Team:LMU-Munich/Project

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== '''Overall project''' ==
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<html>
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[[File:Project draft.png|500px|right]]
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<style type="text/css">
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The goal of our project BEADzillus is the production of ''Bacillus subtilis'' spores which display different proteins with special features on their surface.
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#buttons {
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For example, the spore could:
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position:relative;
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}
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- bind harmful viruses
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#buttons img {
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display: block;
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position: absolute;
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top: 0;
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left: 0;
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z-index: -100;
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}
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- bind toxic metals
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#buttons a img {
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display: none;
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}
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- bind plastic molecules
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#buttons a:hover img {
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display: block;
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}
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- expose enzymes
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#buttons a {
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display: block;
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width: 50px;
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height: 50px;
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position: absolute;
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z-index 100;
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/* outline: 1px solid red; */
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}
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</style>
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<div id="buttons" style="height: 375px;">
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<img alt="" src="http://wrtlprnft.ath.cx/igem2012_home.png">
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<a href="https://2012.igem.org/Team:LMU-Munich/Project" style="left: 276px; top: 40px; width: 271px; height:265px;"><img style="left: -276px; top: -40px" alt="" src="http://wrtlprnft.ath.cx/igem2012_home0.png"></a>
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<a href="https://2012.igem.org/Team:LMU-Munich/Spore_Coat_Proteins" style="left: 130px; top: 50px; width: 146px; height:145px;"><img style="left: -130px; top: -50px" alt="" src="http://wrtlprnft.ath.cx/igem2012_home1.png"></a>
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<a href="https://2012.igem.org/Team:LMU-Munich/Germination_Stop" style="left: 166px; top: 196px; width: 110px; height:110px;"><img style="left: -166px; top: -196px" alt="" src="http://wrtlprnft.ath.cx/igem2012_home2.png"></a>
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<a href="https://2012.igem.org/Team:LMU-Munich/Bacillus_BioBricks" style="left: 55px; top: 160px; width: 110px; height:110px;"><img style="left: -55px; top: -160px" alt="" src="http://wrtlprnft.ath.cx/igem2012_home3.png"></a>
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</div>
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</html>
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to filter fluids  or for use in laboratory.
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<p align=justify>
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Our project '''Bead'''zillus is composed of several modules. Click on the symbols to learn more about their function.
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As a safety issue we will stop spores from outgrowing. First we want to knock out several genes which are important for germination and second we want potentially outgrowing spores to be killed by a toxin they produce themselves.
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The idea of '''Bead'''zillus is to enrich the [http://partsregistry.org/Main_Page partsregistry] with new BioBricks that are needed for the work with ''B. subtilis'' and show how its unique features lead to new creative ideas. We use the spores of ''B. subitlis'' to establish a platform for protein display.
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</p>
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== Project Details==
 
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[[File:Project arrow 1.png|500px|center]]
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*[[File:Bacilluss_Intro.png|30px|link=Team:LMU-Munich/Bacillus_Introduction]] [[Team:LMU-Munich/Bacillus_Introduction|'''''Bacillus'' Intro''']]: A short introduction to the organism ''Bacillus subtilis'' and why we chose to work with it.
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*[[File:BacillusBioBrickBox.png|30px|link=Team:LMU-Munich/Bacillus_BioBricks]] Our [[Team:LMU-Munich/Bacillus_BioBricks|'''''Bacillus''B'''io'''B'''rick'''B'''ox]] offers a set of new parts for the work with ''B. subtilis''.
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*[[File:SporeCoat.png|30px|link=Team:LMU-Munich/Spore_Coat_Proteins]] [[Team:LMU-Munich/Spore_Coat_Proteins|'''Sporo'''beads]] are a platform to display proteins on ''B. subtilis'' spores.
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*[[File:GerminationSTOP.png|25px|link=Team:LMU-Munich/Germination_Stop]] [[Team:LMU-Munich/Germination_Stop|'''Germination'''STOP]] prevents the spores from germinating to vegetative cells.
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*On our [[Team:LMU-Munich/Data|Data]] page you find all our results in detail.
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*Why did we chose spores to display proteins and for what could they be used? See our [[Team:LMU-Munich/Application|Applications page]].
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=== Clone Vectors ===
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Part of establishing ''Bacillus subtilis'' to the BioBrick mode is to modify standard vectors of ''Bacillus''. We mutate prohibited restriction sites (EcoRI, XbaI, SpeI, PstI) and excise redundant parts of the vectors.  
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Visit the other pages related to our project:
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* [[Team:LMU-Munich/Attributions|Attributions]] who helped us.
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* [[Team:LMU-Munich/Collaborations|Collaborations]] with other iGEM Teams.
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* [[Team:LMU-Munich/Parts|Registry]] Here is the list of all new BioBricks we've added to the registry so far.
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* [[Team:LMU-Munich/Strains|''Bacillus'' strains]] Here you find information about the ''B. subtilis'' strains we created and how to obtain them.
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* [[Team:LMU-Munich/Inverter|Inverter]] See here a special project we continued from the LMU-Munich 2011 Team.
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* [[Team:LMU-Munich/Achievements|Achievements]] Find here which medal we would give ourselves and why.
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=== Evaluate Promoters ===
 
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Another point is to produce promoters of Bacillus subtilis in BioBrick mode and to evaluate them. We amplify them of the Bacillus genome and clone them upstream of reporter genes (GFP, lux operon, lacZ) to measure their activity in ''Bacillus subtilis''.
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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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=== Fuse Spore Coat Proteins ===
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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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There are several different coat proteins in the spores of Bacillus subtilis and we chose CotZ and CgeA which are located in the outermost layer. The first step is to  fuse GFP to these proteins to see if they appear on the surface and if there is any affection of spore formation. The next step is to fuse proteins with special features to CotZ and CgeA to produce functional SporoBeads.
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|[[File:SporeCoat.png|100px|link=Team:LMU-Munich/Spore_Coat_Proteins]]
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These properties could be:
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|[[File:GerminationSTOP.png|100px|link=Team:LMU-Munich/Germination_Stop]]
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- binding harmful viruses
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|-
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- binding toxic metals
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|[[Team:LMU-Munich/Bacillus_Introduction|<font size="2">'''''Bacillus'''''<BR>Intro</font>]]
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- binding plastic molecules
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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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- expose enzymes
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|[[Team:LMU-Munich/Spore_Coat_Proteins|<font size="2" face="verdana">'''Sporo'''beads</font>]]
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to filter fluids or for use in the laboratory.
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|[[Team:LMU-Munich/Germination_Stop|<font size="2" face="verdana">'''Germination'''<BR>STOP</font>]]
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|}
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</div>
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=== Stop Germination ===
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To stop the „SporoBeads“ from germination, we thought of two different manipulations. First, we knock-out several genes which are necessary for all germination pathways, namely the cortex hydrolysis. Several gene deletion combinations will be tested for their outgrowth rates.
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As a second mechanism, spores which begin to germinate despite the gene deletions shall be killed by an endotoxin system. Here, we work with a sigma-factor from a different species which works in ''Bacillus subtilis'' but does not interfere with endogenous promoters. This sigma factor is placed downstream of a promoter that is strongly activated by sigmaG , the last active sigma factor in the endospore. Therefore, the alternative sigma factor is produced right before sporulation. The bacteria also have a second construct inserted into their genome: a promoter activated by the alternative sigma factor which leads to the transcription of an endotoxin. So, if a spore starts metabolism, the toxin gene will be activated and kill the outgrowing spore.
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== Results ==
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{{:Team:LMU-Munich/Templates/Page Footer}}
{{:Team:LMU-Munich/Templates/Page Footer}}

Latest revision as of 12:25, 26 October 2012

iGEM Ludwig-Maximilians-Universität München Beadzillus

Team-LMU red and white.resized.jpg

The LMU-Munich team is exuberantly happy about the great success at the World Championship Jamboree in Boston. Our project Beadzillus finished 4th and won the prize for the "Best Wiki" (with Slovenia) and "Best New Application Project".

IGEM HQ LMU prize.jpg

[ more news ]

Sporenfreunde

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