 
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- exposing enzymes
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<div class="box">
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===Scientific Background===
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{| "width=100%" style="text-align:center;" style="align:right"|
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|<p align="justify">Introduction to ''B. subtilis'' spores  and their use in our project</p>
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|[[File:Imamura, 2011 &amp; McKenney, 2010.png|200px|right|link=Team:LMU-Munich/Spore_Coat_Proteins/Background]]
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! colspan="2" |[[File:LMU Arrow purple.png|40px|link=Team:LMU-Munich/Spore_Coat_Proteins/Background]]
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===Cloning Strategy===
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{| "width=100%" style="text-align:center;" style="align:right"|
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|<p align="justify">Cloning strategy to create different variants of our ''' Sporo'''beads</p>
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|[[File:Final construct.png|200px|link=Team:LMU-Munich/Spore_Coat_Proteins/cloning]]
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! colspan="2" |[[File:LMU Arrow purple.png|40px|link=Team:LMU-Munich/Spore_Coat_Proteins/cloning]]
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|}
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'''Project Navigation'''
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<div class="box">
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===GFP as a Proof of Principle===
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{| "width=100%" style="text-align:center;" style="align:right"|
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|<p align="justify">Main results of the various constructs that were created to find the best one!</p>
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|[[File:LMU Firstspore.jpg|200px|right|link=Team:LMU-Munich/Spore_Coat_Proteins/result]]
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! colspan="2" |[[File:LMU Arrow purple.png|40px|link=Team:LMU-Munich/Spore_Coat_Proteins/result]]
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|}
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</div>
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{|
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|<font color="#FFFFFF">+--+--+--+--+--+--+--</font>
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===Laccases as functional enzymes===
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|<font color="#FFFFFF">+--+--+--+--+--</font>
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{| "width=100%" style="text-align:center;" style="align:right"|
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|<font color="#FFFFFF">+--+--+--+--+--+--</font>
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|<p align="justify">Creation of functional Laccase-''' Sporo'''beads</p>
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|<font color="#FFFFFF">+--+--+--+--+--+--</font>
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|[[File:Final construct.png|200px|link=Team:LMU-Munich/Spore_Coat_Proteins/laccases]]
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! colspan="2" |[[File:LMU Arrow purple.png|40px|link=Team:LMU-Munich/Spore_Coat_Proteins/laccases]]
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|}
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<a href="https://2012.igem.org/Team:LMU-Munich/Bacillus_Introduction">
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<img src="https://static.igem.org/mediawiki/2012/8/8d/Bacilluss_Intro.png" height=70"/>
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<a href="https://2012.igem.org/Team:LMU-Munich/Bacillus_Introduction"><font size="1" face="verdana">Bacillus<BR>Intro</font></a>
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|<html>
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<a href="https://2012.igem.org/Team:LMU-Munich/Bacillus_BioBricks">
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===Purification Methods===
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<img src="https://static.igem.org/mediawiki/2012/7/78/BacillusBioBrickBox.png" height=70"/>
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{| "width=100%" style="text-align:center;" style="align:right"|
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</a>
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|<p align="justify">Description of the different purification methods of the spores</p>
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|[[File:Treatments.png|200px|right|link=Team:LMU-Munich/Spore_Coat_Proteins/purification]]
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! colspan="2" |[[File:LMU Arrow purple.png|40px|link=Team:LMU-Munich/Spore_Coat_Proteins/purification]]
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<a href="https://2012.igem.org/Team:LMU-Munich/Bacillus_BioBricks"><font size="1" face="verdana">Bacillus<BR>BioBrickBOX</font></a>
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|}
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<div class="box">
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====Project Navigation====
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{| width="100%" align="center" style="text-align:center;"
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|[[File:Bacilluss_Intro.png|100px|link=Team:LMU-Munich/Bacillus_Introduction]]
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|[[File:BacillusBioBrickBox.png|100px|link=Team:LMU-Munich/Bacillus_BioBricks]]
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|[[File:SporeCoat.png|100px|link=Team:LMU-Munich/Spore_Coat_Proteins]]
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|[[File:GerminationSTOP.png|100px|link=Team:LMU-Munich/Germination_Stop]]
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|[[Team:LMU-Munich/Bacillus_Introduction|<font size="2">'''''Bacillus'''''<BR>Intro</font>]]
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|[[Team:LMU-Munich/Bacillus_BioBricks|<font size="2" face="verdana">'''''Bacillus'''''<BR>'''B'''io'''B'''rick'''B'''ox</font>]]
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|[[Team:LMU-Munich/Spore_Coat_Proteins|<font size="2" face="verdana">'''Sporo'''beads</font>]]
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|[[Team:LMU-Munich/Germination_Stop|<font size="2" face="verdana">'''Germination'''<BR>STOP</font>]]
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|<html>
 
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<a href="https://2012.igem.org/Team:LMU-Munich/Spore_Coat_Proteins">
 
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<a href="https://2012.igem.org/Team:LMU-Munich/Spore_Coat_Proteins"><font size="1" face="verdana">SporeCoat<BR>FusionProteins</font></a>
 
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<a href="https://2012.igem.org/Team:LMU-Munich/Germination_Stop">
 
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<a href="https://2012.igem.org/Team:LMU-Munich/Germination_Stop"><font size="1" face="verdana">Germination<BR>STOP</font></a>
 
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Latest revision as of 13:44, 14 November 2012

iGEM Ludwig-Maximilians-Universität München Beadzillus

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