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content.i = 2;

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content.childs=[3,11,9,10,7];

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content.childs=[4,5,9,11,7,10];

content.titleShort = "Protein regulation";

content.titleLong = "Protein regulation mechanisms";

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content.parents=[2]

content.childs=[];

content.titleShort = "Protein cleavage";

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content.parents=[2];

content.childs=[];

content.titleShort = "Gene knock-out and knock-in";

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content.titleShort = "Direct inhibition and activation";

content.titleLong = "Direct inhibition and activation";

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content.summary= "Direct binding of small molecules an activate or inactivate protein function";

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content.summary= "Direct binding of small molecules in order to activate or inactivate protein function";

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content.text= "Moreover proteins can be regulated directly in many different ways. Binding of other Proteins or small molecules for example can activate or deactivate the catalytic function of proteins. For example small molecules can have great impact on protein function. They can often influence protein functions by binding to an allosteric center. Small molecules can be activating, as well as repressing, depending on the type of the molecule<sup><a href=#11.1>11.1</a></sup>.</br></br><div class='content-image'> <img src=https://static.igem.org/mediawiki/2013/1/1c/BonnSmallMolecules.jpg></br>Different mechanisms for small-molecule activation of enzymes<sup><a href=#11.2>11.2</a></sup> </div></br></br>The following picture shows the example of the allosteric activation of a glucokinase</br> <div class='content-image'> <img src=https://static.igem.org/mediawiki/2013/f/f4/BonnSmallMolecules2.jpg></br>"(a) GK bound to the GKA, compound A, and glucose (blue, Protein Data Bank (PDB) ID 1V4S). Compound A binds at a site distal from the active site, which is highlighted by the presence of the substrate, glucose. (b) Structural overlay of GK in the presence of compound A and glucose with an unliganded, inactive GK (pink, PDB ID 1V4T). In the unbound GK, the GKA binding site is occluded. A large shift in the small subunit of GK occurs from the unbound to bound structures (black arrow). Glucose promotes the active conformation, which is hindered from shifting back to the inactive conformation in the presence of compound A. (c) 7 mutations (out of 13)9 identified in GK (pink) that are associated with disease map to the GKA binding site. These mutations highlight an important regulatory site within GK, and could similarly stabilize a closed, active conformation."<sup><a href=#11.2>[11.2]</a></sup></div></br></br><p><a name=11.1>11.1</a><a href='http://www.nature.com/nrd/journal/v3/n4/full/nrd1343.html'>Small-molecule inhibitors of proteinprotein interactions: progressing towards the dream; Michelle R. Arkin & James A. Wells</a>/p></br><p><a name=11.2>11.2</a><a href='http://www.nature.com/nchembio/journal/v6/n3/full/nchembio.318.html'>Turning enzymes ON with small molecules, Julie A Zorn & James A Wells</a></p>";

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content.text= "Direct binding of small molecules an activate or inactivate protein function";

+

content.text= "Moreover proteins can be regulated directly in many different ways. Binding of other Proteins or small molecules for example can activate or deactivate the catalytic function of proteins. For example small molecules can have great impact on protein function. They can often influence protein functions by binding to an allosteric center. Small molecules can be activating, as well as repressing, depending on the type of the molecule<sup><a href=#11.1>11.1</a></sup>.</br></br><div class='content-image'> <img src='https://static.igem.org/mediawiki/2013/1/1c/BonnSmallMolecules.jpg' width='550'> </br>Different mechanisms for small-molecule activation of enzymes<sup><a href=#11.2>11.2</a></sup> </div></br></br>The following picture shows the example of the allosteric activation of a glucokinase</br> <div class='content-image'> <img src=https://static.igem.org/mediawiki/2013/f/f4/BonnSmallMolecules2.jpg width='600'></br>"(a) GK bound to the GKA, compound A, and glucose (blue, Protein Data Bank (PDB) ID 1V4S). Compound A binds at a site distal from the active site, which is highlighted by the presence of the substrate, glucose. (b) Structural overlay of GK in the presence of compound A and glucose with an unliganded, inactive GK (pink, PDB ID 1V4T). In the unbound GK, the GKA binding site is occluded. A large shift in the small subunit of GK occurs from the unbound to bound structures (black arrow). Glucose promotes the active conformation, which is hindered from shifting back to the inactive conformation in the presence of compound A. (c) 7 mutations (out of 13)9 identified in GK (pink) that are associated with disease map to the GKA binding site. These mutations highlight an important regulatory site within GK, and could similarly stabilize a closed, active conformation."<sup><a href=#11.2>[11.2]</a></sup></div></br></br><p><a name=11.1>11.1</a><a href='http://www.nature.com/nrd/journal/v3/n4/full/nrd1343.html'>Small-molecule inhibitors of proteinprotein interactions: progressing towards the dream; Michelle R. Arkin & James A. Wells</a>/p></br><p><a name=11.2>11.2</a><a href='http://www.nature.com/nchembio/journal/v6/n3/full/nchembio.318.html'>Turning enzymes ON with small molecules, Julie A Zorn & James A Wells</a></p>";

content.type="Background";

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content.titleLong = "Ec. SspB adaptor";

content.summary= "The sspB protein is an adaptor responsible for delivering ssrA-tagged substrates to the ClpXP protease in order to enhance their degradation";

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content.text= "<b>Introduction </b> </br> The sspB protein is an adaptor responsible for delivering ssrA-tagged substrates to the ClpXP protease in order to enhance their degradation.Thus, bacterias like E.coli or C.crescentus regulate the concentration of marked proteins and also are in control of their quality. Even though degeneration of tagged substrates is possible without sspB, sspB delivering is a common process, because it improves the affinity between ssrA and ClpXP<sup><a href='#14.1'>14.1</a></sup>. </br> In our project, we used a sspB split variant instead of the normal sspB in order to control cleaving rate. The two parts of this version only stay divided until we ray them with light of a special wavelength. After that, the fractions form an unit and can function normal from now on as described below. </br> </br><b>Structure </b> </br>The sspB adaptor is a homomeric dimer (Fig. 1), which means that itconsists of two identical domains. Together the domains form a pore with the ssrA binding sites inside. Each domain also owns a C-terminal tail ending in a ClpX binding module, called XP. The amino-acid sequence of XP is highly conserved so that mutations in it mostly cause extremely decrease in activity, whereas the linker sequence differs from species to species <sup><a href='#14.2'>14.2</a></sup><sup><a href='#14.3'>14.3</a></sup>. <div align='left'><img src='https://static.igem.org/mediawiki/2013/5/5c/BonnsspB_Fig2.jpg' height='202' width='403'>Fig. 1: Ribbon diagramm of the sspB dimer with a bound srrA-tagged protein, from 'Versatile modes of peptide recognition by the AAA+ adaptor protein SspB, Levchenko et al, 2005, nature structural and molecular biology, PMID: 15880122</div> </br> </br></br></br><b>Function</b></br> SspB enhances degradation of ssrA-tagged proteins by lowering the K<sub>M</sub>. Thus, with a given substrate concentration sspB-mediated cleaving runs faster than without sspB (Fig. 2) </br> <div align='left'><img src='https://static.igem.org/mediawiki/2013/d/d8/BonnSspB_Fig1.jpg' height='232' width='371'>Fig. 2: Diagramm, shows the degradation rate of GFP-ssrA without sspB, with sspB and with two mutations, the given substrate concentration is 0.3 &my;M, from 'Flexible Linkers Leash the Substrate Binding Domain of SspB to a Peptide Module that Stabilizes Delivery Complexes with the AAA ClpXP Protease, Wah et al, Molecular Cell, 2003, PMID: 14536075' </div></br> Therefor, the sspB dimer contains a pore and while 'AADENY' is linked with the inside, the 'LAA'-domain (respectively 'DAS') faces outwards, free to bind ClpX (Fig. 3).</br> The affinity of this binding amounts around 20 &my;M, which suggests a relative strong connection. The two extremely flexible ClpX binding tails with XP at the C-terminal end dock on ClpX. So the 'LAA'- domain lies closely to ClpX's axial pore and can be bound to it. <div align='left'><img src='https://static.igem.org/mediawiki/2013/7/7c/BonnSsra_fig3.jpg' height='151' width='116'>Fig. 3: Model of sspB with a bound ssrA-tagged substrate, from 'Engineering controllable protein degradation, McGinness et al, Molecular cell, 2006, PMID:16762842' </div></br> To sum up, there are three bonds connecting the ssrA-sspB-ClpX-complex and making it relative stable: ssrA with sspB, sspB with ClpX and ssrA with ClpX. Hence follows a lower K<sub>M</sub> than the direct binding process has<sup><a href='#14.4'>14.4</a></sup><sup><a href='#14.5'>14.5</a></sup>. <h2> References </h2> <a id='14.1'>14.1</a>Bivalent Tethering of SspB to ClpXP Is Required for Efficient Substrate Delivery: A Protein-Design Study, Bolon et al, 2004, Molecular Cell, PMID: 14967151 </br><a id='14.2'>14.2</a>see above </br> <a id='14.3'>14.3</a> Flexible Linkers Leash the Substrate Binding Domain of SspB to a Peptide Module that Stabilizes Delivery Complexes with the AAA ClpXP Protease, Wah et al, Molecular Cell, 2003, PMID: 14536075 </br> <a id='14.4'>14.4</a> see above</br> <a id='14.6'>[14.5]</a> Altered Tethering of the SspB Adaptor to the ClpXP Protease Causes Changes in Substrate Delivery, McGinnes KE et al, The journal of Biological Chemistry, 2007, PMID: 17317664 </br> <a id='14.4'>14.4</a> ClpXP, an ATP-powered unfolding and protein-degradation machine, Baker et al, Biochim Biophys Acta, 2012, PMCID: PMC3209554</br>";

+

content.text= "<b>Introduction </b> </br> The sspB protein is an adaptor responsible for delivering ssrA-tagged substrates to the ClpXP protease in order to enhance their degradation.Thus, bacterias like E.coli or C. crescentus regulate the concentration of marked proteins and also are in control of their quality. Even though degeneration of tagged substrates is possible without sspB, sspB delivering is a common process, because it improves the affinity between ssrA and ClpXP<sup><a href='#14.1'>14.1</a></sup>. </br> In our project, we used a sspB split variant instead of the normal sspB in order to control cleaving rate. The two parts of this version only stay divided until we ray them with light of a special wavelength. After that, the fractions form an unit and can function normal from now on as described below. </br> </br><b>Structure </b> </br>The sspB adaptor is a homomeric dimer (Fig. 1), which means that itconsists of two identical domains. Together the domains form a pore with the ssrA binding sites inside. Each domain also owns a C-terminal tail ending in a ClpX binding module, called XP. The amino-acid sequence of XP is highly conserved so that mutations in it mostly cause extremely decrease in activity, whereas the linker sequence differs from species to species <sup><a href='#14.2'>14.2</a></sup><sup><a href='#14.3'>14.3</a></sup>. <div align='left'><img src='https://static.igem.org/mediawiki/2013/5/5c/BonnsspB_Fig2.jpg' height='202' width='403'>Fig. 1: Ribbon diagramm of the sspB dimer with a bound srrA-tagged protein, from 'Versatile modes of peptide recognition by the AAA+ adaptor protein SspB, Levchenko et al, 2005, nature structural and molecular biology, PMID: 15880122</div> </br> </br></br></br><b>Function</b></br> SspB enhances degradation of ssrA-tagged proteins by lowering the K<sub>M</sub>. Thus, with a given substrate concentration sspB-mediated cleaving runs faster than without sspB (Fig. 2) </br> <div align='left'><img src='https://static.igem.org/mediawiki/2013/d/d8/BonnSspB_Fig1.jpg' height='232' width='371'>Fig. 2: Diagramm, shows the degradation rate of GFP-ssrA without sspB, with sspB and with two mutations, the given substrate concentration is 0.3 &my;M, from 'Flexible Linkers Leash the Substrate Binding Domain of SspB to a Peptide Module that Stabilizes Delivery Complexes with the AAA ClpXP Protease, Wah et al, Molecular Cell, 2003, PMID: 14536075' </div></br> Therefor, the sspB dimer contains a pore and while 'AADENY' is linked with the inside, the 'LAA'-domain (respectively 'DAS') faces outwards, free to bind ClpX (Fig. 3).</br> The affinity of this binding amounts around 20 &my;M, which suggests a relative strong connection. The two extremely flexible ClpX binding tails with XP at the C-terminal end dock on ClpX. So the 'LAA'- domain lies closely to ClpX's axial pore and can be bound to it. <div align='left'><img src='https://static.igem.org/mediawiki/2013/7/7c/BonnSsra_fig3.jpg' height='151' width='116'>Fig. 3: Model of sspB with a bound ssrA-tagged substrate, from 'Engineering controllable protein degradation, McGinness et al, Molecular cell, 2006, PMID:16762842' </div></br> To sum up, there are three bonds connecting the ssrA-sspB-ClpX-complex and making it relative stable: ssrA with sspB, sspB with ClpX and ssrA with ClpX. Hence follows a lower K<sub>M</sub> than the direct binding process has<sup><a href='#14.4'>14.4</a></sup><sup><a href='#14.5'>14.5</a></sup>. <h2> References </h2> <a id='14.1'>14.1</a>Bivalent Tethering of SspB to ClpXP Is Required for Efficient Substrate Delivery: A Protein-Design Study, Bolon et al, 2004, Molecular Cell, PMID: 14967151 </br><a id='14.2'>14.2</a>see above </br> <a id='14.3'>14.3</a> Flexible Linkers Leash the Substrate Binding Domain of SspB to a Peptide Module that Stabilizes Delivery Complexes with the AAA ClpXP Protease, Wah et al, Molecular Cell, 2003, PMID: 14536075 </br> <a id='14.4'>14.4</a> see above</br> <a id='14.6'>[14.5]</a> Altered Tethering of the SspB Adaptor to the ClpXP Protease Causes Changes in Substrate Delivery, McGinnes KE et al, The journal of Biological Chemistry, 2007, PMID: 17317664 </br> <a id='14.4'>14.4</a> ClpXP, an ATP-powered unfolding and protein-degradation machine, Baker et al, Biochim Biophys Acta, 2012, PMCID: PMC3209554</br>";

content.type="Background";

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content.i = 18;

content.parents=[17];

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content.childs=[19,21,20];

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content.childs=[36,21,20];

content.titleShort = "Light";

content.titleLong = "Light as a means of induction";

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case 21:

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content.i = 36;

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content.parents=[19];

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content.parents=[18];

content.childs=[37];

content.titleShort = "LOV-ipaA & VinD1";

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content.summary= "The LOV-ipaA -vinculin system is a combined system for light inducible heterodimerisation. This powerful tool, which allows photocontroled complex formation was establish by Lungu et al. in 2012.";

content.text= "The LOV-ipaA -vinculin system is a combined system for light inducible heterodimerisation. It consists out of a LOV domain, which undergoes conformational changes upon irradiation with blue light, and the ipaA-vinculin hybridization system. This two building blocks have be combined and described by Lungu et al. in 2012.</br></br> Lungu et al. (2008) where able to measure a 49-fold difference in target binding upon irradiation as compared to the dark state. However, they further modified the system by mutations of the LOV-ipaA construct and successfully weakend the baseline affinity for vinculin (initial design: 3.5 nM to 69 nM; mutant: 2.4 nM to >40µM affinity for vinculin) to reduce the dark state activity. </br></br> Lov-ipaA-VinD1 is a powerful tool which allows photocontroled complex formation. To establish this system Lungu et al. (2012)<sup><a href=#361>36.1</a></sup> fused the Ja helix of the LOV Domain to ipaA.</br>To be more precise they used the LOV2 domain from Avena sativa photopropin 1 (AsLOV2), which – as previously shown – can be used to photomodulate the affinity of peptides for their binding partners (see Figure 1). </br><div class=contant-image><img src=https://static.igem.org/mediawiki/2013/0/02/Bonn_MS_Figure1_LOV.jpg></br>Figure 1: General design of AsLOV2 fusion proteins (Lungu et al. 2012)<sup><a href=#361>36.1</a></sup></div> </br>In other studies had been shown that the LOV domain can be fused to entire protein domains, allowing photomodulation of the protein binding. However, they stated that it might be of high importance to bring the LOV domain closer to ipaA, in order to allow photomodulation in this case, because ipaA is only a peptide and thus more flexible than folded domains.</br></br>Therefore, Lungu et al. (2012)<sup><a href=361>36.1</a></sup> identified similar amino acid sequences in the ipaA peptide and the Ja helix of the LOV Domain and used this combined with molecular modeling to create photomodulateable AsLOV2-ipaA (see Figure 2). </br><div class=contant-image><img src=https://static.igem.org/mediawiki/2013/5/54/Bonn_MS_Figure2_LOV-ipaA.jpg></br>Figure 2: Light-inducible LOV-ipaA construct (Lungu et al. 2012)<sup><a href=361>36.1</a></sup></div></br>They were able to proof the functionality of the AsLOV2-ipaA system by heterodimerization in yeast (yeast two-hybrid system or Y2H) The yeast two-hybrid system can be used to monitor protein–protein interactions between two proteins. The system is based on a transcription factor, which is split into two separate fragments, called the binding domain (BD) and activating domain (AD). Each domain is fused to one protein and thus only if the proteins interact with each other BD and AD are close enough to initiate the transcription of a reporter gene.</br></br>The basic principle of the LOV-ipaA & VinD1 system works as follows. In the dark state the fusion product LOV-ipaA is not able to form a complex with vinculin, because LOV blocks ipaA sterically. However, activation of the LOV domain with blue light induces conformational changes in the fused molecule, which results in a movement of the Ja helix with the ipaA away from LOV. Thereby, ipaA becomes accessible for VinD1 and a Complex is formed.</br></br>The activation is reversible and the entire system can be genetically encoded. This two facts are the main advantages of this system in contrast to other typically used systems, which like for the chemical system for example, are based on in vivo covalently modified peptides, that can be activated by light induced cleavage. Moreover, the protein used are relatively small and thus should interfere as little as possible with the prokaryotic metabolism, the activity change form dark to light state is high, the system is completely genetically encoded and reversible. But, the most important property of this system is that it allows the light-controlled heterodimerisation of the two split variants of sspB, which is necessary for our system.</br></br><h2>References</h2></br><a name=361>36.1</a> <a href=http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3334866/>Lungu et al. (2012) Designing photoswitchable peptides using the AsLOV2 domain</a>";

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content.type="~~Project~~";

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content.type="Background";

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content.i = 37;

content.parents=[36,21,12];

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content.childs=[38,45,53,69,52,57,65,100];

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content.childs=[38,45,53,69,52,57,65,100,200];

content.titleShort = "Project summary";

content.titleLong = "Our project in a nutshell";

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content.summary= "We engineer a ~~photoswitchable~~ protein degradation system relying on the ClpXP protease system ~~and~~ a LOV domain from avena sativa";

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content.summary= "We engineer a photoswitachble protein degradation system relying on the ClpXP protease system an a LOV domain from avena sativa";

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content.text= "";

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content.text= "A reliable, yet easily adaptable mechanism for controlling protein activity is key to most areas of life and medical science research. Still, the most common approaches suffer from various flaws. Knocking genes out using homologous recombination, knocking a gene down with RNA interference or modulating the behaviour of a protein with a chemical stimulus - just to name a few prominent methods - is either restricted to non-lethal genes, does not yield a big difference in activity, or is absolutely inaccurate and thus prone to secondary effects.</br></br> Would it not be great if one could turn off any protein, at any time, with little to no side effects? That is where iGEM Bonn 2013 and their project comes in. We aim to overcome the aforementioned difficulties by engineering a novel tool based on blue-light-inducible degradation of targeted proteins.</br></br>Our system relies on two key components: A tiny (just 15 amino acids!) tag that is fused to the C-Terminus of a protein of your choosing, and a light sensing LOV (Light, Oxygen and Voltage) domain from Avena sativa.</br></br> The advantages of our approach are obvious: Not only does the usage of light allow for a superior tempero-spatial control, but it is also much less prone to unwanted side effects than any chemical stimulus.</br> Furthermore, as we rely on a direct degradation of the targeted protein, we expect an onset of the desired effect which is much faster and at least as high as in common approaches.</br> Finally, as our system requires only a minor modification of your target protein we expect its function to not be impaired, and the tag to go unnoticed in functional observations.";

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content.type="~~Project~~";

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content.type="Projekt";

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content.childs=[41,74,43];

content.titleShort = "C. crescentus";

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content.titleLong = "~~General information about~~ C. crescentus ~~and the ClpXP protein degradation system~~";

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content.titleLong = "Ortholog ClpXP system in C. crescentus";

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content.summary= "~~Here you can find a brief introduction to Caulobacter crescentus and its~~ ClpXP ~~protease~~ system.";

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content.summary= "Using the ortholog ClpXP protein degradation system of C. crescentus in E. coli enables us to induce protein degradation in native E. coli without interfering with their natural system.";

content.text= "Caulobacter crescentus is a Gram-negative α-protobacterium often found in fresh water lakes or in the sea. Its cell division cycle is a favoured object of study due to its remarkable asymmetry. <sup><a href=401>40.1</a></sup></br>ClpXP is a protease that degrades proteins tagged with the ssrA peptide. sspBα is an adaptor protein that can accelerate protein degradation by tethering ssrA-tagged proteins towards the ClpX subunit of the ClpXP protease.</br>For further information, please browse the related articles. </br></br><h2>References</h2></br><a href=401>40.1</a><a href=’ http://www.amazon.com/Brock-Biology-Microorganisms-13th-Edition/dp/032164963X’> Brock Microbiology of Microorganisms, Madigan et al., Pearson, German edition Vol. 13, 2012</a>";

content.type="Project";

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content.parents=[40];

content.childs=[];

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content.titleShort = "~~C. crescentus~~ ClpXP";

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content.titleShort = "<sup>Cc</sup>ClpXP";

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content.titleLong = "C. crescentus ClpXP";

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content.titleLong = "C. crescentus: ClpXP";

content.summary= "This article is about the ClpXP protease system in C. crescentus and regulation of proteolysis via ClpXP. It also provides information about similarities and differences between C. crescentus and E. coli orthologs of related proteins.";

content.text= "<div class='content-image'><img src='https://static.igem.org/mediawiki/2013/d/de/Bonn-41-CC-ClpXP.png'>ClpXP mediated proteolysis <sup><a href=#413>41.3</a></sup></div>ClpXP is an AAA+ protease which is of particular importance for proper cell-cycle progression. It consists of a hexameric ClpX subunit, which recognizes and unfolds tagged proteins, while ATP is hydrolyzed, and a ClpP subunit, that contains the actual peptidase domain. <sup><a href=#411>41.1</a>, <a href=#412>41.2</a></sup> Its structure is highly conserved, such that E. coli and C. crescentus orthologs are very similar. </br>Specific activity of the ClpXP protease is mediated by sspB and ssrA. ssrA is a short peptide consisting of fourteen amino acids in C. crescentus. Proteins which need to be degraded, e.g. for regulatory purpose or due to errors in their structure, are tagged with the ssrA peptide that can be recognized by the ClpX subunit. sspB is a dimeric adaptor protein in C. crescentus which tethers ssrA-tagged proteins to the ClpXP protease and, in this way, accelerates protein degradation. </br>Whilst sspB protein structure is well- conserved among many microorganisms, the structure of <sup>CC</sup>sspBα (i.e., the C. crescentus ortholog of sspB) shows significant differences compared to other orthologs. For example, <sup>CC</sup>sspBα and <sup>EC</sup>sspB (i.e., the E. coli ortholog) only show up with sequence identities of 16%, while CCsspB is still able to specifically bind to <sup>EC</sup>ClpXP<sup><a href=#412>41.2</a></sup>. SsrA tags can also be very different among microorganismic species. </br>This makes it possible to establish a protein degradation system in E. coli involving <sup>CC</sup>sspBα and <sup>EC</sup>ClpXP, as long as the ssrA tag can be recognized by both <sup>CC</sup>sspBα and <sup>EC</sup>ClpXP. </br></br><h2>References</h2></br><a name=411>41.1</a> <a href=’ http://www.ncbi.nlm.nih.gov/pubmed/17937918’>Structure and substrate specificity of an SspB ortholog: design implications for AAA+ adaptors, Chien et al., Cell Press, 2007, PMID: 17937918</a></br><a name=412>41.2</a> <a href=’http://www.ncbi.nlm.nih.gov/pubmed/20014030’>Versatile modes of peptide recognition by the ClpX N domain mediate alternative adaptor-binding specificities in different bacterial species, Chowdhury et al., Protein Science, 2010, PMID: 20014030</a></br><a name=413>41.3</a> <a href='http://www.biochem.umass.edu/faculty/peter-chien'>Peter Chien, Department of Biochemistry and Molecular Biology (official website), http://www.biochem.umass.edu/faculty/peter-chien, picture URL: cchttp://www.biochem.umass.edu/sites/biochem/files/resize/ClpX-300x268.jpg</a>";

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content.childs=[];

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content.titleShort = "~~SspB~~α";

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content.titleShort = "<sup>Cc</sup>sspBα";

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content.titleLong = "C. crescentus sspBα";

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content.titleLong = "C. crescentus: sspBα";

content.summary= "This article deals with the structure of sspBα and conformational details of its binding to ssrA and ClpXP during tethering.";

content.text= "<div class='content-image'><img src='https://static.igem.org/mediawiki/2013/thumb/4/42/Bonn_OutlookCCSspB1_Version2.png/726px-Bonn_OutlookCCSspB1_Version2.png'>sspB structure and its conservation among C. crescentus, E. coli and H. influenzae <sup><a href=#421>42.1</a></sup></div>The sspBα dimeric structure is stabilized by two α-helices in interaction, as part B of the figure above shows, each of them located at the N-terminus of either sspBα molecule. The subsequent parts of the protein form a domain consisting of two β-sheet structures, together building up the ssrA binding site. An unstructured area at the C-terminus being referred to as the XB module forms the ClpX binding part of the protein. It is connected to the rest of the molecule via a linker domain. <sup><a href=#422>42.2</a></sup> </br>Chien et al. <sup><a href=#421>42.1</a></sup> compared crystal structures of C. crescentus sspBα and its E. coli and H. influenzae sspB orthologs, discovering that in sspBα the α-helices are significantly longer, more twisted and cover a larger cross section area than the other two sspB orthologs. Also considering that β-sheets are rotated by around 20° in comparison to E. coli and H. influenzae orthologs, this leads to an antiparallel orientation of the two ssrA tagged protein bound to the ssrA binding sites of an sspBα dimer in C. crescentus, while they are parallel in γ-protobacterial sspB.</br></br><div class='content-image'><img src='https://static.igem.org/mediawiki/2013/6/6c/Bonn_OutlookCCSspB2.png' align=left>By measuring GFP fluorescence intensity, decrease of GFP-<sup>CC</sup>ssrA concentration (1) without sspBα added, (2) with mutated sspBα(Q74A) added , (3) with wildtype sspBα added can be visualized. <sup><a href=#421>42.1</a></sup></div>Chien et al. point out that although there are the remarkable differences in protein structure between sspBα and its γ-protobacterial ortholog, they show up with similar effectiveness in binding proteins tagged with the related ssrA peptide. But it turned out in their research that effectiveness of sspBα binding to the protein which needs to be tethered to the ClpXP protease strongly depends on which ssrA ortholog the protein is tagged with. sspBα binds firmly to <sup>CC</sup>ssrA, with an affinity being 175 times as large as for binding to <sup>EC</sup>ssrA (i.e. the E. coli ortholog). By comparing the crystal structures of both sspBα and the compound of sspBα and <sup>CC</sup>ssrA, Chien et al. further proved that binding of sspBα to <sup>CC</sup>ssrA does not lead to significant changes of its 3D conformation.</br></br><h2>References</h2></br><a name=421>42.1</a> <a href='http://www.ncbi.nlm.nih.gov/pubmed/17937918'>Structure and substrate specificity of an SspB ortholog: design implications for AAA+ adaptors, Chien et al., Cell Press, 2007, PMID: 17937918</a></br><a name=422>42.2</a><a href='http://www.ncbi.nlm.nih.gov/pubmed/14967151'> Bivalent tethering of sspB to ClpXP is required for efficient substrate delivery: a protein design study, Bolon DN et al., Mol Cell, 2004, PMID: 14967151</a>";

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content.parents=[40];

content.childs=[];

-

content.titleShort = "~~SspB~~ Split";

+

content.titleShort = "<sup>Cc</sup>sspB Split";

-

content.titleLong = "~~SspB~~ Split in C. crescentus";

+

content.titleLong = "sspB Split in C. crescentus";

content.summary= "The protein degradation system in Caulobacter crescentus resembles the system in E. coli, but the respective sequences of ssrA and SspB differ. Thus the different specifities can be used to introduce the ccSspB split system in wildtyp E. coli without disturbing the native processes in it.";

content.text= "The protein degradation system in Caulobacter crescentus resembles the system in E. coli, but the respective sequences of ssrA and SspB differ <sup><a href=#431>43.1</a></sup>. Thus ccssrA only binds ccSspB but not E. coli SspB. <sup><a href=#432>43.2</a></sup> <sup><a href=#433>43.3</a></sup> <sup><a href=#434>43.4</a></sup> However, proteins tagged with ccssrA can be degraded by E. coli ClpXP. Therefore the utilization of ccSspB and ccssrA in E. coli has the advantage that SspB+ strains can be used. <sup><a href=#431>43.1</a></sup> </br> In order to use this with the SspB split system, the fusion proteins ccSspBΔ10-FRB and FKBP12-SspB[XB] (E. coli) were incubated with GFP-ccDAS+4 and E. coli ClpXP in vitro. Without rapamycin there was no degradation detected. Equally, addition of E. coli SspB showed no degradation. Addition of rapamycin led to a reduction of GFP-ccDAS+4 of around 12% in 180 seconds. Compared to the E. coli split system (around 30 % in 180 seconds) this system is less fast but can, at least in vitro, be used with sspB-wildtype E. coli <sup><a href=#431>43.1</a></sup>. </br> As the results of the E.coli and the C. crescentus system in vitro show many similarities and the E. coli system works in vivo. It may be possible to use the C. crescentus in vivo as well. </br> <img src='https://static.igem.org/mediawiki/2013/8/82/Bonn-ccSspB.jpg'> <sup><a href=#431>43.1</a></sup> <h2>References:</h2> </br> <a name=431>43.1</a> <a href='http://dspace.mit.edu/bitstream/handle/1721.1/58089/654116495.pdf?sequence=1'> Understanding and Harnessing Energy-Dependent Proteolysis for Controlled Protein Degradation in Bacteria, J. Davis, Massachusetts Institute of Technology, april 2010 </a> </br> <a name=432>43.2</a> <a href='http://www.ncbi.nlm.nih.gov/pmc/articles/PMC58509/'> Overlapping recognition determinants within the ssrA degradation tag allow modulation of proteolysis, Flynn et al, Proc Natl Acad Sci USA 2001 Sep 11, PMID: 11535833 </a> </br> <a name=433>43.3</a> <a href='http://www.ncbi.nlm.nih.gov/pubmed/17937918'> Structure and substrate specifity of an SspB ortholog: design implications for AAA+ adaptors, Chien et al, Structure 2007 Oct, PMID: 17937918 <a/> </br> <a name=434>43.4</a> <a href='http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2581644/'> Inducible protein degradation in Bacillus subtilis using heterologous peptide tags and adaptor proteins to target substrates to the protease ClpXP, Griffith and Grossman, Mol Microbiol. 2008 Nov, PMID: 18811726 <a/> </br>";

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content.parents=[74];

content.childs=[];

-

content.titleShort = "SsrA";

+

content.titleShort = "<sup>Cc</sup>SsrA";

content.titleLong = "C. crescentus ssrA and its application in E. coli";

content.summary= "Proteins that need to be degraded by the ClpXP protease have to be tagged with the ssrA peptide previosly. Here is some information on the structure of ssrA.";

Line 501:

Line 503:

content.i =52;

content.parents=[37];

-

content.childs=[70];

+

content.childs=[];

content.titleShort="Achievements";

content.titleLong="Achievements";

Line 526:

Line 528:

content.childs=[67, 68];

content.titleShort = "Kill-switch for Lab Safety";

-

content.titleLong = "Kill-switch systems using ~~stress-induced~~ toxin-antitoxin modules ~~in Escherichia coli~~";

+

content.titleLong = "Light inducible Kill-switch systems using toxin-antitoxin modules";

content.summary= "Toxin-antitoxin systems are composed by an antitoxin and a toxin coding gene. Connecting our light inducible protein degradation system to the antitoxin via an ssrA-tag allows light induced cell death, as predominance of the toxin in a bacterium activates a cell death pathway.";

content.text= "Our system of light inducible protein degradation can be utilized to degrade any specific protein and is thus usable in a light induced kill-switch system. For this application a connection between the degradation system and a toxin-antitoxin module like MazEF or ccdA/ccdB is needed. Either the toxin or the antitoxin could be light inducibly degraded by adding an ssrA-tag, which is detected by our degradation system, to its genetical code: <ul><li><b>Using the degradation of the toxin:</b> For that purpose the insertion of a plasmid containing the ssrA-tagged toxin encoding gene is needed. Since the predominance of the toxin activates a cell death pathway in bacteria, a bacterium containing a module that allows light inducible degradation of the toxin would only be viable, when the toxin is degraded. In darkness the toxin overexpression is no longer compensated and aggregation of it leads to cell death. Apart from the use in lab security such a kill-switch system would also be useful for environmental applications of bacteria, since it opens up the possibility of deploying bacteria for only one day and ensures their passing by nightfall. For example bacteria could be used to perform the ecological stabilization of a lake but after one night any genetically modified bacteria would be dead.</li><li><b>Using the degradation of the antitoxin:</b> Two plasmids are needed: The first one to express the toxin and the second one to express the ssra-tagged antitoxin, in such manner that the amounts of the toxin and the antitoxin are in equilibrium. Once light induces the degradation system, the antitoxin is degraded and the predominant toxin will kill the bacterium. </li></ul> Regarding our idea to improve lab security by implementing a kill-switch system, both described ways seem possible. Usage of the former would require cultivating and working with the bacteria under constant blue light, as darkness would kill them. Realization of the latter would require no usage of any blue light in the lab since bacteria which get into touch with daylight our any blue light would be killed. Due to the high light sensivity of our degradation system it can most likely be induced by daylight, which renders the former killswitch system useless. Bacteria which escape from the lab could survive simply through contact with daylight. Consequently we focused on the second system (the degradation of the antitoxin).</br> With MazEF we described a light inducible kill-switch system via the insertion of plasmids into bacteria. However a final kill-switch system would have to be introduced into the genomic DNA since plasmids in bacteria can be ejected, for instance via cell division, whereas a genomic DNA mutation is less likely to occur. Nevertheless the risk of a loss of function cannot be eliminated , which is why a secure system should countain much more than one kill-switch system to compensate the malfunction of a single kill-switch system. Therefore, we consider the MazEF kill-switch system to be part of a much larger security system for genetically engineered bacteria. </br>";

Line 548:

Line 550:

content.i =57;

content.parents=[37];

-

content.childs=[~~58,59,60,61,62,63,64~~];

+

content.childs=[];

content.titleShort = "Results";

content.titleLong = "Results";

content.summary= "Results";

-

~~content.text="Results";~~

-

~~content.type="Project";~~

-

~~break;~~

-

~~case 58:~~

-

~~content.i =58;~~

-

~~content.parents=[57];~~

-

~~content.childs=[];~~

-

~~content.titleShort = "Expression mCherry";~~

-

~~content.titleLong = "Expression mCherry";~~

-

~~content.summary= "";~~

-

~~content.text="";~~

-

~~content.type="Project";~~

-

~~break;~~

-

~~case 59:~~

-

~~content.i =59;~~

-

~~content.parents=[57];~~

-

~~content.childs=[];~~

-

~~content.titleShort = "mCherry Abbau";~~

-

~~content.titleLong = "mCherry Abbau";~~

-

~~content.summary= "";~~

-

~~content.text="";~~

-

~~content.type="Project";~~

-

~~break;~~

-

~~case 60:~~

-

~~content.i =60;~~

-

~~content.parents=[57];~~

-

~~content.childs=[];~~

-

~~content.titleShort = "Rapamycin Abbauinduktion";~~

-

~~content.titleLong = "Rapamycin Abbauinduktion";~~

-

~~content.summary= "";~~

-

~~content.text="";~~

-

~~content.type="Project";~~

-

~~break;~~

-

~~case 61:~~

-

~~content.i =61;~~

-

~~content.parents=[57];~~

-

~~content.childs=[];~~

-

~~content.titleShort = "pDawn";~~

-

~~content.titleLong = "pDawn";~~

-

~~content.summary= "";~~

-

~~content.text="";~~

-

~~content.type="Project";~~

-

~~break;~~

-

~~case 62:~~

-

~~content.i =62;~~

-

~~content.parents=[57];~~

-

~~content.childs=[];~~

-

~~content.titleShort = "ccdB killswitch";~~

-

~~content.titleLong = "ccdB killswitch";~~

-

~~content.summary= "";~~

-

~~content.text="";~~

-

~~content.type="Project";~~

-

~~break;~~

-

~~case 63:~~

-

~~content.i =63;~~

-

~~content.parents=[57];~~

-

~~content.childs=[];~~

-

~~content.titleShort = "MazF killswitch";~~

-

~~content.titleLong = "MazF killswitch";~~

-

~~content.summary= "";~~

-

~~content.text="";~~

-

~~content.type="Project";~~

-

~~break;~~

-

~~case 64:~~

-

~~content.i =64;~~

-

~~content.parents=[57];~~

-

~~content.childs=[];~~

-

~~content.titleShort = "LOV degradation induction";~~

-

~~content.titleLong = "LOV degradation induction";~~

-

~~content.summary= "";~~

content.text="";

content.type="Project";

Line 637:

Line 562:

content.parents=[37];

content.childs=[];

-

content.titleShort = "~~Generated plasmids~~";

+

content.titleShort = "Plasmids";

-

content.titleLong = "~~Generated~~ plasmids";

+

content.titleLong = "A list of generated plasmids";

-

content.summary= "";

+

content.summary= "A list of each and every plasmid we designed combined with short summaries of their function";

-

~~content.text="";~~

+

content.text="<groupparts></groupparts></br></br><a href='https://static.igem.org/mediawiki/2013/e/ef/Bonn_AraC-pBAD%28D%29-RBS32-mazF-TT_BBa_K1075040.jpg'><img src='https://static.igem.org/mediawiki/2013/e/ef/Bonn_AraC-pBAD%28D%29-RBS32-mazF-TT_BBa_K1075040.jpg' width='700px'></a>MazF is part of the biological toxin-antitoxin system MazEF. This system is derived from the bacterium Escherichia coli, where it serves as a stress inducable killswitch. MazF, a RNA-degrading enzyme, ist he toxin and is normally inhibited by the presence of MazE. Absence of MazE leads to cell death.<sup><a href=http://parts.igem.org/Part:BBa_K1075040>1</a></sup><a href='https://static.igem.org/mediawiki/2013/3/34/Bonn_pLac1-RBS32-mazF-TT_BBa_K1075039.jpg'><img src='https://static.igem.org/mediawiki/2013/3/34/Bonn_pLac1-RBS32-mazF-TT_BBa_K1075039.jpg' width='700px'></a>We used this part to gain an IPTG and Lactose inducible construct of MazF. This part was used to test the toxicity of MazF as part of our proof of concept. <sup><a href=http://parts.igem.org/Part:BBa_K1075039>2</a></sup><a href='https://static.igem.org/mediawiki/2013/8/87/Bonn_pLac2-RBS32-mazE-%28ec%29ssrA%28DAS%2B4%29-TT_BBa_K1075043.jpg'><img src='https://static.igem.org/mediawiki/2013/8/87/Bonn_pLac2-RBS32-mazE-%28ec%29ssrA%28DAS%2B4%29-TT_BBa_K1075043.jpg' width='700px'></a>This part can be used to gain a Lactose and IPTG inducable construct of mazE-(ec)ssrA(DAS+4). We used this construct as part of our proof of concept. <sup><a href=http://parts.igem.org/Part:BBa_K1075043>3</a></sup><a href='https://static.igem.org/mediawiki/2013/c/cf/Bonn_AraC-pBAD-RBS-mazF-TT-pLac2-RBS-mazE-%28ec%29ssrA%28DAS%2B4%29-TT_BBa_K1075048.jpg'><img src='https://static.igem.org/mediawiki/2013/c/cf/Bonn_AraC-pBAD-RBS-mazF-TT-pLac2-RBS-mazE-%28ec%29ssrA%28DAS%2B4%29-TT_BBa_K1075048.jpg' width='700px'></a>This construct can be used to gain IPTG and Lactose inducible expression of mazE(ec)ssrA(DAS+4) and arabinose inducible expression of MazF. Thus, both parts of the toxin-antitoxin system mazEF are inducible and this construct can be used to further examine this system. We used this construct as part of our proof of concept.<sup><a href=http://parts.igem.org/Part:BBa_K1075048>4</a></sup><a href='https://static.igem.org/mediawiki/2013/7/7f/Bonn_AraC-pBAD-RBS-sspB-TT-pLac2-RBS-mCherry-TT_BBa_K1075049.jpg'><img src='https://static.igem.org/mediawiki/2013/7/7f/Bonn_AraC-pBAD-RBS-sspB-TT-pLac2-RBS-mCherry-TT_BBa_K1075049.jpg' width='700px'></a>The sspB protein is an adaptor responsible for delivering ssrA-tagged substrates to the ClpXP protease in order to enhance their degradation. mCherry is a red fluorescent protein with the excitation maximum at 587 nm and the Emission maximum at 610 nm.<sup><a href=http://parts.igem.org/Part:BBa_K1075049>5</a></sup><a href='https://static.igem.org/mediawiki/2013/4/42/Bonn_pDawn_%28Prom_%28const.%29-RBS34-YF1-FixJ-FixK-LambdaC-pC%29_BBa_K1075044.jpg'><img src='https://static.igem.org/mediawiki/2013/4/42/Bonn_pDawn_%28Prom_%28const.%29-RBS34-YF1-FixJ-FixK-LambdaC-pC%29_BBa_K1075044.jpg' width='700px'></a>The plasmid pDawn was designed by Ohlendorf et al. in 2012 together with its counter plasmid pDusk. Both plasmids are single plasmid systems, which allow the activation (pDawn) or repression (pDusk) of gene expression by blue light. They are easy to implement in the laboratory and lead to up to 460-fold activity change upon ilumination. <sup><a href=http://parts.igem.org/Part:BBa_K1075044>6</a></sup><a href='https://static.igem.org/mediawiki/2013/5/50/Bonn_pDawn_%28Prom_%28const.%29-RBS34-YF1-FixJ-FixK-LambdaC-pC%29-RBS32-ccdB-TT_BBa_K1075047.jpg'><img src='https://static.igem.org/mediawiki/2013/5/50/Bonn_pDawn_%28Prom_%28const.%29-RBS34-YF1-FixJ-FixK-LambdaC-pC%29-RBS32-ccdB-TT_BBa_K1075047.jpg' width='700px'></a>pDawn-RBS32-ccdB-TT can be used to photocontrol the expression of the toxin ccdB. In this system pDawn (BBa_K1075044) can be regulated by blue light, while ccdB (BBa_K1075029) acts as a killswitch. The ccd operon of the F plasmid encodes CcdB, a toxin targeting the essential gyrase of Escherichia coli, and CcdA, the unstable antidote that interacts with CcdB to neutralize its toxicity.<sup><a href=http://parts.igem.org/Part:BBa_K1075047>7</a></sup><a href='https://static.igem.org/mediawiki/2013/0/03/Bonn_AraC-pBAD%28D%29-RBS32-SspB%28Core%28-LOV-ipaA-TT_BBa_K1075019.jpg'><img src='https://static.igem.org/mediawiki/2013/0/03/Bonn_AraC-pBAD%28D%29-RBS32-SspB%28Core%28-LOV-ipaA-TT_BBa_K1075019.jpg' width='700px'></a>The part can be used to regulate the dimerization of the two parts of the split sspB. The binding of ipaA to Vincolin is regulated via the light-sensitive LOV domain. Therefore the two sspB parts only come together when the LOV domain was activated with blue light. <sup><a href=http://parts.igem.org/Part:BBa_K1075019>8</a></sup><a href='https://static.igem.org/mediawiki/2013/d/d0/Bonn_J23105-RBS32-SspB%28Core%29-LOV-ipaA-TT_BBa_K1075018.jpg'><img src='https://static.igem.org/mediawiki/2013/d/d0/Bonn_J23105-RBS32-SspB%28Core%29-LOV-ipaA-TT_BBa_K1075018.jpg' width='700px'></a>The part can be used to regulate the dimerization of the two parts of the split sspB. The binding of ipaA to Vincolin is regulated via the light-sensitive LOV domain. Therefore the two sspB parts only come together when the LOV domain was activated with blue light. <sup><a href=http://parts.igem.org/Part:BBa_K1075018>9</a></sup><a href='https://static.igem.org/mediawiki/2013/5/5a/Bonn_AraC-pBAD%28D%29-RBS32-VinD1-sspBXB-TT_BBa_K1075016.jpg'><img src='https://static.igem.org/mediawiki/2013/5/5a/Bonn_AraC-pBAD%28D%29-RBS32-VinD1-sspBXB-TT_BBa_K1075016.jpg' width='700px'></a>The part can be used to regulate the dimerization of the two parts of the split sspB. As Vincolin bins to ipaA, the sspB dimer is stabilized. We used this part to achieve a light-induced dimerization of the sspB parts. <sup><a href=http://parts.igem.org/Part:BBa_K1075016>10</a></sup><a href='https://static.igem.org/mediawiki/2013/9/9c/Bonn_J23118-RBS32-VinD1-SspBXB-TT_BBa_K1075015.jpg'><img src='https://static.igem.org/mediawiki/2013/9/9c/Bonn_J23118-RBS32-VinD1-SspBXB-TT_BBa_K1075015.jpg' width='700px'></a>The part can be used to regulate the dimerization of the two parts of the split sspB. As Vincolin bins to ipaA, the sspB dimer is stabilized. We used this part to achieve a light-induced dimerization of the sspB parts. <sup><a href=http://parts.igem.org/Part:BBa_K1075015>11</a></sup><a href='https://static.igem.org/mediawiki/2013/7/78/Bonn_pLac_pBAD2-RBS32-SspB-TT_BBa_K1075006.jpg'><img src='https://static.igem.org/mediawiki/2013/7/78/Bonn_pLac_pBAD2-RBS32-SspB-TT_BBa_K1075006.jpg' width='700px'></a>The sspB protein is an adaptor responsible for delivering ssrA-tagged substrates to the ClpXP protease in order to enhance their degradation. Prefixed is RBS, the binding site for a ribosome and the pLac promoter. It also includes a double terminator 'TT', which interrupts the translation. <sup><a href=http://parts.igem.org/Part:BBa_K1075006>12</a></sup><a href='https://static.igem.org/mediawiki/2013/8/8f/Bonn_E._coli_ssrA%28DAS%2B4%29-TT_BBa_K1075021.jpg'><img src='https://static.igem.org/mediawiki/2013/8/8f/Bonn_E._coli_ssrA%28DAS%2B4%29-TT_BBa_K1075021.jpg' width='700px'></a>The (ec)ssrA(DAS+4) tag can be fused to proteins so that they will be delivered to the ClpXP protease and degraded.<sup><a href=http://parts.igem.org/Part:BBa_K1075021>13</a></sup><a href='https://static.igem.org/mediawiki/2013/6/63/Bonn_AraC-pBad-RBS34-mCherry-ecssrA%28DAS%2B4%29-TT_BBa_K1075025.jpg'><img src='https://static.igem.org/mediawiki/2013/6/63/Bonn_AraC-pBad-RBS34-mCherry-ecssrA%28DAS%2B4%29-TT_BBa_K1075025.jpg' width='700px'></a>As we want to control protein degradation by controlling the function of ecSspB, we tagged the red fluorescent protein mCherry with ecssrA(DAS+4) to measure the degradation rate. The part was designed for proof of principle. Application as a bacterial fotographic film might be possible as well. <sup><a href=http://parts.igem.org/Part:BBa_K1075025>14</a></sup><a href='https://static.igem.org/mediawiki/2013/6/63/Bonn_AraC-pBad-RBS34-mCherry-ecssrA%28DAS%2B4%29-TT_BBa_K1075025.jpg'><img src='https://static.igem.org/mediawiki/2013/6/63/Bonn_AraC-pBad-RBS34-mCherry-ecssrA%28DAS%2B4%29-TT_BBa_K1075025.jpg' width='700px'></a>As we want to control protein degradation by controlling the function of ecSspB, we tagged the red fluorescent protein mCherry with ecssrA(DAS+4) to measure the degradation rate. The part was designed for proof of principle. Application as a bacterial fotographic film might be possible as well. <sup><a href=http://parts.igem.org/Part:BBa_K1075025>15</a></sup><a href='https://static.igem.org/mediawiki/2013/c/cd/Bonn_pJD427%28pC-RBS32-FKBP12-EcsspB%28XB%29-Term-pB-RBS32-EcsspB%28Core%29-FRB-Term-Term%29_BBa_K1075010.jpg'><img src='https://static.igem.org/mediawiki/2013/c/cd/Bonn_pJD427%28pC-RBS32-FKBP12-EcsspB%28XB%29-Term-pB-RBS32-EcsspB%28Core%29-FRB-Term-Term%29_BBa_K1075010.jpg' width='700px'></a>The SspB protein is an adaptor responsible for delivering ssrA-tagged substrates to the ClpXP protease in order to enhance their degradation. To control degradation it is reasonable to control the function of SspB. That is made by splitting it into two parts, each of which cannot induce degradation on its own. To bring both SspB parts together again for inducible degradation, they were combined with a chemical inducible heterodimerisation system: FRB and FKBP12. These two parts interact in the presence of rapamycin <sup><a href=http://parts.igem.org/Part:BBa_K1075010>16</a></sup><a href='https://static.igem.org/mediawiki/2013/d/d6/Bonn_%28pJD427%29-pLac-RBS34-mCherry-ecssrA%28DAS%2B4%29-TT_BBa_K1075027.jpg'><img src='https://static.igem.org/mediawiki/2013/d/d6/Bonn_%28pJD427%29-pLac-RBS34-mCherry-ecssrA%28DAS%2B4%29-TT_BBa_K1075027.jpg' width='700px'></a>As we want to control protein degradation by controlling the function of ecSspB, we tag the red fluorescent protein mCherry with ecssrA(DAS+4) to measure the degradation rate. We control ecSspB by splitting it into two parts each of which cannot induce degradation on its own but regains function by addition of rapamycin. As the co-transformation of the two plasmids didn’t work, we combined them to one.<sup><a href=http://parts.igem.org/Part:BBa_K1075027>17</a></sup><a href='https://static.igem.org/mediawiki/2013/b/bc/Bonn_pJD427-pLac-RBS34-mCherry-TT_BBa_K1075028.jpg'><img src='https://static.igem.org/mediawiki/2013/b/bc/Bonn_pJD427-pLac-RBS34-mCherry-TT_BBa_K1075028.jpg' width='700px'></a>This plasmid is a negative control to the pJD427-pLac2-RBS34-mCherry-ecssrA(DAS+4)-TT plasmid. <sup><a href=http://parts.igem.org/Part:BBa_K1075028>18</a></sup><a href='https://static.igem.org/mediawiki/2013/c/cd/Bonn_pJD427%28pC-RBS32-FKBP12-EcsspB%28XB%29-Term-pB-RBS32-EcsspB%28Core%29-FRB-Term-Term%29_BBa_K1075010.jpg'><img src='https://static.igem.org/mediawiki/2013/c/cd/Bonn_pJD427%28pC-RBS32-FKBP12-EcsspB%28XB%29-Term-pB-RBS32-EcsspB%28Core%29-FRB-Term-Term%29_BBa_K1075010.jpg' width='700px'></a>The Plasmid pJD427 contains the fusion proteins SspB[CORE]-FRB and FKBP12-SspB[XB]: SspB[CORE]-FRB with the weak constitutive promoter proB, FKBP12-SspB[XB] with the strong constitutive promoter proC and a medium-copy p15a origin of replication. <sup><a href=http://parts.igem.org/Part:BBa_K1075010>19</a></sup><a href='https://static.igem.org/mediawiki/2013/e/e1/Bonn_pLac-RBS32-ccdA-ssrA-TT_BBa_K1075035.jpg'><img src='https://static.igem.org/mediawiki/2013/e/e1/Bonn_pLac-RBS32-ccdA-ssrA-TT_BBa_K1075035.jpg' width='700px'></a>CcdA inhibits the toxin ccdB by binding to it and thus represses cell death. The part contains ccdA with a ssrA tag under the control of the Lac operon. <sup><a href=http://parts.igem.org/Part:BBa_K1075035>20</a></sup><a href='https://static.igem.org/mediawiki/2013/8/81/Bonn_AraC-pBAD%28D%29-RBS32-ccdB-TT_BBa_K1075031.jpg'><img src='https://static.igem.org/mediawiki/2013/8/81/Bonn_AraC-pBAD%28D%29-RBS32-ccdB-TT_BBa_K1075031.jpg' width='700px'></a>The part contains a chemically induced kill-switch. When Arabinose is added to the bacteria, the toxin ccdB is expressed and cell death is initiated. <sup><a href=http://parts.igem.org/Part:BBa_K1075031>20</a></sup>";

content.type="Project";

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content.childs=[];

content.titleShort = "MazEF";

-

content.titleLong = "~~A kill-switch system using the stress-induced~~ toxin-antitoxin ~~module MazEF in Escherichia coli~~";

+

content.titleLong = "The maz toxin-antitoxin system";

content.summary= "The toxin-antitoxin system MazEF is composed by an upstream gene <i>mazE</i>, encoding a labile antitoxin, and a downstream gene <i>mazF</i>, that encodes a stable toxin. Connecting our light inducible protein degradation system to the antitoxin MazE allows light inducible cell death, as predominance of MazF in a bacterium activates cell death pathway.";

-

content.text= "Our system of light inducible protein degradation can be utilized to degrade any specific protein and is therefore usable to realize a light inducible kill-switch system. A connection of protein degradation to a cell death pathway is represented by the stress-induced toxin-antitoxin module <i>mazEF</i> in Escherichia coli. <i>mazEF</i> is located on the chromosome in E.coli which is associated with programmed cell death. The toxin-antitoxin system is composed by an upstream gene <i>mazE</i>, encoding a labile antitoxin, and a downstream gene mazF, that encodes a stable toxin. </br>The product of <i>mazF</i> cleaves mRNAs and tmRNAs at a specific site, which leads to an inhibition of translation. MazF shows a specific cleaving mechanism, which is not well understood yet, but shows that there is also protein synthesis which is unaffected by MazF. These proteins are presumably part of a cell death pathway. </br>The effect of <i>mazF</i> is suppressed by the Product of <i>mazE</i> which is degraded by the Protease ClpAP in bacteria. As a result of stressful conditions expression of the chromosomal <i>mazEF</i> module is reduced which leads to an imbalance between the products of <i>mazF</i> and <i>mazE</i>: When expression is lowered the stable toxin of <i>mazF</i> still persists while the labile antitoxin of <i>mazE</i> is degraded and can no longer suppress the effect of <i>mazF</i> leading to acute toxicity and cell death. </br><i>MazEF</i>-mediated cell death in E. coli can be caused by:<ul><li>extreme amino acid starvation<sup><a href = #671>67.1</a></sup></li><li> inhibition of transcription and/or translation by antibiotics such as rifampin, chloramphenicol, and spectinomycin under specific growth conditions<sup><a href = #671>67.1</a></sup></li><li>inhibition of translation by the Doc protein of prophage P1<sup><a href = #671>67.1</a></sup></li><li>DNA damage caused by thymine starvation as well as by mitomycin C, nalidixic acid, and UV irradiation<sup><a href = #671>67.1</a></sup></li><li>oxidative stress (H2O2)<sup><a href = #671'>67.1</a></sup></li></ul>Amitai et al. tested in 2004 the Hypothesis of Pedersen et al.<sup><a href = #672>67.2</a></sup>, that chromosomal toxin-antitoxin systems may rather cause a state of reversible bacteriostasis than programmed cell death<sup><a href = #671>67.1</a></sup>.Therefore E.coli strain MC4100 &#916<i>mazE</i>F relA1 lacIq was cotransformated with:<ul><li>pBad-<i>mazF</i></li><li>pQE-&#916his-<i>mazE</i></li></ul><i>mazF</i>-expression can be induced by the addition of Arabinose via the pBad promoter of the first plasmid. The transformation of the second plasmid results firstly in the repression of <i>mazE</i> expression, whereas when IPTG is added <i>mazE</i> production is induced.</br><div class='content-image' align='center' height=501 width=410><a href='https://static.igem.org/mediawiki/2013/a/ac/Team_Bonn_MazF_1.png'><img src='https://static.igem.org/mediawiki/2013/a/ac/Team_Bonn_MazF_1.png' height=491 width=400></a></br><i>Ability of E. coli cells that had been ectopically overexpressing MazF in liquid medium to form colonies when ectopically overexpressing MazE on plates. The cultures were grown in LB medium (A) or M9 minimal medium with 0.5% glycerol (B) at 37°C to midlogarithmic phase (OD600, 0.5)<sup><a href = #671>67.1</a></sup>.</i></div>Using these tools, Amitai et al. tested the effect of MazE overproduction on MazF-overproducing bacteria during growth in liquid medium.</br>The E.coli strain was incubated in LB medium. After <i>mazF</i> expression was induced by adding arabinose two samples were taken at several time points. To repress <i>mazF</i> expression to both of them glucose was added. In addition IPTG was added to one culture to induce <i>mazE</i> expression. The two cultures were compared via the level of protein synthesis and OD600.</br>Finally Amitai et al. confirmed the assumption that the overproduction of MazE after until 6h under overproduction of MazF could resuscitate E.coli cells in LB medium (Fig. 1A)<sup><a href = #671>67.1</a></sup>, but the longer MazF was induced the less cells could be resuscitated by MazE.</br>Whereas MazE overproduction can reverse the inhibitory effect of MazF on translation, it cannot reverse the effect of MazF on colony formation, which is shown in figure 2. Only 1h after the induction of MazE expression, the rate of translation was restored to nearly 100% (Fig.2 Aa, Ab, Ac) but the bacteriocidic effect could not be reversed (Fig.2 Ba, Bb, Bc).</br>Additionally, Amtai et al. found out, that in M9 medium MazE was less able to reverse the effects of MazF overexpression than in LB medium (Fig.1B vs. 1A). it was concluded that there is a point of no return, when MazE is inable to resuscitate a MazF damaged cell, which occurs earlier in M9 medium than in LB medium.</br>Based on their results a model of the MazEF mechanism was built: A <i>mazF</i>-mediated cascade leads to a cell death pathway, but can nevertheless be stopped at several intermediary steps by e.g. <i>mazE</i>. When a point of no return is reached, the cascade cannot be stopped anymore.<div class='content-image' align='center' height=827 width=784><a href='https://static.igem.org/mediawiki/2013/9/9e/Team_Bonn_MazF_2.png'><img src='https://static.igem.org/mediawiki/2013/9/9e/Team_Bonn_MazF_2.png' height=817 width=764></a></br><i>Effect of MazE overproduction during growth in liquid medium on the ability of MazF-overproducing E. coli cells to synthesize proteins and form colonies. To induce <i>mazE</i> expression, IPTG was added to the bacterial culture at 1 h (Aa and Ba), 4 h (Ab and Bb), and 6 h (Ac and Bc) after <i>mazF</i> induction at time zero. The effects of the ectopic overexpression of MazE were measured at 1 and 3 h after the induction of <i>mazE</i> expression.<sup><a href = #671>67.1</a></sup>.</i></div></br>Back to our project and to the idea of a light inducible kill-switch system:</br>As we described in the previous paragraph for both of our kill-switch systems using the MazEF module either MazF or MazE could be degraded:<ul><li>Using the degradation of MazF:</b> For that purpose the insertion of a plasmid containing the ssrA-tagged toxin encoding gene is needed. Since the predominance of the toxin activates a cell death pathway in bacteria, a bacterium containing a module that allows light inducible degradation of the MazF would only be viable, when it is degraded. In darkness the MazF overexpression is no longer compensated and aggregation of it leads to cell death. </li><li><b>Using the degradation of the antitoxin:</b> Two plasmids are needed: The first one to express the MazF and the second one to express the ssra-tagged MazE, in such manner that the amounts of the <i>MazF</i> and the <i>MazE</i> are in equilibrium. Once light induces the degradation system, the MazE is degraded and the predominant toxin will kill the bacterium.</li></ul>As we explained in the general kill-switch system text we finally focused on the second system (via the degradation of MazE).</br>With the design of a MazEF kill-switch system the possibility of resuscitating bacteria in the way Amitai et al. showed has to be considered. A predominant MazF could kill a bacterium in LB within about two hours, but it needs to be predominant over a long period (>7h) to induce its death without it being resuscitated by renewed <i>mazE</i> expression (Fig.1A)with more than 50% probability .</br>Certainly these facts seem to be unfavourable for the realization of a kill switch system via MazEF, but fortunately our system of heterodimerization (Lungu et al.) allows long continuous degradation<sup><a href = #673>67.3</a></sup>, due to the high stability of the light induced hererodimer. Therefore likely a short exposure time will result in prolonged protein degradation sufficing for bacterial death. Additionally, Amitai et al. showed that the less nutrition is available for a bacterium, the earlier the point of no return is reached. If a bacterium escapes the lab, it will likely have less nutrition available than in LB medium. It might reach the point of no return earlier.</br>We described a light inducible MazEF kill-switch system via the insertion of plasmids into bacteria. However a final kill-switch system would have to be implemented in the genomic DNA since plasmids in bacteria can be ejected, for instance via cell division, whereas a genomic DNA mutation is less likely to occur. Nevertheless risk of a loss of function cannot be eliminated , which is why a secure system should countain much more than one kill-switch system to compensate the malfunction of a single kill-switch system. Therefore, we consider the MazEF kill-switch system to be part of a much larger security system for genetically engineered bacteria.<h2>References</h2><a name =671>67.1</a> <a href = 'http://www.ncbi.nlm.nih.gov/pmc/articles/PMC532418/'>MazF-Mediated Cell Death in Escherichia coli: a Point of No Return, Shahar Amitai et al., Journal of Bacteriology Vol. 186, No. 24, 2004, p.8295–8300.</a></br><a name =672>67.2</a> <a href = 'http://www.ncbi.nlm.nih.gov/pubmed/?term=Rapid+induction+and+reversal+of+bacteriostatic+conditions+by+controlled+expression+of+toxins+and+antitoxins'>Rapid induction and reversal of bacteriostatic conditions by controlled expression of toxins and antitoxins, Pedersen et al., Molecular Microbiology 45, 2002, 501–510.</a></br><a name =673>67.3</a> <a href = 'http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3334866/'>Designing Photoswitchable Peptides Using the AsLOV2 Domain, Oana I. Lungu et al., Chem Biol. 2012, 19(4):507-17.</a>";

+

content.text= "Our system of light inducible protein degradation can be utilized to degrade any specific protein and is therefore usable to realize a light inducible kill-switch system. A connection of protein degradation to a cell death pathway is represented by the stress-induced toxin-antitoxin module <i>mazEF</i> in Escherichia coli. <i>mazEF</i> is located on the chromosome in E.coli which is associated with programmed cell death. The toxin-antitoxin system is composed by an upstream gene <i>mazE</i>, encoding a labile antitoxin, and a downstream gene mazF, that encodes a stable toxin. </br>The product of <i>mazF</i> cleaves mRNAs and tmRNAs at a specific site, which leads to an inhibition of translation. MazF shows a specific cleaving mechanism, which is not well understood yet, but shows that there is also protein synthesis which is unaffected by MazF. These proteins are presumably part of a cell death pathway. </br>The effect of <i>mazF</i> is suppressed by the Product of <i>mazE</i> which is degraded by the Protease ClpAP in bacteria. As a result of stressful conditions expression of the chromosomal <i>mazEF</i> module is reduced which leads to an imbalance between the products of <i>mazF</i> and <i>mazE</i>: When expression is lowered the stable toxin of <i>mazF</i> still persists while the labile antitoxin of <i>mazE</i> is degraded and can no longer suppress the effect of <i>mazF</i> leading to acute toxicity and cell death. </br><i>MazEF</i>-mediated cell death in E. coli can be caused by:<ul><li>extreme amino acid starvation<sup><a href = #674>67.4</a></sup><sup><a href = #675>67.5</a></sup></li><li> inhibition of transcription and/or translation by antibiotics such as rifampin, chloramphenicol, and spectinomycin under specific growth conditions<sup><a href = #676>67.6</a></sup></li><li>inhibition of translation by the Doc protein of prophage P1<sup><a href = #677>67.7</a></sup></li><li>DNA damage caused by thymine starvation<sup><a href = #678>67.8</a></sup> as well as by mitomycin C, nalidixic acid, and UV irradiation<sup><a href = #679>67.9</a></sup></li><li>oxidative stress (H2O2)<sup><a href = #679'>67.9</a></sup></li></ul>Amitai et al. tested in 2004 the Hypothesis of Pedersen et al.<sup><a href = #672>67.2</a></sup>, that chromosomal toxin-antitoxin systems may rather cause a state of reversible bacteriostasis than programmed cell death<sup><a href = #671>67.1</a></sup>.Therefore E.coli strain MC4100 &#916<i>mazE</i>F relA1 lacIq was cotransformated with:<ul><li>pBad-<i>mazF</i></li><li>pQE-&#916his-<i>mazE</i></li></ul><i>mazF</i>-expression can be induced by the addition of Arabinose via the pBad promoter of the first plasmid. The transformation of the second plasmid results firstly in the repression of <i>mazE</i> expression, whereas when IPTG is added <i>mazE</i> production is induced.</br><div class='content-image' align='center' height=501 width=410><a href='https://static.igem.org/mediawiki/2013/a/ac/Team_Bonn_MazF_1.png'><img src='https://static.igem.org/mediawiki/2013/a/ac/Team_Bonn_MazF_1.png' height=491 width=400></a></br><i>Ability of E. coli cells that had been ectopically overexpressing MazF in liquid medium to form colonies when ectopically overexpressing MazE on plates. The cultures were grown in LB medium (A) or M9 minimal medium with 0.5% glycerol (B) at 37°C to midlogarithmic phase (OD600, 0.5)<sup><a href = #671>67.1</a></sup>.</i></div>Using these tools, Amitai et al. tested the effect of MazE overproduction on MazF-overproducing bacteria during growth in liquid medium.</br>The E.coli strain was incubated in LB medium. After <i>mazF</i> expression was induced by adding arabinose two samples were taken at several time points. To repress <i>mazF</i> expression to both of them glucose was added. In addition IPTG was added to one culture to induce <i>mazE</i> expression. The two cultures were compared via the level of protein synthesis and OD600.</br>Finally Amitai et al. confirmed the assumption that the overproduction of MazE after until 6h under overproduction of MazF could resuscitate E.coli cells in LB medium (Fig. 1A)<sup><a href = #671>67.1</a></sup>, but the longer MazF was induced the less cells could be resuscitated by MazE.</br>Whereas MazE overproduction can reverse the inhibitory effect of MazF on translation, it cannot reverse the effect of MazF on colony formation, which is shown in figure 2. Only 1h after the induction of MazE expression, the rate of translation was restored to nearly 100% (Fig.2 Aa, Ab, Ac) but the bacteriocidic effect could not be reversed (Fig.2 Ba, Bb, Bc).</br>Additionally, Amtai et al. found out, that in M9 medium MazE was less able to reverse the effects of MazF overexpression than in LB medium (Fig.1B vs. 1A). it was concluded that there is a point of no return, when MazE is inable to resuscitate a MazF damaged cell, which occurs earlier in M9 medium than in LB medium.</br>Based on their results a model of the MazEF mechanism was built: A <i>mazF</i>-mediated cascade leads to a cell death pathway, but can nevertheless be stopped at several intermediary steps by e.g. <i>mazE</i>. When a point of no return is reached, the cascade cannot be stopped anymore.<div class='content-image' align='center' height=827 width=784><a href='https://static.igem.org/mediawiki/2013/9/9e/Team_Bonn_MazF_2.png'><img src='https://static.igem.org/mediawiki/2013/9/9e/Team_Bonn_MazF_2.png' height=817 width=764></a></br><i>Effect of MazE overproduction during growth in liquid medium on the ability of MazF-overproducing E. coli cells to synthesize proteins and form colonies. To induce <i>mazE</i> expression, IPTG was added to the bacterial culture at 1 h (Aa and Ba), 4 h (Ab and Bb), and 6 h (Ac and Bc) after <i>mazF</i> induction at time zero. The effects of the ectopic overexpression of MazE were measured at 1 and 3 h after the induction of <i>mazE</i> expression.<sup><a href = #671>67.1</a></sup>.</i></div></br>Back to our project and to the idea of a light inducible kill-switch system:</br>As we described in the previous paragraph for both of our kill-switch systems using the MazEF module either MazF or MazE could be degraded:<ul><li>Using the degradation of MazF:</b> For that purpose the insertion of a plasmid containing the ssrA-tagged toxin encoding gene is needed. Since the predominance of the toxin activates a cell death pathway in bacteria, a bacterium containing a module that allows light inducible degradation of the MazF would only be viable, when it is degraded. In darkness the MazF overexpression is no longer compensated and aggregation of it leads to cell death. </li><li><b>Using the degradation of the antitoxin:</b> Two plasmids are needed: The first one to express the MazF and the second one to express the ssra-tagged MazE, in such manner that the amounts of the <i>MazF</i> and the <i>MazE</i> are in equilibrium. Once light induces the degradation system, the MazE is degraded and the predominant toxin will kill the bacterium.</li></ul>As we explained in the general kill-switch system text we finally focused on the second system (via the degradation of MazE).</br>With the design of a MazEF kill-switch system the possibility of resuscitating bacteria in the way Amitai et al. showed has to be considered. A predominant MazF could kill a bacterium in LB within about two hours, but it needs to be predominant over a long period (>7h) to induce its death without it being resuscitated by renewed <i>mazE</i> expression (Fig.1A)with more than 50% probability .</br>Certainly these facts seem to be unfavourable for the realization of a kill switch system via MazEF, but fortunately our system of heterodimerization (Lungu et al.) allows long continuous degradation<sup><a href = #673>67.3</a></sup>, due to the high stability of the light induced hererodimer. Therefore likely a short exposure time will result in prolonged protein degradation sufficing for bacterial death. Additionally, Amitai et al. showed that the less nutrition is available for a bacterium, the earlier the point of no return is reached. If a bacterium escapes the lab, it will likely have less nutrition available than in LB medium. It might reach the point of no return earlier.</br>We described a light inducible MazEF kill-switch system via the insertion of plasmids into bacteria. However a final kill-switch system would have to be implemented in the genomic DNA since plasmids in bacteria can be ejected, for instance via cell division, whereas a genomic DNA mutation is less likely to occur. Nevertheless risk of a loss of function cannot be eliminated , which is why a secure system should countain much more than one kill-switch system to compensate the malfunction of a single kill-switch system. Therefore, we consider the MazEF kill-switch system to be part of a much larger security system for genetically engineered bacteria.<h2>References</h2><a name =671>67.1</a> <a href = 'http://www.ncbi.nlm.nih.gov/pmc/articles/PMC532418/'>MazF-Mediated Cell Death in Escherichia coli: a Point of No Return, Shahar Amitai et al., Journal of Bacteriology Vol. 186, No. 24, 2004, p.8295–8300.</a></br><a name =672>67.2</a> <a href = 'http://www.ncbi.nlm.nih.gov/pubmed/?term=Rapid+induction+and+reversal+of+bacteriostatic+conditions+by+controlled+expression+of+toxins+and+antitoxins'>Rapid induction and reversal of bacteriostatic conditions by controlled expression of toxins and antitoxins, Pedersen et al., Molecular Microbiology 45, 2002, 501–510.</a></br><a name =673>67.3</a> <a href = 'http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3334866/'>Designing Photoswitchable Peptides Using the AsLOV2 Domain, Oana I. Lungu et al., Chem Biol. 2012, 19(4):507-17.</a></br><a name =674>67.4</a> <a href = 'http://www.ncbi.nlm.nih.gov/pmc/articles/PMC39188/'>An Escherichia coli chromosomal addiction module regulated by guanosine 3,5-bispyrophosphate: a model for programmed bacterial cell death, Aizenman, E., H. Engelberg-Kulka, and G. Glaser, Proc. Natl. Acad. Sci., 1996, USA 93:6059-6063.</a></br><a name =675>67.5</a> <a href = 'http://www.ncbi.nlm.nih.gov/pmc/articles/PMC28068/'>rexB of bacteriophage lambda is an anti-cell death gene. Engelberg-Kulka, H., M. Reches, S. Narasimhan, R. Schoulaker-Schwarz, Y. Klemes, E. Aizenman, and G. Glaser, Proc. Natl. Acad. Sci., 1998, USA 95:15481-15486.</a></br><a name =676>67.6</a> <a href = 'http://www.ncbi.nlm.nih.gov/pmc/articles/PMC95100/'>Programmed cell death in Escherichia coli: some antibiotics can trigger mazEF lethality, Sat, B., R. Hazan, T. Fisher, H. Khaner, G. Glaser, and H. Engelberg-Kulka, J. Bacteriol. 2001, 183:2041-2045.</a></br><a name =677>67.7</a> <a href = 'http://www.ncbi.nlm.nih.gov/pmc/articles/PMC95101/'>Postsegregational killing mediated by the P1 phage addiction module Phd-Doc requires the Escherichia coli programmed cell death system mazEF, Hazan, R., B. Sat, M. Reches, and H. Engelberg-Kulka, J. Bacteriol. 2001, 183:2046–2050.</a></br><a name =678>67.8</a> <a href = 'http://www.ncbi.nlm.nih.gov/pmc/articles/PMC150121/'>The Escherichia coli mazEF suicide module mediates thymineless death, Sat, B., M. Reches, and H. Engelberg-Kulka, J. Bacteriol. 2003, 185:1803–1807.</a></br><a name =679>67.9</a> <a href = 'http://www.ncbi.nlm.nih.gov/pmc/articles/PMC415763/'>Escherichia coli mazEFmediated cell death is triggered by various stressful conditions, Hazan, R., B. Sat, and H. Engelberg-Kulka, J. Bacteriol. 2004, 186:3663–3669.</a>";

content.type="Project";

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content.parents=[54];

content.childs=[];

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content.titleShort = "~~ccdA/ccdB~~";

+

content.titleShort = "ccdAB";

content.titleLong = "The ccd toxin-antitoxin system";

-

content.summary= "~~this~~ is a ~~summary6~~";

+

content.summary= "CcdB leads to double strand breaks in bacterial genomic DNA and thus is a potent toxin leading to cell death. ccdA compromises the toxic effect of ccdB.";

content.text= "The ccd module is a toxin-antitoxin (TA) system similar to the mazE/mazF system. The module is located on the F Plasmid in Escherichia coli bacteria and essential for their survival. Normally the toxin ccdB is inactivated by the presence of the antitoxin ccdA in the form of a ccdAB complex. If ccdA is no longer available, ccdB inhibits DNA gyrase which leads to cell death. Gyrase is a type IIA topoisomerase and is able to produce negative DNA supercoiling by making a double-strand break in the DNA and religating it. The gyrase enzyme consists of two subunits: the C-terminal GyrA domain that wraps around the DNA strand and the N-terminal GyrB domain that catalyses the ATP-dependant supercoiling of the DNA. CcdB stabilizes the gyrase cleavage complex by binding to the GyrA domain and thus blocks the catalytic function of the gyrase. That means that the gyrase remains bound to the DNA and the cleaved DNA is not religated. DNA- and RNA polymerases can’t copy the DNA anymore and cell proliferation as well as protein biosynthesis is stopped. The double-stranded breaks in the DNA initiate cell death.<p>Because gyrases are specific to bacteria such as E. coli it is also a target for some anti-bacterial medications e.g. ciprofloxacin (CFX). As can be seen in the data below, CcdB proves to be as effective as CFX at inducing DNA cleavage <sup><a href=#681>68.1</a></sup><sup> <a href=#682>68.2</a></sup></p><div class='content-image'align='center'><img src='https://static.igem.org/mediawiki/2013/2/2f/Sspb_CFX_compared.jpg' width='300'>Comparison of the effect of CcdB and CFX on gyrase activity. N: negatively supercoiled DNA, L: linear DNA, SC: supercoiled DNA. A higher concentration of CcdB/CFX leads to more cleaved (linear) DNA<sup><a href= http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1635281/figure/fig2/>[source]</a></sup></div><div class='content-image' align='center'><img src='https://static.igem.org/mediawiki/2013/e/e4/Bonn_Ccdb_and_ccda.jpg' width='300'> A higher concentration of ccdB leads to blocking of gyrase and positively supercoiled DNA is cleaved to linear DNA instead of being processed to negatively supercoiled DNA. <sup><a href=http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3460896/figure/pone-0046499-g003/>[source]</a></sup></div><p>We used this system to build a light-induced kill-switch. Therefore we added the ssrA tag to the antitoxin ccdA. When the bacteria a emitted to light, ccdA is degraded and ccdB is set free and can bind to the gyrase. and cell death is initiated. Like in most TA systems, the toxin ccdB is relatively stable, while the antitoxin ccdA is vulnerable to degradation.</p><h2>References</h2><p><a name=681>68.1</a> <a href='http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1635281/'>A strand-passage conformation of DNA gyrase is required to allow the bacterial toxin, CcdB, to access its binding site, Andrew B. Smith and Anthony Maxwell, Nucleic Acids Res. 2006 October; 34(17): 4667–4676, PMC 1635281</a></p><p><a name=682>68.2</a> <a href='http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3460896/'> A Common Origin for the Bacterial Toxin-Antitoxin Systems parD and ccd, Suggested by Analyses of Toxin/Target and Toxin/Antitoxin Interactions, Andew B. Smith et al, PLoS One. 2012; 7(9): e46499, PMCID: PMC3460896</a></p>";

content.type="Project";

Line 691:

Line 617:

break;

-

~~case 70:~~

-

~~content.i =70;~~

-

~~content.parents=[52];~~

-

~~content.childs=[];~~

-

~~content.titleShort = "Timeline";~~

-

~~content.titleLong = "Timeline";~~

-

~~content.summary= "";~~

-

~~content.text="";~~

-

~~content.type="Project";~~

-

~~break;~~

case 71:

Line 753:

Line 669:

content.i = 100;

content.parents=[37];

-

content.childs=[106,107,108,~~109~~,101,110,130~~,112~~];

+

content.childs=[106,107,108,112,101,110,130];

content.titleShort = "Human Practice";

content.titleLong = "Human Practice in General";

-

content.summary= "~~The major goal in~~ Human Practice ~~is to make synthetic biology easily understandable and interesting for everybody. For that purpose~~ our project's design follows the well-known franchise of Star Wars; our project name "LOV WARS" was born. The constant references to the Star Wars movies facilitate the access to synthetic biology to a person that is not familiar with the subject~~. With the most famous icon of Star Wars being the laser sword it offers a great opportunity to present our light induction system in an entertaining way~~. We put great effort into adapting every aspect of our public presence to Star Wars.";

+

content.summary= "For our Human Practice advances our project's design follows the well-known franchise of Star Wars - being inspired by the LOV domain our project name "LOV WARS" was born. The constant references to the Star Wars movies facilitate the access to synthetic biology to a person that is not familiar with the subject. We put great effort into adapting every aspect of our public presence to Star Wars.";

content.text= "<p>The major goal in Human Practice is to make synthetic biology easily understandable and interesting for everybody. For that purpose our project's design follows the well-known franchise of Star Wars; our project name 'LOV WARS' was born. The constant references to the Star Wars movies facilitate the access to synthetic biology to a person that is not familiar with the subject. With the most famous icon of Star Wars being the laser sword it offers a great opportunity to present our light induction system in an entertaining way. We put great effort into adapting every aspect of our public presence to Star Wars.</p><p>Therefore we introduced a <a onclick=node(110)>flash game</a> and a <a onclick=node(109)>comic series</a> called LOV Wars in which we explain basic concepts of synthetic biology and our project. The comic and the LOV Wars shooter arouse interest in people that would normally never engage with the field. That way we can spread the possibilities and advantages that modern genetically engineered organisms offer and clear possible misunderstandings or prejudices against the subject.</p>For the same purpose we organized multiple events for general public at which we presented the tools used in synthetic biology and our project; over the last six months we <a onclick=node(108)>visited several High Schools</a> in Bonn and nearby and gave presentations. In addition we had an information booth in our city center informing passersby about the risks and benefits of synthetic biology with the aid of several posters and flyers. A big highlight was a <a onclick=node(106)>Science slam</a> we arranged. In front of a big audience six scientists from different fields presented an interesting aspect of their academic field. The Science Slam was a great success in terms of getting people in touch with science and without exception received very good critiques.<br> At every opportunity we handed out questionnaires in which we evaluated the public's opinion about synthetic biology and whether our human practice events were informative and interesting.";

content.type="Human Practice";

Line 765:

Line 681:

content.parents=[100];

content.childs=[102,103];

-

content.titleShort = "~~Project presentations~~"

+

content.titleShort = "Presentations"

content.titleLong = "Project presentations";

content.summary= "We presented iGEM and our project on two scientific conferences.";

Line 787:

Line 703:

content.parents=[101];

content.childs=[];

-

content.titleShort = "~~Biocom congress~~ Berlin";

+

content.titleShort = "Congress Berlin";

content.titleLong = "Biocom congress Berlin";

content.summary= "our day in Berlin. We met important people and other iGEM teams";

Line 812:

Line 728:

content.titleLong = "The Science Slam - Science for Everybody";

content.summary = "We organized a science slam to offer an experience of science for everyone. And the event met with great approval!";

-

content.text = "<h1>Science Slam</h1><p>Our project aims to make versatile control of biological machines easily possible for everybody. During conception we wondered how our human practice work could represent that thought and decided to organize a science slam an event which makes science available to everybody!</p><p>We believe that science must not only stay in laboratories and lecture halls but reach lots of people out there. And without a doubt, the walls of the reputed ivory tower couldnt stand throughout the evening of our event!</p><p>Furthermore, the science slam provided a platform for our slammers to share the passion for their project with more than just their colleagues and thus help to popularize their field. In ten minutes, they presented their topic in a playful and inspirational manner and the audience decided which slammer has managed to get them carried away the most.</p><p>We managed not only to give floor to a large variety of fields by our slammers from physics, medicine, psychology, informatics, politics and engineering but to make them be heard by a public which was very mixed indeed.</p><p>As a prelude to the event, we acquainted the audience with synthetic biology and our project by showing what synthetic biology can do for each of us. Distributed questionnaires during the evening showed that -as hoped- our presentation helped to leave a positive mark on the field of synthetic biology in ~~peopleŽs~~ heads:</p><p>For example, 97% of the people, who stated they did not know about synthetic biology before have got a proper picture of it during our event. 86% said, they are now familiar with the iGEM competition and everybody stated, they found the idea to combine the presentation of our iGEM project with a science slam very good or ~~good~~.</p><p>The fact that our audience liked this topic had been reflected in our voting: Our representative for synthetic biology, Mogan Ramesh won the science slam. He originally comes from engineering but discovered his passion for superbacteria e.coli during his studies which he gladly shared with us. In ten minutes, he taught us that e.coli does not only work as eeew on our meal, but showed how his group studies on making e.coli work for us, e.g. producing pharmaceuticals.</p><p>How? For example, recombinant DNA technology can be used to modify Escherichia coli to produce human insulin. Mogans works within a group of scientist that investigate the metabolic network in the Escherichia coli bacteria cell. Attempts to raise the copy number of plasmid DNA have been successful, and the goal now is to understand how the cell has accomplished this task. Proteomic studies have indicated an expected cellular adaptation that agrees with attempts to raise copy number in the literature; however, the thermodynamic implications of these results have been minimally explored. The goal of a cell remains controversial; one explanation is that the cellular metabolism seeks to maximize the entropy (minimize the Gibbs energy) of the system. Using GAMS as an optimization tool, it is explored if the reaction paths that have proven to be the most likely are also the paths that minimize the Gibbs energy of the system.</p><p>All in all, our science slam has been a great success which demanded a reenactment from a lot of sites.</p><p><b> Here are a few impressions from our event:</b></p><p> <img src=https://static.igem.org/mediawiki/2013/0/07/BonnScienceslam1.PNG> <br /><br /><i>Tobias Bald from experimental dermatology (University of Bonn) spoke about the Dark side of the immune system.</i></p><p><img src=https://static.igem.org/mediawiki/2013/d/dd/BonnScienceslam2.PNG><br /><br /><i>Prof. Dr. Herbert Dreiner gave us a review of Fukushima with full activity.</i></p><p><img src=https://static.igem.org/mediawiki/2013/0/0c/BonnScienceslam3.PNG><br /><br /><i>Our winner Mogan Ramesh shared his passion for superbacteria e.coli.</i></p><p><img src=https://static.igem.org/mediawiki/2013/2/24/BonnScienceslam4.PNG>< br /><br /><i>Full house at our Science Slam.</i></p><p><img src=https://static.igem.org/mediawiki/2013/d/d5/BonnScienceslam5.PNG><br /><br /><i>Our audience obviously enjoyed the show.</i></p>";

+

content.text = "<h1>Science Slam</h1><p>Our project aims to make versatile control of biological machines easily possible for everybody. During conception we wondered how our human practice work could represent that thought and decided to organize a science slam an event which makes science available to everybody!</p><p>We believe that science must not only stay in laboratories and lecture halls but reach lots of people out there. And without a doubt, the walls of the reputed ivory tower couldnt stand throughout the evening of our event!</p><p>Furthermore, the science slam provided a platform for our slammers to share the passion for their project with more than just their colleagues and thus help to popularize their field. In ten minutes, they presented their topic in a playful and inspirational manner and the audience decided which slammer has managed to get them carried away the most.</p><p>We managed not only to give floor to a large variety of fields by our slammers from physics, medicine, psychology, informatics, politics and engineering but to make them be heard by a public which was very mixed indeed.</p><p>As a prelude to the event, we acquainted the audience with synthetic biology and our project by showing what synthetic biology can do for each of us. Distributed questionnaires during the evening showed that -as hoped- our presentation helped to leave a positive mark on the field of synthetic biology in people's heads:</p><p>For example, 97% of the people, who stated they did not know about synthetic biology before have got a proper picture of it during our event. 86% said, they are now familiar with the iGEM competition and everybody stated, they found the idea to combine the presentation of our iGEM project with a science slam very good or good. </br> <p> <img src=https://static.igem.org/mediawiki/2013/d/df/Bonn_Science_Slam_1.png> </br> <p> <img src= https://static.igem.org/mediawiki/2013/f/f1/Bonn_Science_Slam_2.png> </br> <p> <img src=https://static.igem.org/mediawiki/2013/5/55/Bonn_Science_Slam_3.png></br> </p><p>The fact that our audience liked this topic had been reflected in our voting: Our representative for synthetic biology, Mogan Ramesh won the science slam. He originally comes from engineering but discovered his passion for superbacteria e.coli during his studies which he gladly shared with us. In ten minutes, he taught us that e.coli does not only work as eeew on our meal, but showed how his group studies on making e.coli work for us, e.g. producing pharmaceuticals.</p><p>How? For example, recombinant DNA technology can be used to modify Escherichia coli to produce human insulin. Mogans works within a group of scientist that investigate the metabolic network in the Escherichia coli bacteria cell. Attempts to raise the copy number of plasmid DNA have been successful, and the goal now is to understand how the cell has accomplished this task. Proteomic studies have indicated an expected cellular adaptation that agrees with attempts to raise copy number in the literature; however, the thermodynamic implications of these results have been minimally explored. The goal of a cell remains controversial; one explanation is that the cellular metabolism seeks to maximize the entropy (minimize the Gibbs energy) of the system. Using GAMS as an optimization tool, it is explored if the reaction paths that have proven to be the most likely are also the paths that minimize the Gibbs energy of the system.</p><p>All in all, our science slam has been a great success which demanded a reenactment from a lot of sites.</p><p><b> Here are a few impressions from our event:</b></p><p> <img src=https://static.igem.org/mediawiki/2013/0/07/BonnScienceslam1.PNG> <br /><br /><i>Tobias Bald from experimental dermatology (University of Bonn) spoke about the Dark side of the immune system.</i></p><p><img src=https://static.igem.org/mediawiki/2013/d/dd/BonnScienceslam2.PNG><br /><br /><i>Prof. Dr. Herbert Dreiner gave us a review of Fukushima with full activity.</i></p><p><img src=https://static.igem.org/mediawiki/2013/0/0c/BonnScienceslam3.PNG><br /><br /><i>Our winner Mogan Ramesh shared his passion for superbacteria e.coli.</i></p><p><img src=https://static.igem.org/mediawiki/2013/2/24/BonnScienceslam4.PNG><br/><br/><i>Full house at our Science Slam.</i></p><p><img src=https://static.igem.org/mediawiki/2013/d/d5/BonnScienceslam5.PNG><br/><br/><i>Our audience obviously enjoyed the show.</i></p>";

content.type = "Human Practice";

break;

Line 819:

Line 735:

case 107:

content.i = 107;

-

content.parents=[~~105~~];

+

content.parents=[100];

+

content.childs=[];

content.titleShort = "SynBio Day";

content.titleLong = "SynBio Day";

Line 830:

Line 747:

content.parents=[100];

content.childs=[];

-

content.titleShort = "~~School presentations~~";

+

content.titleShort = "Schools";

content.titleLong = "School presentations";

content.summary= "We gave lectures about synthetic biology and our project in schools in order to reach the younger people. ";

Line 840:

Line 757:

case 109:

content.i = 109;

-

content.parents=[~~105~~];

+

content.parents=[112];

-

content.childs=[~~112~~];

+

content.childs=[];

-

content.titleShort = "About ~~the comic~~";

+

content.titleShort = "Comic: About";

content.titleLong = "About the comic";

content.summary= "We introduced a hand drawn comic series called "LOV Wars" about Obi-Wan E. Coli and his adventures in the Galaxy of Petri. Basic concepts of synthetic biology are explained along the way.";

-

content.text="On the mission to find new and interesting means to bring across the concepts of synthetic biology, we introduced a comic series consisting of three episodes in the style of the well known Star Wars movies. The readers will find themselves in a world where Galaxies are petri dishes and all the characters are bacteria. Alongside the action-filled story we step by step introduce basic concepts of synthetic biology. The use of light sabers and laser guns also offered a great opportunity to embed our system of light-degradable proteins in the plot. At our presentations at schools and at our information booth it proved to be an ideal eye-catcher for passers-by and led to them wanting to know more about the subject.

+

content.text="On the mission to find new and interesting means to bring across the concepts of synthetic biology, we introduced a comic series consisting of three episodes in the style of the well known Star Wars movies. The readers will find themselves in a world where Galaxies are petri dishes and all the characters are bacteria. Alongside the action-filled story we step by step introduce basic concepts of synthetic biology. The use of light sabers and laser guns also offered a great opportunity to embed our system of light-degradable proteins in the plot. At our presentations at schools and at our information booth it proved to be an ideal eye-catcher for passers-by and led to them wanting to know more about the subject. During the episodes the reader accompanies the hero Obi-Wan E. Coli and his Padawan Plasmida on their journey through the Galaxy of Petri. They are fighting the villain Darth Cherry and his companions, the clones. But we don't want to spoil the story for you, just read the comic yourself below."; content.type="Human Practice";

-

During the episodes the reader accompanies the hero Obi-Wan E. Coli and his Padawan Plasmida on their journey through the Galaxy of Petri. They are fighting the villain Darth Cherry and his companions, the clones. But we ~~don’t~~ want to spoil the story for you, just read the comic yourself below.";

+

-

content.type="Human Practice";

+

break;

Line 854:

Line 769:

content.parents=[100];

content.childs=[];

-

content.titleShort = "~~About the~~ LOV-Wars ~~Shooter~~";

+

content.titleShort = "LOV-Wars Minigame</br>About";

content.titleLong = "About the LOV-Wars Shooter";

content.summary= "The thoughts and ideas behind our game.";

Line 875:

Line 790:

case 112:

content.i = 112;

-

content.parents = [100~~, 109~~];

+

content.parents = [100];

-

content.childs = [];

+

content.childs = [109];

content.titleShort = "Comic";

content.titleLong = "Comic – The adventures of Obi Wan E.Coli";

content.summary = "Read about the adventures of Obi Wan E.Coli";

-

content.text = "<img src=https://static.igem.org/mediawiki/2013/c/c1/Bonn_Comic_episode1_page1.jpg width=~~896px~~><img src=https://static.igem.org/mediawiki/2013/a/ad/Bonn_Comic_episode1_page2.jpg width=~~896px~~><img src=https://static.igem.org/mediawiki/2013/1/19/Bonn_Comic_episode1_page3.jpg width=~~896px~~><img src=https://static.igem.org/mediawiki/2013/8/80/Bonn_Comic_episode1_page4.jpg width=~~896px~~><img src=https://static.igem.org/mediawiki/2013/6/6c/Comic_EN_2.0_%281%29.jpg width=~~896px~~><img src=https://static.igem.org/mediawiki/2013/8/84/Comic_EN_2.0_%282%29.jpg width=~~896px~~><img src=https://static.igem.org/mediawiki/2013/9/97/Comic_EN_2.0_%283%29.jpg width=~~896px~~><img src=https://static.igem.org/mediawiki/2013/1/19/Comic_EN_2.0_%284%29.jpg width=~~896px~~><img src=https://static.igem.org/mediawiki/2013/8/8d/Bonn_Comic_EN_3.0_%281%29.jpg width=~~896px~~><img src=https://static.igem.org/mediawiki/2013/5/58/Bonn_Comic_EN_3.0_%282%29.jpg width=~~896px~~><img src=https://static.igem.org/mediawiki/2013/8/8c/Bonn_Comic_EN_3.0_%283%29.jpg width=~~896px~~><img src=https://static.igem.org/mediawiki/2013/b/b4/Bonn_Comic_EN_3.0_%284%29.jpg width=~~896px~~>";

+

content.text = "<img src=https://static.igem.org/mediawiki/2013/c/c1/Bonn_Comic_episode1_page1.jpg width=735px><img src=https://static.igem.org/mediawiki/2013/a/ad/Bonn_Comic_episode1_page2.jpg width=735px><img src=https://static.igem.org/mediawiki/2013/1/19/Bonn_Comic_episode1_page3.jpg width=735px><img src=https://static.igem.org/mediawiki/2013/8/80/Bonn_Comic_episode1_page4.jpg width=735px><img src=https://static.igem.org/mediawiki/2013/6/6c/Comic_EN_2.0_%281%29.jpg width=735px><img src=https://static.igem.org/mediawiki/2013/8/84/Comic_EN_2.0_%282%29.jpg width=735px><img src=https://static.igem.org/mediawiki/2013/9/97/Comic_EN_2.0_%283%29.jpg width=735px><img src=https://static.igem.org/mediawiki/2013/1/19/Comic_EN_2.0_%284%29.jpg width=735px><img src=https://static.igem.org/mediawiki/2013/8/8d/Bonn_Comic_EN_3.0_%281%29.jpg width=735px><img src=https://static.igem.org/mediawiki/2013/5/58/Bonn_Comic_EN_3.0_%282%29.jpg width=735px><img src=https://static.igem.org/mediawiki/2013/8/8c/Bonn_Comic_EN_3.0_%283%29.jpg width=735px><img src=https://static.igem.org/mediawiki/2013/b/b4/Bonn_Comic_EN_3.0_%284%29.jpg width=735px>";

content.type = "Human Practice";

break;

Line 889:

Line 804:

content.i = 130;

content.parents=[100];

-

content.childs=[131,133~~,134,135~~,136,137,138];

+

content.childs=[131,133,136,137,138];

content.titleShort = "About us";

content.titleLong = "About us";

content.summary= "The Team - Presentation of all team members";

-

content.text= "<div><div style='float:right;display:inline; '><img src='https://static.igem.org/mediawiki/2013/a/a4/Bonn_sponsors_uniBonn.png' class='bottom-sponsor' height='50px' id='sponsor-unibonn' style='margin-right:10px;'><img src='https://static.igem.org/mediawiki/2013/7/79/Bonn_sponsors_limes.gif' class='bottom-sponsor' height='50px' id='sponsor-limes'></div><div class='subpage-head-link'><a href='javascript:history.back();'>Back</a></br> Category: Team</div></html>= About Us =<html><div class='subpage-text'><div align='center'><div width:870px; height:485px; id='aboutus-group' style='position:relative'><img src='https://static.igem.org/mediawiki/2013/e/e9/Teammod.jpg' width='870px' style='z-index:1;position:relative;'><div id='aboutus-group-kristina' onmouseover=showMemberDetails('Kristina') onmouseout=hideMemberDetails() style='position:absolute;top:160px;left:57px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-dustin' onmouseover=showMemberDetails('Dustin') onmouseout=hideMemberDetails() style='position:absolute;top:153px;left:130px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-marc' onmouseover=showMemberDetails('Marc') onmouseout=hideMemberDetails() style='position:absolute;top:275px;left:124px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-niklas' onmouseover=showMemberDetails('Niklas') onmouseout=hideMemberDetails() style='position:absolute;top:139px;left:174px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-ben' onmouseover=showMemberDetails('Ben') onmouseout=hideMemberDetails() style='position:absolute;top:158px;left:197px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-annika' onmouseover=showMemberDetails('Annika') onmouseout=hideMemberDetails() style='position:absolute;top:278px;left:220px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-matthias' onmouseover=showMemberDetails('Matthias') onmouseout=hideMemberDetails() style='position:absolute;top:153px;left:252px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-max' onmouseover=showMemberDetails('Max') onmouseout=hideMemberDetails() style='position:absolute;top:159px;left:282px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-maria' onmouseover=showMemberDetails('Maria') onmouseout=hideMemberDetails() style='position:absolute;top:273px;left:289px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-sadrija' onmouseover=showMemberDetails('Sadrija') onmouseout=hideMemberDetails() style='position:absolute;top:160px;left:358px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-franzi' onmouseover=showMemberDetails('Franzi') onmouseout=hideMemberDetails() style='position:absolute;top:277px;left:355px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-cori' onmouseover=showMemberDetails('Cori') onmouseout=hideMemberDetails() style='position:absolute;top:172px;left:420px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-pauline' onmouseover=showMemberDetails('Pauline') onmouseout=hideMemberDetails() style='position:absolute;top:282px;left:421px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-olli' onmouseover=showMemberDetails('Olli') onmouseout=hideMemberDetails() style='position:absolute;top:163px;left:472px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-nina' onmouseover=showMemberDetails('Nina') onmouseout=hideMemberDetails() style='position:absolute;top:277px;left:489px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-miriam' onmouseover=showMemberDetails('Miriam') onmouseout=hideMemberDetails() style='position:absolute;top:163px;left:506px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-thomas' onmouseover=showMemberDetails('Thomas') onmouseout=hideMemberDetails() style='position:absolute;top:140px;left:535px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-katharina' onmouseover=showMemberDetails('Katharina') onmouseout=hideMemberDetails() style='position:absolute;top:276px;left:557px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-philipp' onmouseover=showMemberDetails('Philipp') onmouseout=hideMemberDetails() style='position:absolute;top:147px;left:591px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-martina' onmouseover=showMemberDetails('MartinaB') onmouseout=hideMemberDetails() style='position:absolute;top:162px;left:622px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-sebastian' onmouseover=showMemberDetails('Sebastian') onmouseout=hideMemberDetails() style='position:absolute;top:153px;left:664px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-annikag' onmouseover=showMemberDetails('AnnikaG') onmouseout=hideMemberDetails() style='position:absolute;top:150px;left:703px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-jan' onmouseover=showMemberDetails('Jan') onmouseout=hideMemberDetails() style='position:absolute;top:136px;left:769px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-details' style='background-color:black;opacity:0.8;position:absolute;height:80px;width=870px;top:0px;left:0px;right:-24px;z-index:2;'><div id='aboutus-group-details-name' style='color:white'></div><div id='aboutus-group-details-course' style='color:white'></div><div id='aboutus-group-details-semester' style='color:white'></div><div id='aboutus-group-details-tasks' style='color:white'></div></div><div id='aboutus-group-move' style='background-color:black;opacity:0.8;position:absolute;height:20px;width=870px;top:0px;left:0px;right:-24px;z-index:2;'><div id='aboutus-group-about' style='color:white'>Hover over faces to show group member details.</div></div></div></div></div><div id='team-members'><div class='team-members-row'><div class='team-member'><div class='team-member-name'>Thomas Berger</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork, jamboree presentation, school presentations</div></div><div class='team-member'><div class='team-member-name'>Sadrija Cukoski</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>4th semester</div><div class='job'>science slam</div></div><div class='team-member'><div class='team-member-name'>Dustin Dankelmann</div><div class='team-member-field'>Bachelor Biology</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork, financing</div></div></div><div class='team-members-row'><div class='team-member'><div class='team-member-name'>Katharina Düker</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>4th semester</div><div class='job'>startup advisory</div></div><div class='team-member'><div class='team-member-name'>Maria Gädeke</div><div class='team-member-field'>Bachelor Biology</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork</div></div><div class='team-member'><div class='team-member-name'>Jan Hansen</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork, webdesign, lov-wars shooter, design, comic, school presentations, pictures</div></div></div><div class='team-members-row'><div class='team-member'><div class='team-member-name'>Kristina Klein</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork</div></div><div class='team-member'><div class='team-member-name'>Matthias Klumpp</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>4th semester</div><div class='job'>webmaster</div></div><div class='team-member'><div class='team-member-name'>Franziska Kohl</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>4th semester</div><div class='job'>survey and evaluation, school presentations</div></div></div><div class='team-members-row'><div class='team-member'><div class='team-member-name'>Sebastian Martin</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>4th semester</div><div class='job'>safety</div></div><div class='team-member'><div class='team-member-name'>Miriam Melake</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>2nd semester</div><div class='job'>science slam</div></div><div class='team-member'><div class='team-member-name'>Nina Offermann</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork, comic, school presentations</div></div></div><div class='team-members-row'><div class='team-member'><div class='team-member-name'>Oliver Rippel</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork, school presentations</div></div><div class='team-member'><div class='team-member-name'>Philipp Sander</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>4th semester</div><div class='job'>startup advisory</div></div><div class='team-member'><div class='team-member-name'>Florian Schäfer</div><div class='team-member-field'>Bachelor Mathematics</div><div class='team-member-sem'>6th semester</div><div class='job'>modelling</div></div></div><div class='team-members-row'><div class='team-member'><div class='team-member-name'>Max Schelski</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>4th semester</div><div class='job'>management, labwork, jamboree presentation, webdesign, school presentations</div></div><div class='team-member'><div class='team-member-name'>Niklas Schmacke</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>4th semester</div><div class='job'>labwork, jamboree presentation, webdesign, design</div></div><div class='team-member'><div class='team-member-name'>Corinna Schmalohr</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork, comic, school presentation, design</div></div></div><div class='team-members-row'><div class='team-member'><div class='team-member-name'>Annika Schneider</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork, jamboree presentation, meetings, financing</div></div><div class='team-member'><div class='team-member-name'>Marc Schulte</div><div class='team-member-field'>Master Molecular Biotechnology</div><div class='team-member-sem'>4th semester</div><div class='job'>labwork, design</div></div><div class='team-member'><div class='team-member-name'>Benjamin Syllwasschy</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork, school presentations</div></div></div><div class='team-members-row'><div class='team-member'><div class='team-member-name'>Pauline Ulmke</div><div class='team-member-field'>Bachelor Applied Biology</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork, school presentation</div></div><div class='team-member'><div class='team-member-name'>Leonie von Berlin</div><div class='team-member-field'>Bachelor Biology</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork</div></div></div></div></div></div></div></div>";

+

content.text= "<div><div class='subpage-text'><div align='center'><div width:870px; height:485px; id='aboutus-group' style='position:relative;left:-90px'><img src='https://static.igem.org/mediawiki/2013/e/e9/Teammod.jpg' width='870px' style='z-index:1;position:relative;'><div id='aboutus-group-kristina' onmouseover=showMemberDetails('Kristina') onmouseout=hideMemberDetails() style='position:absolute;top:160px;left:57px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-dustin' onmouseover=showMemberDetails('Dustin') onmouseout=hideMemberDetails() style='position:absolute;top:153px;left:130px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-marc' onmouseover=showMemberDetails('Marc') onmouseout=hideMemberDetails() style='position:absolute;top:275px;left:124px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-niklas' onmouseover=showMemberDetails('Niklas') onmouseout=hideMemberDetails() style='position:absolute;top:139px;left:174px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-ben' onmouseover=showMemberDetails('Ben') onmouseout=hideMemberDetails() style='position:absolute;top:158px;left:197px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-annika' onmouseover=showMemberDetails('Annika') onmouseout=hideMemberDetails() style='position:absolute;top:278px;left:220px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-matthias' onmouseover=showMemberDetails('Matthias') onmouseout=hideMemberDetails() style='position:absolute;top:153px;left:252px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-max' onmouseover=showMemberDetails('Max') onmouseout=hideMemberDetails() style='position:absolute;top:159px;left:282px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-maria' onmouseover=showMemberDetails('Maria') onmouseout=hideMemberDetails() style='position:absolute;top:273px;left:289px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-sadrija' onmouseover=showMemberDetails('Sadrija') onmouseout=hideMemberDetails() style='position:absolute;top:160px;left:358px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-franzi' onmouseover=showMemberDetails('Franzi') onmouseout=hideMemberDetails() style='position:absolute;top:277px;left:355px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-cori' onmouseover=showMemberDetails('Cori') onmouseout=hideMemberDetails() style='position:absolute;top:172px;left:420px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-pauline' onmouseover=showMemberDetails('Pauline') onmouseout=hideMemberDetails() style='position:absolute;top:282px;left:421px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-olli' onmouseover=showMemberDetails('Olli') onmouseout=hideMemberDetails() style='position:absolute;top:163px;left:472px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-nina' onmouseover=showMemberDetails('Nina') onmouseout=hideMemberDetails() style='position:absolute;top:277px;left:489px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-miriam' onmouseover=showMemberDetails('Miriam') onmouseout=hideMemberDetails() style='position:absolute;top:163px;left:506px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-thomas' onmouseover=showMemberDetails('Thomas') onmouseout=hideMemberDetails() style='position:absolute;top:140px;left:535px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-katharina' onmouseover=showMemberDetails('Katharina') onmouseout=hideMemberDetails() style='position:absolute;top:276px;left:557px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-philipp' onmouseover=showMemberDetails('Philipp') onmouseout=hideMemberDetails() style='position:absolute;top:147px;left:591px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-martina' onmouseover=showMemberDetails('MartinaB') onmouseout=hideMemberDetails() style='position:absolute;top:162px;left:622px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-sebastian' onmouseover=showMemberDetails('Sebastian') onmouseout=hideMemberDetails() style='position:absolute;top:153px;left:664px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-annikag' onmouseover=showMemberDetails('AnnikaG') onmouseout=hideMemberDetails() style='position:absolute;top:150px;left:703px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-jan' onmouseover=showMemberDetails('Jan') onmouseout=hideMemberDetails() style='position:absolute;top:136px;left:769px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-details' style='background-color:black;opacity:0.8;position:absolute;height:80px;width=870px;top:0px;left:0px;right:-24px;z-index:2;'><div id='aboutus-group-details-name' style='color:white'></div><div id='aboutus-group-details-course' style='color:white'></div><div id='aboutus-group-details-semester' style='color:white'></div><div id='aboutus-group-details-tasks' style='color:white'></div></div><div id='aboutus-group-move' style='background-color:black;opacity:0.8;position:absolute;height:20px;width=870px;top:0px;left:0px;right:-24px;z-index:2;'><div id='aboutus-group-about' style='color:white'>Hover over faces to show group member details.</div></div></div></div></div><div id='team-members'><div class='team-members-row'><div class='team-member'><div class='team-member-name'>Thomas Berger</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork, jamboree presentation, school presentations</div></div><div class='team-member'><div class='team-member-name'>Sadrija Cukoski</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>4th semester</div><div class='job'>science slam</div></div><div class='team-member'><div class='team-member-name'>Dustin Dankelmann</div><div class='team-member-field'>Bachelor Biology</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork, financing</div></div></div><div class='team-members-row'><div class='team-member'><div class='team-member-name'>Katharina Düker</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>4th semester</div><div class='job'>startup advice, skills course, school presentation</div></div><div class='team-member'><div class='team-member-name'>Maria Gädeke</div><div class='team-member-field'>Bachelor Biology</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork</div></div><div class='team-member'><div class='team-member-name'>Jan Hansen</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork, webdesign, lov-wars shooter, design, comic, school presentations, pictures</div></div></div><div class='team-members-row'><div class='team-member'><div class='team-member-name'>Kristina Klein</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork</div></div><div class='team-member'><div class='team-member-name'>Matthias Klumpp</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>4th semester</div><div class='job'>webmaster</div></div><div class='team-member'><div class='team-member-name'>Franziska Kohl</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>4th semester</div><div class='job'>survey and evaluation, school presentations</div></div></div><div class='team-members-row'><div class='team-member'><div class='team-member-name'>Sebastian Martin</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>4th semester</div><div class='job'>safety</div></div><div class='team-member'><div class='team-member-name'>Miriam Melake</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>2nd semester</div><div class='job'>science slam</div></div><div class='team-member'><div class='team-member-name'>Nina Offermann</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork, comic, school presentations</div></div></div><div class='team-members-row'><div class='team-member'><div class='team-member-name'>Oliver Rippel</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork, school presentations</div></div><div class='team-member'><div class='team-member-name'>Philipp Sander</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>4th semester</div><div class='job'>startup advice, financing, skills course, school presentation</div></div><div class='team-member'><div class='team-member-name'>Florian Schäfer</div><div class='team-member-field'>Bachelor Mathematics</div><div class='team-member-sem'>6th semester</div><div class='job'>modelling</div></div></div><div class='team-members-row'><div class='team-member'><div class='team-member-name'>Max Schelski</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>4th semester</div><div class='job'>management, labwork, jamboree presentation, jamboree poster, webdesign, school presentations, skills course</div></div><div class='team-member'><div class='team-member-name'>Niklas Schmacke</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>4th semester</div><div class='job'>labwork, jamboree presentation, jamboree poster webdesign, design</div></div><div class='team-member'><div class='team-member-name'>Corinna Schmalohr</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork, comic, school presentation, design</div></div></div><div class='team-members-row'><div class='team-member'><div class='team-member-name'>Annika Schneider</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork, jamboree presentation, meetings, financing</div></div><div class='team-member'><div class='team-member-name'>Marc Schulte</div><div class='team-member-field'>Master Molecular Biotechnology</div><div class='team-member-sem'>4th semester</div><div class='job'>labwork, design</div></div><div class='team-member'><div class='team-member-name'>Benjamin Syllwasschy</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork, school presentations</div></div></div><div class='team-members-row'><div class='team-member'><div class='team-member-name'>Pauline Ulmke</div><div class='team-member-field'>Bachelor Applied Biology</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork, school presentation</div></div><div class='team-member'><div class='team-member-name'>Leonie von Berlin</div><div class='team-member-field'>Bachelor Biology</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork</div></div></div></div></div></div>";

content.type="Team";

break;

Line 915:

Line 830:

content.titleLong = "Sponsors";

content.summary= "Many sponsors made our work possible.";

-

content.text= "<div class=subpage-text> <table class=subpage-sponsors> <tr class=subpage-sponsor style=border-style:solid;border-width:5px;border-color:grey;> <td class=subpage-sponsor> <img src=https://static.igem.org/mediawiki/2013/3/36/Bonn_sponsor_promega.png class=bottom-sponsor width=300px id=sponsor-limes> </td> <td> <h2>Promega</h2> </br> Promega is one of the five biggest worldwide acting Life Science Research company. It was founded in Madison, WI (USA) and produces products and system solutions for gen-, protein- and cell-analysis. With the help of these products biological systems can be explored easily. Promega-products can be used in basic research, development of medicaments, molecular diagnostic and identification of human genetic constitution. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/3/36/Bonn_sponsors_eppendorf.jpg class=bottom-sponsor width=300px id=sponsor-eppendorf> </td> <td> <h2>Eppendorf</h2> </br> Eppendorf is a biotechnical company that develops, produces and sells systems for life science research for laboratories all over the world. The assortment of goods contains pipettes, dispensers, centrifuges, reaction tubes and pipette tips. Moreover Eppendorf offers instruments and systems to manipulate cells, automated machines for Liquid Handling and for DNA- Amplification, as well as Biochips. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/5/5a/Bonn_sponsor_IKA.png class=bottom-sponsor width=300px id=sponsor-ika> </td> <td><h2> IKA </h2> </br> In 1910 the companys history of IKA began, China had not been invented yet Peoples Republic and the word globalization. Today, the IKA group about 800 employees at eight locations on four continents and is pleased with clients such as BASF, Bayer and Procter & Gamble. In most product groups, we are sovereign world market leader and a symbol of development and growth. Or as our new slogan: "IKA - Designed to work perfectly." </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/b/bf/Bonn_sponsors_Ella.jpg class=bottom-sponsor width=300px'id=sponsor-ella> </td> <td> <h2>Ella Biotech</h2> </br> ELLA is an independent, privately owned company founded in October 2004. ELLA offers creative services for the production of oligonucleotides driven by the goal of continually improving our production strategies. ELLA offers tangible advantages to its customers and partners through its validated technology platform, its experienced interdisciplinary team, and its resolute attitude towards the highest quality in our products. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/c/c0/Bonn_sponsor_genscript.jpg class=bottom-sponsor width=370px id=sponsor-genscript> </td> <td> <h2> Gen Script </h2> </br> GenScript is a leading biology CRO focusing exclusively on early drug discovery and development services. Built on our assembly-line mode, one-stop solution, continuous improvement, and stringent IP protection, GenScript provides a comprehensive portfolio of services that include Bio-Reagent, Bio-Assay, Lead Optimization, and Antibody Drug Development which can be effectively integrated into your value chain and your operations. We strive with competence and confidence to meet your demand for developing pre-clinical drug candidates time-efficiently and cost-effectively. With track performance record, GenScript is your ideal and reliable innovation partner in drug discovery. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/1/13/Bonn_sponsor_idt.jpg class=bottom-sponsor width=300px id=sponsor-idt> </td> <td><h2> IDT </h2> </br> Genetically engineered vaccines and pharmaceuticals for the global marketplace. The IDT Biologicals is an innovative medium-sized company that is involved in more than 90 years with its products and services to the health maintenance of humans and animals. Genetically engineered vaccines and pharmaceuticals for the domestic and international market are manufactured. In fiscal year 2012, the IDT biologics had a turnover of around 151 million euros. In the IDT biologics around 1,100 people are currently employed. IDT Biologics is an independent company of the Klocke Group, which developed and implemented at five production sites innovative packaging solutions for the pharmaceutical, cosmetic, food and chemical-technical industry.</br> Decades of research and development of vaccines</br> Since the beginning of the IDT successfully fought the various pathogens in animals with complex vaccine development from the laboratory to production and sales in one location. From research and development to manufacturing and testing and approval, national and international marketing of the range of tasks. The IDT Animal Health operates its own modern research complex for the development of animal vaccines.</br> Integrated biopharmaceutical services. </br> Since its founding in 1921, the IDT has biologics developed into a center for the pharmaceutical and biotechnology with the divisions Animal Health, human vaccines and pharmaceuticals. More than 250 million euros have been invested since privatization in 1993 in the continuous expansion of an integrated biopharmaceutical site and thus not only created excellent conditions of production, but also highly modern workplaces. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/3/3a/Bonn_sponsor_neb.jpg class=bottom-sponsor width=300px id=sponsor-neb> </td> <td> <h2> New England Biolabs </h2> </br> Never before in the history of science, the demands on the molecular biology industry have been as high as today: The researcher asks for the absolute best and most reliable products - there is no room for compromise. New England Biolabs fulfills this requirement. For 35 years we have been a leader in the development and production of enzymes for molecular biology and other reagents in the "life sciences" such as for proteomics and drug discovery. Our expertise in enzyme technology based on our strategic cloning and expression of DNA-Restriktions-/Modifikationssystemen program. So we have for years set the standards in terms of quality and price. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/3/34/Bonn_sponsors_MN.png class=bottom-sponsor width=300px id=sponsor-mn> </td> <td> <h2> Macherey und Nagel </h2> </br> MACHEREY-NAGEL is a family-run concern in the fourth generation. The comprehensive portfolio includes the areas of filtration, rapid tests, water analysis, chromatography and bioanalysis. MACHEREY-NAGEL employs more than 470 highly skilled employees in sales, production as well as research and development, including 10% post-doctoral researchers. They all guarantee an exceptional service. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/3/37/Bonn_Sponsor_roche.jpg class=bottom-sponsor width=300px id=sponsor-roche> </td> <td> <h2> Roche </h2> </br> Roche Headquartered in Basel, Switzerland, is a leading research-focused healthcare company with the pharmaceuticals and diagnostics businesses. As the worlds largest biotech company developing clinically differentiated medicines in oncology, virology, inflammation, metabolism and central nervous system. Roche, a pioneer in diabetes management, is also the world leader in in-vitro diagnostics, tissue-based cancer diagnostics. Medicines and diagnostic tools that enable tangible improvements in the health, quality of life and survival of patients is the strategic goal of personalized medicine from Roche. This concept is based on new molecular insights and molecular diagnostic tests that allow a more precise tuning of therapy and better control of the disease. Therapies are tailored to patient groups that have similarities in their disease. The only way to improve the efficacy of drugs targeted and maintain quality of life. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/d/d2/Bonn_sponsor_geneious.gif class=bottom-sponsor width=300px id=sponsor-geneious> </td> <td> <h2> Geneious </h2> </br> In Good Company</br> First released in 2005, Geneious is one the worlds leading bioinformatics software platforms, used by over 2500 universities and institutes and commercial companies in more than 65 countries. Geneious is used by all 20 of the top 20 Universities globally (Times Higher Education, 2012) and by seven of the 10 largest pharmaceutical companies.</br> Dedicated to excellence</br> Our software has won a number of prestigious awards, including the Computerworld Excellence Awards from Innovative Use of ICT and the United Nations World Summer Awards and Winner in the e-Science Category in 2007, the Recruit IT Innovative Software Product Award at the PriceWaterhouseCoopers Hi-Tech Awards in 2009 and a Global Finalist in the IT and Informatics category at the Bio-IT World Awards in Boston in 2009. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/5/55/Bonn_Sponsor_vwr.jpg class=bottom-sponsor width=300px id=sponsor-vwr> </td> <td> <h2> VWR </h2> </br> VWR is science for the advancement of the worlds most important research through the distribution of a wide range of products and services to a variety of important companies in the pharmaceutical, biotechnology, and healthcare industries as well as government agencies, universities and schools. We offer our customers all the resources they need to be successful, ie an extensive range of the best products in the areas of chemicals, furniture, appliances, instruments, apparel and consumables, from a wide variety of leaders in the field of science manufacturers. With 160 years of experience in this industry, VWR further supports its customers through a combination of strength, vision, innovation and a well-established distribution network that reaches thousands of specialized labs and facilities across the planet. VWR is not just a product supplier - it keeps the most important research in the world in motion. VWRs expertise in the areas of supply chain and logistics services enables customers to fully concentrate on their areas of expertise. Of the management of procurement processes to the integration of supply chains: VWR helps specialized research facilities and laboratories to work with maximum efficiency. VWR has over 8,000 employees in 30 countries with direct offices throughout the world working to streamline the way, as researchers from North America, Europe and the Asia-Pacific region supply and maintain their labs. In addition, VWR further supports its customers by providing onsite services, storeroom management, product procurement, supply chain systems integration and technical services. We are expanding our global presence and adhere to the principle that customers benefit from the availability and expertise of our local sales teams.</br> In todays economy VWR helps its customers to focus on increasing productivity and reducing costs and optimizing procurement processes. </br> Headquartered in Radnor, PA (USA), earned VWR International, LLC, in 2012 global sales of more than 4.1 billion U.S. dollars. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/0/0b/Bonn_Sponsors_Roth.png class=bottom-sponsor width=170px id=sponsor-roth> </td> <td> <h2> ROTH – A COMPANY WITH TRADITION </h2> </br> 1879 </br> Carl ROTH founded in Karlsruhe, a "material, Colonial and dye business and Droguerie".</br>1899</br> The first sales and mail order catalog is published.</br> 1956</br> Publication of the first issue of the publication a "Rarea " Natural Products</br> 1990</br> First ROTH general catalog. Complete with the areas of laboratory, life sciences and chemicals in one.</br> 2005</br> Completion of a modern establishment for the production and storage of laboratory chemicals and reagents in Karlsruhe Rhine port area. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/7/79/Bonn_starlab.jpg class=bottom-sponsor width=300px id=sponsor-starlab> </td> <td> <h2> Starlab </h2> </br> STARLAB is a company specializing in liquid handling technology group. With subsidiaries in Germany, France, Britain and Italy is available in the direct sales an extensive range of products available. Plus, you get our products to many countries around the world via our international trading partners. Our success is based on many years of experience in manufacturing and marketing of liquid handling disposable products - with TipOne we have established ourselves as a leading supplier of pipette tips systems worldwide. This quality, price and service come first. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/b/bf/Bonn_caesar.gif class=bottom-sponsor width=200px id=sponsor-casar> </td> <td> <h2> Caesar </h2> </br> The center of advanced european studies and research (caesar) is an institute of the Max Planck Society, which is located at the boundaries between neuroscience, cell biology and biophysics. The focus of the research is the particular cellular and neural signal processing. </br> Caesar works with modern photonic, molecular biological, chemical and micro-technological methods. The focus of kinetic, spectroscopic and microscopic methods are research and control of cellular activity. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/6/6e/Bonn_Eurofins.png class=bottom-sponsor width=300px id=sponsor-eurofins> </td> <td> <h2> Eurofins MWG operon </h2> </br> Discover our fascination about the world of the four bases.</br> We are fascinated about the power of DNA and how it is incorporated in everything we do, work and live. Being passionate about our strong customer orientation, our service and our quality standards, we continuously challenge ourselves to stay ahead and remain one of the leading genomics service providers worldwide. Eurofins MWG Operon is globally known for its innovative and customised technologies in the life science industries and academic research institutions. With the combined power of an international network of Eurofins companies in the field of genomic services, forensics, agroscience, pharmaceutical, environmental, food and feed testing, we have established an outstanding team of experts and broad range of technologies. This unique constellation underlines our approach to offer best practise solutions and versatile concepts for our clients - academic institutions and large interdisciplinary operating companies of the world. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/1/18/Bonn_ThermoFisher.jpg class=bottom-sponsor width=300px id=sponsor-thermofisher> </td> <td> <h2> Thermo Fisher </h2> </br> Thermo Fisher Scientific Inc. (NYSE: TMO) is the world leader in serving science. Our mission is to enable our customers to make the world healthier, cleaner and safer. With revenues of $13 billion, we have approximately 39,000 employees and serve customers within pharmaceutical and biotech companies, hospitals and clinical diagnostic labs, universities, research institutions and government agencies, as well as in environmental and process control industries. We create value for our key stakeholders through three premier brands, Thermo Scientific, Fisher Scientific and Unity<sup> TM </sup> Lab Services, which offer a unique combination of innovative technologies, convenient purchasing options and a single solution for laboratory operations management. Our products and services help our customers solve complex analytical challenges, improve patient diagnostics and increase laboratory productivity. </td></tr></table> </div> </div> </div> </div> </div> ";

+

content.text= "<div class=subpage-text> <table class=subpage-sponsors style=margin-left:-50px;background-color:transparent><tr><td colspan=2><h2>Our Partners</h2></td></tr><tr class=subpage-sponsor style=border-style:solid;border-width:5px;border-color:grey;> <td colspan=2 class=subpage-sponsor> <h2>BIO.NRW Cluster Biotechnology North Rhine-Westphalia</h2> <img src=https://static.igem.org/mediawiki/2013/9/90/Bonn_sponsor_BioNRW.png class=bottom-sponsor width=500px id=sponsor-limes style=margin-bottom:-50px></td><tr><td colspan=2></br>North Rhine-Westphalia is situated at Europe’s geographic and economic center. It is the largest of Germany’s 16 federal states, and the number one exporter. If classified as an independent exporting nation, NRW would rank 18th in the world, on a par with the Netherlands.</br>North Rhine-Westphalia’s state government has established a number of technology clusters to systematically improve NRW’s strengths and talents in established industries and up-and-coming fields like biotechnology. Goal of the “ExcellenceNRW” cluster strategy is to create a favourable climate for innovation, as that is the best way to sustain the competitive edge and stimulate growth and employment in the companies that call the state their home.North Rhine-Westphalia’s biotechnology cluster BIO.NRW is a central catalyst for the sustainable development of the state’s biotech sector. It activates cooperation between business, research, investors and policy-makers. The cluster also promotes the strengths and achievements of biotechnology in the state.</br>To support young as well as already established biotech companies, BIO.NRW offers the following core competencies:</br>– Individual matchmaking for collaborations and partners</br>– Overview on all up-to-date R&D activities in industry and academia</br>– Profound knowledge on financing possibilities</br>– International promotion and marketing for NRW as biotech location</br>– Direct contacts to decision makers</br><h3>Our services include</h3><bold>Technology Transfer</bold></br>Tech transfer support is a key contribution from BIO.NRW. We organize events, working platforms and meetings to promote the dialogue between all stakeholders in the field of biotechnology and to encourage cooperation.</br><bold>Biotech Business & Sciences</bold></br>BIO.NRW compiles comprehensive and current online databases of the academic institutions and companies active in the life sciences in NRW. Free to access and easy-to-use, these resources are valuable tools for identifying prospective business partners. More information on <a href=http://www.bio.nrw.de>www.bio.nrw.de</a></br><bold>Fairs, Exhibitions and Conferences</bold></br>Companies and academic institutions can generate awareness of their activities locally, nationally and internationally by being a part of the BIO.NRW common stands on fairs, exhibitions and conferences. BIO.NRW also organizes a series of workshops and symposia, called BIO.NRW.academy.</br><bold>Support of Young Professionals</bold></br>BIO.NRW takes a special interest in supporting young professionals in biotechnology. For example, we organize conventions where graduates meet representatives from industry and academic science. The ‘Business Angel Network – BIO.NRW’ helps financing and funding biotech start-ups. In addition, a forum that brings together investment institutions, private investors and business angels and developers provides information about the current NRW biotech scene. These meetings are a valuable opportunity for start-up companies to receive coaching and financing.</br></br>To learn more about the Cluster BIO.NRW and to stay informed about the latest biotech developments in NRW please visit <a href=http://www.bio.nrw.de>www.bio.nrw.de</a>.</td> </tr><tr><td colspan=2><h2>Our Sponsors</h2></td></tr><tr class=subpage-sponsor style=border-style:solid;border-width:5px;border-color:grey;> <td class=subpage-sponsor> <img src=https://static.igem.org/mediawiki/2013/3/36/Bonn_sponsor_promega.png class=bottom-sponsor width=250px id=sponsor-limes> </td> <td> <h2>Promega</h2> </br> Promega is one of the five biggest worldwide acting Life Science Research company. It was founded in Madison, WI (USA) and produces products and system solutions for gen-, protein- and cell-analysis. With the help of these products biological systems can be explored easily. Promega-products can be used in basic research, development of medicaments, molecular diagnostic and identification of human genetic constitution. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/3/36/Bonn_sponsors_eppendorf.jpg class=bottom-sponsor width=250px id=sponsor-eppendorf> </td> <td> <h2>Eppendorf</h2> </br> Eppendorf is a biotechnical company that develops, produces and sells systems for life science research for laboratories all over the world. The assortment of goods contains pipettes, dispensers, centrifuges, reaction tubes and pipette tips. Moreover Eppendorf offers instruments and systems to manipulate cells, automated machines for Liquid Handling and for DNA- Amplification, as well as Biochips. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/5/5a/Bonn_sponsor_IKA.png class=bottom-sponsor width=250px id=sponsor-ika> </td> <td><h2> IKA </h2> </br> In 1910 the companys history of IKA began, China had not been invented yet Peoples Republic and the word globalization. Today, the IKA group about 800 employees at eight locations on four continents and is pleased with clients such as BASF, Bayer and Procter & Gamble. In most product groups, we are sovereign world market leader and a symbol of development and growth. Or as our new slogan: "IKA - Designed to work perfectly." </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/b/bf/Bonn_sponsors_Ella.jpg class=bottom-sponsor width=250px'id=sponsor-ella> </td> <td> <h2>Ella Biotech</h2> </br> ELLA is an independent, privately owned company founded in October 2004. ELLA offers creative services for the production of oligonucleotides driven by the goal of continually improving our production strategies. ELLA offers tangible advantages to its customers and partners through its validated technology platform, its experienced interdisciplinary team, and its resolute attitude towards the highest quality in our products. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/c/c0/Bonn_sponsor_genscript.jpg class=bottom-sponsor width=370px id=sponsor-genscript> </td> <td> <h2> Gen Script </h2> </br> GenScript is a leading biology CRO focusing exclusively on early drug discovery and development services. Built on our assembly-line mode, one-stop solution, continuous improvement, and stringent IP protection, GenScript provides a comprehensive portfolio of services that include Bio-Reagent, Bio-Assay, Lead Optimization, and Antibody Drug Development which can be effectively integrated into your value chain and your operations. We strive with competence and confidence to meet your demand for developing pre-clinical drug candidates time-efficiently and cost-effectively. With track performance record, GenScript is your ideal and reliable innovation partner in drug discovery. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/1/13/Bonn_sponsor_idt.jpg class=bottom-sponsor width=250px id=sponsor-idt> </td> <td><h2> IDT </h2> </br> Genetically engineered vaccines and pharmaceuticals for the global marketplace. The IDT Biologicals is an innovative medium-sized company that is involved in more than 90 years with its products and services to the health maintenance of humans and animals. Genetically engineered vaccines and pharmaceuticals for the domestic and international market are manufactured. In fiscal year 2012, the IDT biologics had a turnover of around 151 million euros. In the IDT biologics around 1,100 people are currently employed. IDT Biologics is an independent company of the Klocke Group, which developed and implemented at five production sites innovative packaging solutions for the pharmaceutical, cosmetic, food and chemical-technical industry.</br> Decades of research and development of vaccines</br> Since the beginning of the IDT successfully fought the various pathogens in animals with complex vaccine development from the laboratory to production and sales in one location. From research and development to manufacturing and testing and approval, national and international marketing of the range of tasks. The IDT Animal Health operates its own modern research complex for the development of animal vaccines.</br> Integrated biopharmaceutical services. </br> Since its founding in 1921, the IDT has biologics developed into a center for the pharmaceutical and biotechnology with the divisions Animal Health, human vaccines and pharmaceuticals. More than 250 million euros have been invested since privatization in 1993 in the continuous expansion of an integrated biopharmaceutical site and thus not only created excellent conditions of production, but also highly modern workplaces. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/3/3a/Bonn_sponsor_neb.jpg class=bottom-sponsor width=250px id=sponsor-neb> </td> <td> <h2> New England Biolabs </h2> </br> Never before in the history of science, the demands on the molecular biology industry have been as high as today: The researcher asks for the absolute best and most reliable products - there is no room for compromise. New England Biolabs fulfills this requirement. For 35 years we have been a leader in the development and production of enzymes for molecular biology and other reagents in the "life sciences" such as for proteomics and drug discovery. Our expertise in enzyme technology based on our strategic cloning and expression of DNA-Restriktions-/Modifikationssystemen program. So we have for years set the standards in terms of quality and price. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/3/34/Bonn_sponsors_MN.png class=bottom-sponsor width=250px id=sponsor-mn> </td> <td> <h2> Macherey und Nagel </h2> </br> MACHEREY-NAGEL is a family-run concern in the fourth generation. The comprehensive portfolio includes the areas of filtration, rapid tests, water analysis, chromatography and bioanalysis. MACHEREY-NAGEL employs more than 470 highly skilled employees in sales, production as well as research and development, including 10% post-doctoral researchers. They all guarantee an exceptional service. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/3/37/Bonn_Sponsor_roche.jpg class=bottom-sponsor width=250px id=sponsor-roche> </td> <td> <h2> Roche </h2> </br> Roche Headquartered in Basel, Switzerland, is a leading research-focused healthcare company with the pharmaceuticals and diagnostics businesses. As the worlds largest biotech company developing clinically differentiated medicines in oncology, virology, inflammation, metabolism and central nervous system. Roche, a pioneer in diabetes management, is also the world leader in in-vitro diagnostics, tissue-based cancer diagnostics. Medicines and diagnostic tools that enable tangible improvements in the health, quality of life and survival of patients is the strategic goal of personalized medicine from Roche. This concept is based on new molecular insights and molecular diagnostic tests that allow a more precise tuning of therapy and better control of the disease. Therapies are tailored to patient groups that have similarities in their disease. The only way to improve the efficacy of drugs targeted and maintain quality of life. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/d/d2/Bonn_sponsor_geneious.gif class=bottom-sponsor width=250px id=sponsor-geneious> </td> <td> <h2> Geneious </h2> </br> In Good Company</br> First released in 2005, Geneious is one the worlds leading bioinformatics software platforms, used by over 2500 universities and institutes and commercial companies in more than 65 countries. Geneious is used by all 20 of the top 20 Universities globally (Times Higher Education, 2012) and by seven of the 10 largest pharmaceutical companies.</br> Dedicated to excellence</br> Our software has won a number of prestigious awards, including the Computerworld Excellence Awards from Innovative Use of ICT and the United Nations World Summer Awards and Winner in the e-Science Category in 2007, the Recruit IT Innovative Software Product Award at the PriceWaterhouseCoopers Hi-Tech Awards in 2009 and a Global Finalist in the IT and Informatics category at the Bio-IT World Awards in Boston in 2009. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/5/55/Bonn_Sponsor_vwr.jpg class=bottom-sponsor width=250px id=sponsor-vwr> </td> <td> <h2> VWR </h2> </br> VWR is science for the advancement of the worlds most important research through the distribution of a wide range of products and services to a variety of important companies in the pharmaceutical, biotechnology, and healthcare industries as well as government agencies, universities and schools. We offer our customers all the resources they need to be successful, ie an extensive range of the best products in the areas of chemicals, furniture, appliances, instruments, apparel and consumables, from a wide variety of leaders in the field of science manufacturers. With 160 years of experience in this industry, VWR further supports its customers through a combination of strength, vision, innovation and a well-established distribution network that reaches thousands of specialized labs and facilities across the planet. VWR is not just a product supplier - it keeps the most important research in the world in motion. VWRs expertise in the areas of supply chain and logistics services enables customers to fully concentrate on their areas of expertise. Of the management of procurement processes to the integration of supply chains: VWR helps specialized research facilities and laboratories to work with maximum efficiency. VWR has over 8,000 employees in 30 countries with direct offices throughout the world working to streamline the way, as researchers from North America, Europe and the Asia-Pacific region supply and maintain their labs. In addition, VWR further supports its customers by providing onsite services, storeroom management, product procurement, supply chain systems integration and technical services. We are expanding our global presence and adhere to the principle that customers benefit from the availability and expertise of our local sales teams.</br> In todays economy VWR helps its customers to focus on increasing productivity and reducing costs and optimizing procurement processes. </br> Headquartered in Radnor, PA (USA), earned VWR International, LLC, in 2012 global sales of more than 4.1 billion U.S. dollars. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/0/0b/Bonn_Sponsors_Roth.png class=bottom-sponsor width=170px id=sponsor-roth> </td> <td> <h2> ROTH – A COMPANY WITH TRADITION </h2> </br> 1879 </br> Carl ROTH founded in Karlsruhe, a "material, Colonial and dye business and Droguerie".</br>1899</br> The first sales and mail order catalog is published.</br> 1956</br> Publication of the first issue of the publication a "Rarea " Natural Products</br> 1990</br> First ROTH general catalog. Complete with the areas of laboratory, life sciences and chemicals in one.</br> 2005</br> Completion of a modern establishment for the production and storage of laboratory chemicals and reagents in Karlsruhe Rhine port area. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/7/79/Bonn_starlab.jpg class=bottom-sponsor width=250px id=sponsor-starlab> </td> <td> <h2> Starlab </h2> </br> STARLAB is a company specializing in liquid handling technology group. With subsidiaries in Germany, France, Britain and Italy is available in the direct sales an extensive range of products available. Plus, you get our products to many countries around the world via our international trading partners. Our success is based on many years of experience in manufacturing and marketing of liquid handling disposable products - with TipOne we have established ourselves as a leading supplier of pipette tips systems worldwide. This quality, price and service come first. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/b/bf/Bonn_caesar.gif class=bottom-sponsor width=200px id=sponsor-casar> </td> <td> <h2> Caesar </h2> </br> The center of advanced european studies and research (caesar) is an institute of the Max Planck Society, which is located at the boundaries between neuroscience, cell biology and biophysics. The focus of the research is the particular cellular and neural signal processing. </br> Caesar works with modern photonic, molecular biological, chemical and micro-technological methods. The focus of kinetic, spectroscopic and microscopic methods are research and control of cellular activity. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/6/6e/Bonn_Eurofins.png class=bottom-sponsor width=250px id=sponsor-eurofins> </td> <td> <h2> Eurofins MWG operon </h2> </br> Discover our fascination about the world of the four bases.</br> We are fascinated about the power of DNA and how it is incorporated in everything we do, work and live. Being passionate about our strong customer orientation, our service and our quality standards, we continuously challenge ourselves to stay ahead and remain one of the leading genomics service providers worldwide. Eurofins MWG Operon is globally known for its innovative and customised technologies in the life science industries and academic research institutions. With the combined power of an international network of Eurofins companies in the field of genomic services, forensics, agroscience, pharmaceutical, environmental, food and feed testing, we have established an outstanding team of experts and broad range of technologies. This unique constellation underlines our approach to offer best practise solutions and versatile concepts for our clients - academic institutions and large interdisciplinary operating companies of the world. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/1/18/Bonn_ThermoFisher.jpg class=bottom-sponsor width=250px id=sponsor-thermofisher> </td> <td> <h2> Thermo Fisher </h2> </br> Thermo Fisher Scientific Inc. (NYSE: TMO) is the world leader in serving science. Our mission is to enable our customers to make the world healthier, cleaner and safer. With revenues of $13 billion, we have approximately 39,000 employees and serve customers within pharmaceutical and biotech companies, hospitals and clinical diagnostic labs, universities, research institutions and government agencies, as well as in environmental and process control industries. We create value for our key stakeholders through three premier brands, Thermo Scientific, Fisher Scientific and Unity<sup> TM </sup> Lab Services, which offer a unique combination of innovative technologies, convenient purchasing options and a single solution for laboratory operations management. Our products and services help our customers solve complex analytical challenges, improve patient diagnostics and increase laboratory productivity. </td></tr></table> </div> </div> </div> </div> </div> ";

content.type="Team";

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content.childs=[];

content.titleShort = "Bonn";

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content.titleLong = "Bonn";

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content.titleLong = "Our city Bonn";

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content.summary= "~~Description of~~ Bonn";

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content.summary= "Bonn is a beautiful city in North Rhine-Westphalia, Germany.";

-

content.text= "~~Founded~~ by the Romans in the year 12 before Christ, birthplace of Beethoven~~, once~~ Capitol of Germany~~- now:~~ Bonn ~~is a vivid place to life~~ and ~~study~~. ~~Next to Cologne, Bonn~~ is ~~also~~ set on the river Rhine. Everybody gets smitten with ~~it´s~~ charm~~- such a~~ unique mixture of tradition and modern lifestyle.</br>Stadt. City. Ville. Bonn.";

+

content.text= "Bonn has a long and eventful history. The city was founded by the Romans in the year 12 before Christ and continued to grow ever since. Bonn also was the birthplace of the famous musician and composer Beethoven and even was the Capitol of West Germany for 41 years. Today Bonn has around 300 000 habitants and has become the cultural of the region. It is set on the river Rhine very close to Cologne. Everybody gets smitten with its charm and unique mixture of tradition and modern lifestyle. The city's historic center and the setting close to the river Rhine turn it into a wonderful place to live and study.<p style='text-align:center;'>the city's slogan:</br><i>"Stadt. City. Ville. Bonn"</i></p></br><p><a href=https://static.igem.org/mediawiki/2013/0/0b/Bonn_by_night.JPG><div class='content-image'align='center'><img src='https://static.igem.org/mediawiki/2013/0/0b/Bonn_by_night.JPG' width='600'></a>Bonn by night <sup>©Jan Hansen</sup></div></p><p><div class='content-image'align='center'><a href=https://static.igem.org/mediawiki/2013/a/a6/Bonn_poppelsdorfer_schloss.JPG><img src='https://static.igem.org/mediawiki/2013/a/a6/Bonn_poppelsdorfer_schloss.JPG' width='600'></a>The university <sup>©Jan Hansen</sup></div><p><a href=https://static.igem.org/mediawiki/2013/0/0b/Bonn_by_night.JPG><div class='content-image'align='center'><img src='https://static.igem.org/mediawiki/2013/7/75/Bonn_beethoven.JPG' width='600'></a>The famous Beethoven statue in the city center <sup><a href=http://www.bone-diagenesis.org/moziloCMS/gallery.php?gal=Bonn-nbsp~Impressions>[source]</a></sup></div></p><h2>References</h2><p><a name='1'>1.</a> <a href='http://www.bonn.de'>http://www.bonn.de</a></p><p><a name='2'>2.</a> <a href='http://de.wikipedia.org/wiki/Bonn'>http://de.wikipedia.org/wiki/Bonn</a></p>";

content.type="Team";

break;

Line 990:

Line 905:

break; */

-

+

/*

case 112:

content.i = 112;

-

content.parents = [100~~, 109~~];

+

content.parents = [100];

-

content.childs = [];

+

content.childs = [109];

content.titleShort = "Comic";

content.titleLong = "Comic – The adventures of Obi Wan E.Coli";

Line 1,001:

Line 916:

content.type = "Human Practice";

break;

+

*/

+

case 200:

+

content.i = 200;

+

content.parents=[37];

+

content.childs=[];

+

content.titleShort = "Safety";

+

content.titleLong = "Safety";

+

content.summary= "Since safety is an important concern for any iGEM team, we gave deep thought to it. In this article our reflections about safety measures are presented.";

+

content.text= " Safety is an important issue for all of the IGEM teams. Taking care of the health and safety of our teammates is of the utmost importance for us. In our opinion, only a safe lab is a good environment for research and work. On our safety page we would like to give you a brief overview over the precautions we take to ensure a safe work environment in the lab. </br></br> <h2> A </h2> Do the biological materials used in your lab work pose any of the following risks: </br> </br> 1. Risks to the safety and health of team members or others working in the lab? </br> In our lab, we work with E. coli (strain: DH5alpha;XL1Blue;w3110 sspB-; w3110 sspB- LacI-DAS+4;Top10; DH10 Beta). All these E.Coli strains are classified as risk group 1, as they can cause irritation of the skin, the eyes, and the respiratory tract; additionally, they can affect the kidneys. Although our E.coli strains are nonpathogenic, we wear protective clothes and examination gloves to ensure safety. </br> We also use ethidum bromide in our lab, which is toxic and can cause cancer. To minimize the risk of accidental contact with this dangerous substance, we have fume hoods especially designated for working with ethidium bromide. Furthermore, the use of nitrile gloves as well as lab coats is mandatory when we are working with potentially harmful substances and any waste contaminated with ethidium bromide is kept separate from other waste. </br> In addition, every team member had to take a course about biosafety and regulations in Germany (as required in §8-12 of the GenTSV, the German act of genetic engineering). </br></br> 2. Risks to the safety and health of the general public, if released by design or by accident? </br> There is little risk to the general public, since our bacteria strains are nonpathogenic and in general do not cause harm to humans. And yet, the contact of an immunocompromised person with our bacteria strains could lead to health issues. </br></br> 3. Risk to the environment, if released by design or by accident? </br> Since our bacteria and genetic parts are non pathogenic and the light induced protein degradation is not a threat to the environment, there is minimal risk to the environment if our (modified) bacteria were released . In addition, our lab is equipped with a special air conditioning system which maintains a slight low pressure environment in the lab and thus prevents the air from inside of the lab from getting out when the doors are opened. The risk of (genetically altered) bacteria being released by accident is therefore reduced. </br> </br> 4.Risks to security through malicious misuse by individuals, groups, or countries? </br> The risk of malicious misuse can never be completely eliminated. Although there are cameras around the campus as well as other security installations, an element of risk remains. If our project was stolen and our bacteria were released, the risks for the public would be minimal, since our bacteria strains are nonpathogenic and our project mainly deals with the light-inducible degradation of proteins, which is hardly dangerous. </br></br> 5. If your project moved from a small-scale lab study to become widely used as a commercial/industrial product, what new risks might arise? </br> Since our light-inducible degradation system is mostly thought to be applicable in restricted areas (labs, bioreactors) the risks are low. Of course, any industrial application of our system should mot be designed without additional fail-safes to ensure multiple layers of safety. </br> One possible “risk” of our project is inherent to any other biological system; namely, that mutations in our device's gene sequence lead to unforeseen consequences. We assume that it is most likely that a mutation in our device will lead to the device's failure, i.e. leading to an accumulation of the protein that is supposed to be degraded. Whether this poses merely efficiency problems or more serious biosafety risks depends on the specific application and should be considered by persons responsible for implementing that instance of our technology. </br></br> 6. Does your project include any design features to address safety risks? (For example: kill switches, auxotrophic chassis, etc.) Note that including such features is not mandatory to participate in iGEM, but many groups choose to include them. </br> We developed a kill switch that is induced by blue light. This switch is capable of killing bacteria that are accidentally removed from a restricted area - for example a red-light hood – and which are then exposed to daylight in a short period of time. </br></br> 7. What safety training have you received (or plan to receive in the future)? Provide a brief description, and a link to your institution’s safety training requirements, if available. </br> Every team member had to take a course about safe work in the lab. We also have advisors who teach us how to work properly and safely and inform us about safety measures, such as wearing protective clothing. We also plan to implement a disinfection checkpoint that our team members will have to pass when leaving our entering the lab to decrease the chance of contamination. </br></br> 8. Under what biosafety provisions will / do you work? </br> Please provide a link to your institution biosafety guidelines. </br> The institution's biosafety guidelines are the national safety guidelines. <sup><a href=#2001>200.1</a></sup> </br> </br></br> <h2>B</h2> Does your institution have an Institutional Biosafety Committee, or an equivalent group? If yes, have you discussed your project with them? Describe any concerns they raised with your project, and any changes you made to your project plan based on their review. </br> Our university does not have a committee for biosafety. We are using the guidelines from the central biosafety committee from our federal government. We also work together with our supervisor, discussing our project and to address any safety issues that might arise. </br></br></br> <h2>C</h2> Does your country have national biosafety regulations or guidelines? If so, please provide a link to these regulations or guidelines if possible. <sup><a href=#2001>200.1</a></sup> <sup><a href=#2002>200.2</a></sup> </br></br></br> <h2>D</h2> According to the WHO Biosafety Manual, what is the BioSafety Level rating of your lab? </br> Our Biosafety Level is 1. <h2><b>References</b></h2></br> <a name=2001>200.1</a> <a href= http://www.gesetze-im-internet.de/gentsv/index.html#BJNR023400990BJNE001503320> http://www.gesetze-im-internet.de/gentsv/index.html#BJNR023400990BJNE001503320 </a> </br> <a name=2002>200.2</a> <a href= http://www.gesetze-im-internet.de/gentsv/anhang_iii_34.html> http://www.gesetze-im-internet.de/gentsv/anhang_iii_34.html </a>";

+

content.type="Project";

+

break;

+

}

+

}

</script>

</html>

@@ Line 34: / Line 34: @@
 content.i = 2;
 content.parents=1;
-content.childs=[3,11,9,10,7];
+content.childs=[4,5,9,11,7,10];
 content.titleShort = "Protein regulation";
 content.titleLong = "Protein regulation mechanisms";
@@ Line 59: / Line 59: @@
 content.i = 4;
-content.parents=[3]
+content.parents=[2]
 content.childs=[];
 content.titleShort = "Protein cleavage";
@@ Line 71: / Line 71: @@
 content.i = 5;
-content.parents=[3];
+content.parents=[2];
 content.childs=[];
 content.titleShort = "Gene knock-out and knock-in";
@@ Line 129: / Line 129: @@
 content.titleShort = "Direct inhibition and activation";
 content.titleLong = "Direct inhibition and activation";
-content.summary= "Direct binding of small molecules an activate or inactivate protein function";
+content.summary= "Direct binding of small molecules in order to activate or inactivate protein function";
-content.text= "Moreover proteins can be regulated directly in many different ways. Binding of other Proteins or small molecules for example can activate or deactivate the catalytic function of proteins. For example small molecules can have great impact on protein function. They can often influence protein functions by binding to an allosteric center. Small molecules can be activating, as well as repressing, depending on the type of the molecule<sup><a href=#11.1>11.1</a></sup>.</br></br><div class='content-image'> <img src=https://static.igem.org/mediawiki/2013/1/1c/BonnSmallMolecules.jpg></br>Different mechanisms for small-molecule activation of enzymes<sup><a href=#11.2>11.2</a></sup> </div></br></br>The following picture shows the example of the allosteric activation of a glucokinase</br> <div class='content-image'> <img src=https://static.igem.org/mediawiki/2013/f/f4/BonnSmallMolecules2.jpg></br>&quot;(a) GK bound to the GKA, compound A, and glucose (blue, Protein Data Bank (PDB) ID 1V4S). Compound A binds at a site distal from the active site, which is highlighted by the presence of the substrate, glucose. (b) Structural overlay of GK in the presence of compound A and glucose with an unliganded, inactive GK (pink, PDB ID 1V4T). In the unbound GK, the GKA binding site is occluded. A large shift in the small subunit of GK occurs from the unbound to bound structures (black arrow). Glucose promotes the active conformation, which is hindered from shifting back to the inactive conformation in the presence of compound A. (c) 7 mutations (out of 13)9 identified in GK (pink) that are associated with disease map to the GKA binding site. These mutations highlight an important regulatory site within GK, and could similarly stabilize a closed, active conformation.&quot;<sup><a href=#11.2>[11.2]</a></sup></div></br></br><p><a name=11.1>11.1</a><a href='http://www.nature.com/nrd/journal/v3/n4/full/nrd1343.html'>Small-molecule inhibitors of proteinprotein interactions: progressing towards the dream; Michelle R. Arkin & James A. Wells</a>/p></br><p><a name=11.2>11.2</a><a href='http://www.nature.com/nchembio/journal/v6/n3/full/nchembio.318.html'>Turning enzymes ON with small molecules, Julie A Zorn & James A Wells</a></p>";
+content.text= "Direct binding of small molecules an activate or inactivate protein function";
+content.text= "Moreover proteins can be regulated directly in many different ways. Binding of other Proteins or small molecules for example can activate or deactivate the catalytic function of proteins. For example small molecules can have great impact on protein function. They can often influence protein functions by binding to an allosteric center. Small molecules can be activating, as well as repressing, depending on the type of the molecule<sup><a href=#11.1>11.1</a></sup>.</br></br><div class='content-image'> <img src='https://static.igem.org/mediawiki/2013/1/1c/BonnSmallMolecules.jpg' width='550'> </br>Different mechanisms for small-molecule activation of enzymes<sup><a href=#11.2>11.2</a></sup> </div></br></br>The following picture shows the example of the allosteric activation of a glucokinase</br> <div class='content-image'> <img src=https://static.igem.org/mediawiki/2013/f/f4/BonnSmallMolecules2.jpg width='600'></br>&quot;(a) GK bound to the GKA, compound A, and glucose (blue, Protein Data Bank (PDB) ID 1V4S). Compound A binds at a site distal from the active site, which is highlighted by the presence of the substrate, glucose. (b) Structural overlay of GK in the presence of compound A and glucose with an unliganded, inactive GK (pink, PDB ID 1V4T). In the unbound GK, the GKA binding site is occluded. A large shift in the small subunit of GK occurs from the unbound to bound structures (black arrow). Glucose promotes the active conformation, which is hindered from shifting back to the inactive conformation in the presence of compound A. (c) 7 mutations (out of 13)9 identified in GK (pink) that are associated with disease map to the GKA binding site. These mutations highlight an important regulatory site within GK, and could similarly stabilize a closed, active conformation.&quot;<sup><a href=#11.2>[11.2]</a></sup></div></br></br><p><a name=11.1>11.1</a><a href='http://www.nature.com/nrd/journal/v3/n4/full/nrd1343.html'>Small-molecule inhibitors of proteinprotein interactions: progressing towards the dream; Michelle R. Arkin & James A. Wells</a>/p></br><p><a name=11.2>11.2</a><a href='http://www.nature.com/nchembio/journal/v6/n3/full/nchembio.318.html'>Turning enzymes ON with small molecules, Julie A Zorn & James A Wells</a></p>";
 content.type="Background";
 break;
@@ Line 166: / Line 167: @@
 content.titleLong = "Ec. SspB adaptor";
 content.summary= "The sspB protein is an adaptor responsible for delivering ssrA-tagged substrates to the ClpXP protease in order to enhance their degradation";
-content.text= "<b>Introduction </b> </br> The sspB protein is an adaptor responsible for delivering ssrA-tagged substrates to the ClpXP protease in order to enhance their degradation.Thus, bacterias like E.coli or C.crescentus regulate the concentration of marked proteins and also are in control of their quality. Even though degeneration of tagged substrates is possible without sspB, sspB delivering is a common process, because it improves the affinity between ssrA and ClpXP<sup><a href='#14.1'>14.1</a></sup>. </br> In our project, we used a sspB split variant instead of the normal sspB in order to control cleaving rate. The two parts of this version only stay divided until we ray them with light of a special wavelength. After that, the fractions form an unit and can function normal from now on as described below. </br> </br><b>Structure </b> </br>The sspB adaptor is a homomeric dimer (Fig. 1), which means that itconsists of two identical domains. Together the domains form a pore with the ssrA binding sites inside. Each domain also owns a C-terminal tail ending in a ClpX binding module, called XP. The amino-acid sequence of XP is highly conserved so that mutations in it mostly cause extremely decrease in activity, whereas the linker sequence differs from species to species <sup><a href='#14.2'>14.2</a></sup><sup><a href='#14.3'>14.3</a></sup>. <div align='left'><img src='https://static.igem.org/mediawiki/2013/5/5c/BonnsspB_Fig2.jpg' height='202' width='403'>Fig. 1: Ribbon diagramm of the sspB dimer with a bound srrA-tagged protein, from 'Versatile modes of peptide recognition by the AAA+ adaptor protein SspB, Levchenko et al, 2005, nature structural and molecular biology, PMID: 15880122</div> </br> </br></br></br><b>Function</b></br> SspB enhances degradation of ssrA-tagged proteins by lowering the K<sub>M</sub>. Thus, with a given substrate concentration sspB-mediated cleaving runs faster than without sspB (Fig. 2) </br> <div align='left'><img src='https://static.igem.org/mediawiki/2013/d/d8/BonnSspB_Fig1.jpg' height='232' width='371'>Fig. 2: Diagramm, shows the degradation rate of GFP-ssrA without sspB, with sspB and with two mutations, the given substrate concentration is 0.3 &my;M, from 'Flexible Linkers Leash the Substrate Binding Domain of SspB to a Peptide Module that Stabilizes Delivery Complexes with the AAA ClpXP Protease, Wah et al, Molecular Cell, 2003, PMID: 14536075' </div></br> Therefor, the sspB dimer contains a pore and while 'AADENY' is linked with the inside, the 'LAA'-domain (respectively 'DAS') faces outwards, free to bind ClpX (Fig. 3).</br> The affinity of this binding amounts around 20 &my;M, which suggests a relative strong  connection. The two extremely flexible ClpX binding tails with XP at the C-terminal end  dock on ClpX. So the 'LAA'- domain lies closely to ClpX's axial pore and can be bound to it. <div align='left'><img src='https://static.igem.org/mediawiki/2013/7/7c/BonnSsra_fig3.jpg' height='151' width='116'>Fig. 3: Model of sspB with a bound ssrA-tagged substrate, from 'Engineering controllable protein degradation, McGinness et al, Molecular cell, 2006, PMID:16762842' </div></br> To sum up, there are three bonds connecting the ssrA-sspB-ClpX-complex and making it relative stable: ssrA with sspB, sspB with ClpX and ssrA with ClpX. Hence follows a lower K<sub>M</sub> than the direct binding process has<sup><a href='#14.4'>14.4</a></sup><sup><a href='#14.5'>14.5</a></sup>. <h2> References </h2> <a id='14.1'>14.1</a>Bivalent Tethering of SspB to ClpXP Is Required for Efficient Substrate Delivery: A Protein-Design Study, Bolon et al, 2004, Molecular Cell, PMID: 14967151 </br><a id='14.2'>14.2</a>see above </br> <a id='14.3'>14.3</a> Flexible Linkers Leash the Substrate Binding Domain of SspB to a Peptide Module that Stabilizes Delivery Complexes with the AAA ClpXP Protease, Wah et al, Molecular Cell, 2003, PMID: 14536075 </br> <a id='14.4'>14.4</a> see above</br> <a id='14.6'>[14.5]</a> Altered Tethering of the SspB Adaptor to the ClpXP Protease Causes Changes in Substrate Delivery, McGinnes KE et al, The journal of Biological Chemistry, 2007, PMID: 17317664 </br> <a id='14.4'>14.4</a>  ClpXP, an ATP-powered unfolding and protein-degradation machine, Baker et al, Biochim Biophys Acta, 2012, PMCID: PMC3209554</br>";
+content.text= "<b>Introduction </b> </br> The sspB protein is an adaptor responsible for delivering ssrA-tagged substrates to the ClpXP protease in order to enhance their degradation.Thus, bacterias like E.coli or C. crescentus regulate the concentration of marked proteins and also are in control of their quality. Even though degeneration of tagged substrates is possible without sspB, sspB delivering is a common process, because it improves the affinity between ssrA and ClpXP<sup><a href='#14.1'>14.1</a></sup>. </br> In our project, we used a sspB split variant instead of the normal sspB in order to control cleaving rate. The two parts of this version only stay divided until we ray them with light of a special wavelength. After that, the fractions form an unit and can function normal from now on as described below. </br> </br><b>Structure </b> </br>The sspB adaptor is a homomeric dimer (Fig. 1), which means that itconsists of two identical domains. Together the domains form a pore with the ssrA binding sites inside. Each domain also owns a C-terminal tail ending in a ClpX binding module, called XP. The amino-acid sequence of XP is highly conserved so that mutations in it mostly cause extremely decrease in activity, whereas the linker sequence differs from species to species <sup><a href='#14.2'>14.2</a></sup><sup><a href='#14.3'>14.3</a></sup>. <div align='left'><img src='https://static.igem.org/mediawiki/2013/5/5c/BonnsspB_Fig2.jpg' height='202' width='403'>Fig. 1: Ribbon diagramm of the sspB dimer with a bound srrA-tagged protein, from 'Versatile modes of peptide recognition by the AAA+ adaptor protein SspB, Levchenko et al, 2005, nature structural and molecular biology, PMID: 15880122</div> </br> </br></br></br><b>Function</b></br> SspB enhances degradation of ssrA-tagged proteins by lowering the K<sub>M</sub>. Thus, with a given substrate concentration sspB-mediated cleaving runs faster than without sspB (Fig. 2) </br> <div align='left'><img src='https://static.igem.org/mediawiki/2013/d/d8/BonnSspB_Fig1.jpg' height='232' width='371'>Fig. 2: Diagramm, shows the degradation rate of GFP-ssrA without sspB, with sspB and with two mutations, the given substrate concentration is 0.3 &my;M, from 'Flexible Linkers Leash the Substrate Binding Domain of SspB to a Peptide Module that Stabilizes Delivery Complexes with the AAA ClpXP Protease, Wah et al, Molecular Cell, 2003, PMID: 14536075' </div></br> Therefor, the sspB dimer contains a pore and while 'AADENY' is linked with the inside, the 'LAA'-domain (respectively 'DAS') faces outwards, free to bind ClpX (Fig. 3).</br> The affinity of this binding amounts around 20 &my;M, which suggests a relative strong  connection. The two extremely flexible ClpX binding tails with XP at the C-terminal end  dock on ClpX. So the 'LAA'- domain lies closely to ClpX's axial pore and can be bound to it. <div align='left'><img src='https://static.igem.org/mediawiki/2013/7/7c/BonnSsra_fig3.jpg' height='151' width='116'>Fig. 3: Model of sspB with a bound ssrA-tagged substrate, from 'Engineering controllable protein degradation, McGinness et al, Molecular cell, 2006, PMID:16762842' </div></br> To sum up, there are three bonds connecting the ssrA-sspB-ClpX-complex and making it relative stable: ssrA with sspB, sspB with ClpX and ssrA with ClpX. Hence follows a lower K<sub>M</sub> than the direct binding process has<sup><a href='#14.4'>14.4</a></sup><sup><a href='#14.5'>14.5</a></sup>. <h2> References </h2> <a id='14.1'>14.1</a>Bivalent Tethering of SspB to ClpXP Is Required for Efficient Substrate Delivery: A Protein-Design Study, Bolon et al, 2004, Molecular Cell, PMID: 14967151 </br><a id='14.2'>14.2</a>see above </br> <a id='14.3'>14.3</a> Flexible Linkers Leash the Substrate Binding Domain of SspB to a Peptide Module that Stabilizes Delivery Complexes with the AAA ClpXP Protease, Wah et al, Molecular Cell, 2003, PMID: 14536075 </br> <a id='14.4'>14.4</a> see above</br> <a id='14.6'>[14.5]</a> Altered Tethering of the SspB Adaptor to the ClpXP Protease Causes Changes in Substrate Delivery, McGinnes KE et al, The journal of Biological Chemistry, 2007, PMID: 17317664 </br> <a id='14.4'>14.4</a>  ClpXP, an ATP-powered unfolding and protein-degradation machine, Baker et al, Biochim Biophys Acta, 2012, PMCID: PMC3209554</br>";
 content.type="Background";
 break;
@@ Line 211: / Line 212: @@
 content.i = 18;
 content.parents=[17];
-content.childs=[19,21,20];
+content.childs=[36,21,20];
 content.titleShort = "Light";
 content.titleLong = "Light as a means of induction";
@@ Line 219: / Line 220: @@
 break;
+/*
 case 19:
@@ Line 231: / Line 232: @@
 content.type="Background";
 break;
+*/
+/*
 case 20:
@@ Line 244: / Line 246: @@
 content.type="Background";
 break;
+*/
 case 21:
@@ Line 302: / Line 304: @@
 content.i = 36;
-content.parents=[19];
+content.parents=[18];
 content.childs=[37];
 content.titleShort = "LOV-ipaA & VinD1";
@@ Line 308: / Line 310: @@
 content.summary= "The LOV-ipaA -vinculin system is a combined system for light inducible heterodimerisation. This powerful tool, which allows photocontroled complex formation was establish by Lungu et al. in 2012.";
 content.text= "The LOV-ipaA -vinculin system is a combined system for light inducible heterodimerisation. It consists out of a LOV domain, which undergoes conformational changes upon irradiation with blue light, and the ipaA-vinculin hybridization system. This two building blocks have be combined and described by Lungu et al. in 2012.</br></br> Lungu et al. (2008) where able to measure a 49-fold difference in target binding upon irradiation as compared to the dark state. However, they further modified the system by mutations of the LOV-ipaA construct and successfully weakend the baseline affinity for vinculin (initial design: 3.5 nM to 69 nM; mutant: 2.4 nM to >40µM affinity for vinculin) to reduce the dark state activity. </br></br> Lov-ipaA-VinD1 is a powerful tool which allows photocontroled complex formation. To establish this system Lungu et al. (2012)<sup><a href=#361>36.1</a></sup> fused the Ja helix of the LOV Domain to ipaA.</br>To be more precise they used the LOV2 domain from Avena sativa photopropin 1 (AsLOV2), which – as previously shown – can be used to photomodulate the affinity of peptides for their binding partners (see Figure 1). </br><div class=contant-image><img src=https://static.igem.org/mediawiki/2013/0/02/Bonn_MS_Figure1_LOV.jpg></br>Figure 1: General design of AsLOV2 fusion proteins (Lungu et al. 2012)<sup><a href=#361>36.1</a></sup></div> </br>In other studies had been shown that the LOV domain can be fused to entire protein domains, allowing photomodulation of the protein binding. However, they stated that it might be of high importance to bring the LOV domain closer to ipaA, in order to allow photomodulation in this case, because ipaA is only a peptide and thus more flexible than folded domains.</br></br>Therefore, Lungu et al. (2012)<sup><a href=361>36.1</a></sup> identified similar amino acid sequences in the ipaA peptide and the Ja helix of the LOV Domain and used this combined with molecular modeling to create photomodulateable AsLOV2-ipaA (see Figure 2). </br><div class=contant-image><img src=https://static.igem.org/mediawiki/2013/5/54/Bonn_MS_Figure2_LOV-ipaA.jpg></br>Figure 2: Light-inducible LOV-ipaA construct (Lungu et al. 2012)<sup><a href=361>36.1</a></sup></div></br>They were able to proof the functionality of the AsLOV2-ipaA system by heterodimerization in yeast (yeast two-hybrid system or Y2H) The yeast two-hybrid system can be used to monitor protein–protein interactions between two proteins. The system is based on a transcription factor, which is split into two separate fragments, called the binding domain (BD) and activating domain (AD). Each domain is fused to one protein and thus only if the proteins interact with each other BD and AD are close enough to initiate the transcription of a reporter gene.</br></br>The basic principle of the LOV-ipaA & VinD1 system works as follows. In the dark state the fusion product LOV-ipaA is not able to form a complex with vinculin, because LOV blocks ipaA sterically. However, activation of the LOV domain with blue light induces conformational changes in the fused molecule, which results in a movement of the Ja helix with the ipaA away from LOV. Thereby, ipaA becomes accessible for VinD1 and a Complex is formed.</br></br>The activation is reversible and the entire system can be genetically encoded. This two facts are the main advantages of this system in contrast to other typically used systems, which like for the chemical system for example, are based on in vivo covalently modified peptides, that can be activated by light induced cleavage. Moreover, the protein used are relatively small and thus should interfere as little as possible with the prokaryotic metabolism, the activity change form dark to light state is high, the system is completely genetically encoded and reversible. But, the most important property of this system is that it allows the light-controlled heterodimerisation of the two split variants of sspB, which is necessary for our system.</br></br><h2>References</h2></br><a name=361>36.1</a> <a href=http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3334866/>Lungu et al. (2012) Designing photoswitchable peptides using the AsLOV2 domain</a>";
-content.type="Project";
+content.type="Background";
 break;
@@ Line 316: / Line 318: @@
 content.i = 37;
 content.parents=[36,21,12];
-content.childs=[38,45,53,69,52,57,65,100];
+content.childs=[38,45,53,69,52,57,65,100,200];
 content.titleShort = "Project summary";
 content.titleLong = "Our project in a nutshell";
-content.summary= "We engineer a photoswitchable protein degradation system relying on the ClpXP protease system and a LOV domain from avena sativa";
+content.summary= "We engineer a photoswitachble protein degradation system relying on the ClpXP protease system an a LOV domain from avena sativa";
-content.text= "";
+content.text= "A reliable, yet easily adaptable mechanism for controlling protein activity is key to most areas of life and medical science research. Still, the most common approaches suffer from various flaws. Knocking genes out using homologous recombination, knocking a gene down with RNA interference or modulating the behaviour of a protein with a chemical stimulus - just to name a few prominent methods - is either restricted to non-lethal genes, does not yield a big difference in activity, or is absolutely inaccurate and thus prone to secondary effects.</br></br> Would it not be great if one could turn off any protein, at any time, with little to no side effects? That is where iGEM Bonn 2013 and their project comes in. We aim to overcome the aforementioned difficulties by engineering a novel tool based on blue-light-inducible degradation of targeted proteins.</br></br>Our system relies on two key components: A tiny (just 15 amino acids!) tag that is fused to the C-Terminus of a protein of your choosing, and a light sensing LOV (Light, Oxygen and Voltage) domain from Avena sativa.</br></br> The advantages of our approach are obvious: Not only does the usage of light allow for a superior tempero-spatial control, but it is also much less prone to unwanted side effects than any chemical stimulus.</br> Furthermore, as we rely on a direct degradation of the targeted protein, we expect an onset of the desired effect which is much faster and at least as high as in common approaches.</br> Finally, as our system requires only a minor modification of your target protein we expect its function to not be impaired, and the tag to go unnoticed in functional observations.";
-content.type="Project";
+content.type="Projekt";
 break;
@@ Line 355: / Line 357: @@
 content.childs=[41,74,43];
 content.titleShort = "C. crescentus";
-content.titleLong = "General information about C. crescentus and the ClpXP protein degradation system";
+content.titleLong = "Ortholog ClpXP system in C. crescentus";
-content.summary= "Here you can find a brief introduction to Caulobacter crescentus and its ClpXP protease system.";
+content.summary= "Using the ortholog ClpXP protein degradation system of C. crescentus in E. coli enables us to induce protein degradation in native E. coli without interfering with their natural system.";
 content.text= "Caulobacter crescentus is a Gram-negative &alpha;-protobacterium often found in fresh water lakes or in the sea. Its cell division cycle is a favoured object of study due to its remarkable asymmetry. <sup><a href=401>40.1</a></sup></br>ClpXP is a protease that degrades proteins tagged with the ssrA peptide. sspB&alpha; is an adaptor protein that can accelerate protein degradation by tethering ssrA-tagged proteins towards the ClpX subunit of the ClpXP protease.</br>For further information, please browse the related articles. </br></br><h2>References</h2></br><a href=401>40.1</a><a href=’ http://www.amazon.com/Brock-Biology-Microorganisms-13th-Edition/dp/032164963X’> Brock Microbiology of Microorganisms, Madigan et al., Pearson, German edition Vol. 13, 2012</a>";
 content.type="Project";
@@ Line 365: / Line 367: @@
 content.parents=[40];
 content.childs=[];
-content.titleShort = "C. crescentus ClpXP";
+content.titleShort = "<sup>Cc</sup>ClpXP";
-content.titleLong = "C. crescentus ClpXP";
+content.titleLong = "C. crescentus: ClpXP";
 content.summary= "This article is about the ClpXP protease system in C. crescentus and regulation of proteolysis via ClpXP. It also provides information about similarities and differences between C. crescentus and E. coli orthologs of related proteins.";
 content.text= "<div class='content-image'><img src='https://static.igem.org/mediawiki/2013/d/de/Bonn-41-CC-ClpXP.png'>ClpXP mediated proteolysis <sup><a href=#413>41.3</a></sup></div>ClpXP is an AAA+ protease which is of particular importance for proper cell-cycle progression. It consists of a hexameric ClpX subunit, which recognizes and unfolds tagged proteins, while ATP is hydrolyzed, and a ClpP subunit, that contains the actual peptidase domain. <sup><a href=#411>41.1</a>, <a href=#412>41.2</a></sup> Its structure is highly conserved, such that E. coli and C. crescentus orthologs are very similar. </br>Specific activity of the ClpXP protease is mediated by sspB and ssrA. ssrA is a short peptide consisting of fourteen amino acids in C. crescentus. Proteins which need to be degraded, e.g. for regulatory purpose or due to errors in their structure, are tagged with the ssrA peptide that can be recognized by the ClpX subunit. sspB is a dimeric adaptor protein in C. crescentus which tethers ssrA-tagged proteins to the ClpXP protease and, in this way, accelerates protein degradation. </br>Whilst sspB protein structure is well- conserved among many microorganisms, the structure of <sup>CC</sup>sspB&alpha; (i.e., the C. crescentus ortholog of sspB) shows significant differences compared to other orthologs. For example, <sup>CC</sup>sspB&alpha; and <sup>EC</sup>sspB (i.e., the E. coli ortholog) only show up with sequence identities of 16%, while CCsspB is still able to specifically bind to <sup>EC</sup>ClpXP<sup><a href=#412>41.2</a></sup>. SsrA tags can also be very different among microorganismic species. </br>This makes it possible to establish a protein degradation system in E. coli involving <sup>CC</sup>sspB&alpha; and <sup>EC</sup>ClpXP, as long as the ssrA tag can be recognized by both <sup>CC</sup>sspB&alpha; and <sup>EC</sup>ClpXP. </br></br><h2>References</h2></br><a name=411>41.1</a> <a href=’ http://www.ncbi.nlm.nih.gov/pubmed/17937918’>Structure and substrate specificity of an SspB ortholog: design implications for AAA+ adaptors, Chien et al., Cell Press, 2007, PMID: 17937918</a></br><a name=412>41.2</a> <a href=’http://www.ncbi.nlm.nih.gov/pubmed/20014030’>Versatile modes of peptide recognition by the ClpX N domain mediate alternative adaptor-binding specificities in different bacterial species, Chowdhury et al., Protein Science, 2010, PMID: 20014030</a></br><a name=413>41.3</a> <a href='http://www.biochem.umass.edu/faculty/peter-chien'>Peter Chien, Department of Biochemistry and Molecular Biology (official website), http://www.biochem.umass.edu/faculty/peter-chien, picture URL: cchttp://www.biochem.umass.edu/sites/biochem/files/resize/ClpX-300x268.jpg</a>";
@@ Line 376: / Line 378: @@
 content.parents=[74];
 content.childs=[];
-content.titleShort = "SspB&alpha;";
+content.titleShort = "<sup>Cc</sup>sspB&alpha;";
-content.titleLong = "C. crescentus sspB&alpha;";
+content.titleLong = "C. crescentus: sspB&alpha;";
 content.summary= "This article deals with the structure of sspB&alpha; and conformational details of its binding to ssrA and ClpXP during tethering.";
 content.text= "<div class='content-image'><img src='https://static.igem.org/mediawiki/2013/thumb/4/42/Bonn_OutlookCCSspB1_Version2.png/726px-Bonn_OutlookCCSspB1_Version2.png'>sspB structure and its conservation among C. crescentus, E. coli and H. influenzae <sup><a href=#421>42.1</a></sup></div>The sspB&alpha; dimeric structure is stabilized by two &alpha;-helices in interaction, as part B of the figure above shows, each of them located at the N-terminus of either sspB&alpha; molecule. The subsequent parts of the protein form a domain consisting of two &beta;-sheet structures, together building up the ssrA binding site. An unstructured area at the C-terminus being referred to as the XB module forms the ClpX binding part of the protein. It is connected to the rest of the molecule via a linker domain. <sup><a href=#422>42.2</a></sup> </br>Chien et al. <sup><a href=#421>42.1</a></sup> compared crystal structures of C. crescentus sspB&alpha; and its E. coli and H. influenzae sspB orthologs, discovering that in sspB&alpha; the &alpha;-helices are significantly longer, more twisted and cover a larger cross section area than the other two sspB orthologs. Also considering that &beta;-sheets are rotated by around 20&deg; in comparison to E. coli and H. influenzae orthologs, this leads to an antiparallel orientation of the two ssrA tagged protein bound to the ssrA binding sites of an sspB&alpha; dimer in C. crescentus, while they are parallel in &gamma;-protobacterial sspB.</br></br><div class='content-image'><img src='https://static.igem.org/mediawiki/2013/6/6c/Bonn_OutlookCCSspB2.png' align=left>By measuring GFP fluorescence intensity, decrease of GFP-<sup>CC</sup>ssrA concentration (1) without sspB&alpha; added, (2) with mutated sspB&alpha;(Q74A) added , (3) with wildtype sspB&alpha; added can be visualized. <sup><a href=#421>42.1</a></sup></div>Chien et al. point out that although there are the remarkable differences in protein structure between sspB&alpha; and its &gamma;-protobacterial ortholog, they show up with similar effectiveness in binding proteins tagged with the related ssrA peptide. But it turned out in their research that effectiveness of sspB&alpha; binding to the protein which needs to be tethered to the ClpXP protease strongly depends on which ssrA ortholog the protein is tagged with. sspB&alpha; binds firmly to <sup>CC</sup>ssrA, with an affinity being 175 times as large as for binding to <sup>EC</sup>ssrA (i.e. the E. coli ortholog). By comparing the crystal structures of both sspB&alpha; and the compound of sspB&alpha; and <sup>CC</sup>ssrA, Chien et al. further proved that binding of sspB&alpha; to <sup>CC</sup>ssrA does not lead to significant changes of its 3D conformation.</br></br><h2>References</h2></br><a name=421>42.1</a> <a href='http://www.ncbi.nlm.nih.gov/pubmed/17937918'>Structure and substrate specificity of an SspB ortholog: design implications for AAA+ adaptors, Chien et al., Cell Press, 2007, PMID: 17937918</a></br><a name=422>42.2</a><a href='http://www.ncbi.nlm.nih.gov/pubmed/14967151'> Bivalent tethering of sspB to ClpXP is required for efficient substrate delivery: a protein design study, Bolon DN et al., Mol Cell, 2004, PMID: 14967151</a>";
@@ Line 389: / Line 391: @@
 content.parents=[40];
 content.childs=[];
-content.titleShort = "SspB Split";
+content.titleShort = "<sup>Cc</sup>sspB Split";
-content.titleLong = "SspB Split in C. crescentus";
+content.titleLong = "sspB Split in C. crescentus";
 content.summary= "The protein degradation system in Caulobacter crescentus resembles the system in E. coli, but the respective sequences of ssrA and SspB differ. Thus the different specifities can be used to introduce the ccSspB split system in wildtyp E. coli without disturbing the native processes in it.";
 content.text= "The protein degradation system in Caulobacter crescentus resembles the system in E. coli, but the respective sequences of ssrA and SspB differ <sup><a href=#431>43.1</a></sup>. Thus ccssrA only binds ccSspB but not E. coli SspB. <sup><a href=#432>43.2</a></sup> <sup><a href=#433>43.3</a></sup> <sup><a href=#434>43.4</a></sup> However, proteins tagged with ccssrA can be degraded by E. coli ClpXP. Therefore the utilization of ccSspB and ccssrA in E. coli has the advantage that SspB+ strains can be used. <sup><a href=#431>43.1</a></sup> </br> In order to use this with the SspB split system, the fusion proteins ccSspBΔ10-FRB and FKBP12-SspB[XB] (E. coli) were incubated with GFP-ccDAS+4 and E. coli ClpXP in vitro. Without rapamycin there was no degradation detected. Equally, addition of E. coli SspB showed no degradation.  Addition of rapamycin led to a reduction of GFP-ccDAS+4 of around 12% in 180 seconds. Compared to the E. coli split system (around 30 % in 180 seconds) this system is less fast but can, at least in vitro, be used with sspB-wildtype E. coli <sup><a href=#431>43.1</a></sup>. </br> As the results of the E.coli and the C. crescentus system in vitro show many similarities and the E. coli system works in vivo. It may be possible to use the C. crescentus in vivo as well.  </br> <img src='https://static.igem.org/mediawiki/2013/8/82/Bonn-ccSspB.jpg'> <sup><a href=#431>43.1</a></sup> <h2>References:</h2>  </br> <a name=431>43.1</a> <a href='http://dspace.mit.edu/bitstream/handle/1721.1/58089/654116495.pdf?sequence=1'> Understanding and Harnessing Energy-Dependent Proteolysis for Controlled Protein Degradation in Bacteria, J. Davis, Massachusetts Institute of Technology, april 2010  </a> </br> <a name=432>43.2</a> <a href='http://www.ncbi.nlm.nih.gov/pmc/articles/PMC58509/'> Overlapping recognition determinants within the ssrA degradation tag allow modulation of proteolysis, Flynn et al, Proc Natl Acad Sci USA 2001 Sep 11, PMID: 11535833 </a> </br> <a name=433>43.3</a> <a href='http://www.ncbi.nlm.nih.gov/pubmed/17937918'> Structure and substrate specifity of an SspB ortholog: design implications for AAA+ adaptors, Chien et al, Structure 2007 Oct, PMID: 17937918 <a/> </br> <a name=434>43.4</a> <a href='http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2581644/'> Inducible protein degradation in Bacillus subtilis using heterologous peptide tags and adaptor proteins to target substrates to the protease ClpXP, Griffith and Grossman, Mol Microbiol. 2008 Nov, PMID: 18811726 <a/> </br>";
@@ Line 401: / Line 403: @@
 content.parents=[74];
 content.childs=[];
-content.titleShort = "SsrA";
+content.titleShort = "<sup>Cc</sup>SsrA";
 content.titleLong = "C. crescentus ssrA and its application in E. coli";
 content.summary= "Proteins that need to be degraded by the ClpXP protease have to be tagged with the ssrA peptide previosly. Here is some information on the structure of ssrA.";
@@ Line 501: / Line 503: @@
 content.i =52;
 content.parents=[37];
-content.childs=[70];
+content.childs=[];
 content.titleShort="Achievements";
 content.titleLong="Achievements";
@@ Line 526: / Line 528: @@
 content.childs=[67, 68];
 content.titleShort = "Kill-switch for Lab Safety";
-content.titleLong = "Kill-switch systems using stress-induced toxin-antitoxin modules in Escherichia coli";
+content.titleLong = "Light inducible Kill-switch systems using toxin-antitoxin modules";
 content.summary= "Toxin-antitoxin systems are composed by an antitoxin and a toxin coding gene. Connecting our light inducible protein degradation system to the antitoxin via an ssrA-tag allows light induced cell death, as predominance of the toxin in a bacterium activates a cell death pathway.";
 content.text= "Our system of light inducible protein degradation can be utilized to degrade any specific protein and is thus usable in a light induced kill-switch system. For this application a connection between the degradation system and a toxin-antitoxin module like MazEF or ccdA/ccdB is needed. Either the toxin or the antitoxin could be light inducibly degraded by adding an ssrA-tag, which is detected by our degradation system, to its genetical code: <ul><li><b>Using the degradation of the toxin:</b> For that purpose the insertion of a plasmid containing the ssrA-tagged toxin encoding gene is needed. Since the predominance of the toxin activates a cell death pathway in bacteria, a bacterium containing a module that allows light inducible degradation of the toxin would only be viable, when the toxin is degraded. In darkness the toxin overexpression is no longer compensated and aggregation of it leads to cell death. Apart from the use in lab security such a kill-switch system would also be useful for environmental applications of bacteria, since it opens up the possibility of deploying bacteria for only one day and ensures their passing by nightfall. For example bacteria could be used to perform the ecological stabilization of a lake but after one night any genetically modified bacteria would be dead.</li><li><b>Using the degradation of the antitoxin:</b> Two plasmids are needed: The first one to express the toxin and the second one to express the ssra-tagged antitoxin, in such manner that the amounts of the toxin and the antitoxin are in equilibrium. Once light induces the degradation system, the antitoxin is degraded and the predominant toxin will kill the bacterium. </li></ul> Regarding our idea to improve lab security by implementing a kill-switch system, both described ways seem possible. Usage of the former would require cultivating and working with the bacteria under constant blue light, as darkness would kill them. Realization of the latter would require no usage of any blue light in the lab since bacteria which get into touch with daylight our any blue light would be killed. Due to the high light sensivity of our degradation system it can most likely be induced by daylight, which renders the former killswitch system useless. Bacteria which escape from the lab could survive simply through contact with daylight. Consequently we focused on the second system (the degradation of the antitoxin).</br> With MazEF we described a light inducible kill-switch system via the insertion of plasmids into bacteria. However a final kill-switch system would have to be introduced into the genomic DNA since plasmids in bacteria can be ejected, for instance via cell division, whereas a genomic DNA mutation is less likely to occur. Nevertheless the risk of a loss of function cannot be eliminated , which is why a secure system should countain much more than one kill-switch system to compensate the malfunction of a single kill-switch system. Therefore, we consider the MazEF kill-switch system to be part of a much larger security system for genetically engineered bacteria. </br>";
@@ Line 548: / Line 550: @@
 content.i =57;
 content.parents=[37];
-content.childs=[58,59,60,61,62,63,64];
+content.childs=[];
 content.titleShort = "Results";
 content.titleLong = "Results";
 content.summary= "Results";
-content.text="Results";
-content.type="Project";
-break;
-case 58:
-content.i =58;
-content.parents=[57];
-content.childs=[];
-content.titleShort = "Expression mCherry";
-content.titleLong = "Expression mCherry";
-content.summary= "";
-content.text="";
-content.type="Project";
-break;
-case 59:
-content.i =59;
-content.parents=[57];
-content.childs=[];
-content.titleShort = "mCherry Abbau";
-content.titleLong = "mCherry Abbau";
-content.summary= "";
-content.text="";
-content.type="Project";
-break;
-case 60:
-content.i =60;
-content.parents=[57];
-content.childs=[];
-content.titleShort = "Rapamycin Abbauinduktion";
-content.titleLong = "Rapamycin Abbauinduktion";
-content.summary= "";
-content.text="";
-content.type="Project";
-break;
-case 61:
-content.i =61;
-content.parents=[57];
-content.childs=[];
-content.titleShort = "pDawn";
-content.titleLong = "pDawn";
-content.summary= "";
-content.text="";
-content.type="Project";
-break;
-case 62:
-content.i =62;
-content.parents=[57];
-content.childs=[];
-content.titleShort = "ccdB killswitch";
-content.titleLong = "ccdB killswitch";
-content.summary= "";
-content.text="";
-content.type="Project";
-break;
-case 63:
-content.i =63;
-content.parents=[57];
-content.childs=[];
-content.titleShort = "MazF killswitch";
-content.titleLong = "MazF killswitch";
-content.summary= "";
-content.text="";
-content.type="Project";
-break;
-case 64:
-content.i =64;
-content.parents=[57];
-content.childs=[];
-content.titleShort = "LOV degradation induction";
-content.titleLong = "LOV degradation induction";
-content.summary= "";
 content.text="";
 content.type="Project";
@@ Line 637: / Line 562: @@
 content.parents=[37];
 content.childs=[];
-content.titleShort = "Generated plasmids";
+content.titleShort = "Plasmids";
-content.titleLong = "Generated plasmids";
+content.titleLong = "A list of generated plasmids";
-content.summary= "";
+content.summary= "A list of each and every plasmid we designed combined with short summaries of their function";
-content.text="";
+content.text="<groupparts></groupparts></br></br><a href='https://static.igem.org/mediawiki/2013/e/ef/Bonn_AraC-pBAD%28D%29-RBS32-mazF-TT_BBa_K1075040.jpg'><img src='https://static.igem.org/mediawiki/2013/e/ef/Bonn_AraC-pBAD%28D%29-RBS32-mazF-TT_BBa_K1075040.jpg' width='700px'></a>MazF is part of the biological toxin-antitoxin system MazEF. This system is derived from the bacterium Escherichia coli, where it serves as a stress inducable killswitch. MazF, a RNA-degrading enzyme, ist he toxin and is normally inhibited by the presence of MazE. Absence of MazE leads to cell death.<sup><a href=http://parts.igem.org/Part:BBa_K1075040>1</a></sup><a href='https://static.igem.org/mediawiki/2013/3/34/Bonn_pLac1-RBS32-mazF-TT_BBa_K1075039.jpg'><img src='https://static.igem.org/mediawiki/2013/3/34/Bonn_pLac1-RBS32-mazF-TT_BBa_K1075039.jpg' width='700px'></a>We used this part to gain an IPTG and Lactose inducible construct of MazF. This part was used to test the toxicity of MazF as part of our proof of concept. <sup><a href=http://parts.igem.org/Part:BBa_K1075039>2</a></sup><a href='https://static.igem.org/mediawiki/2013/8/87/Bonn_pLac2-RBS32-mazE-%28ec%29ssrA%28DAS%2B4%29-TT_BBa_K1075043.jpg'><img src='https://static.igem.org/mediawiki/2013/8/87/Bonn_pLac2-RBS32-mazE-%28ec%29ssrA%28DAS%2B4%29-TT_BBa_K1075043.jpg' width='700px'></a>This part can be used to gain a Lactose and IPTG inducable construct of mazE-(ec)ssrA(DAS+4). We used this construct as part of our proof of concept. <sup><a href=http://parts.igem.org/Part:BBa_K1075043>3</a></sup><a href='https://static.igem.org/mediawiki/2013/c/cf/Bonn_AraC-pBAD-RBS-mazF-TT-pLac2-RBS-mazE-%28ec%29ssrA%28DAS%2B4%29-TT_BBa_K1075048.jpg'><img src='https://static.igem.org/mediawiki/2013/c/cf/Bonn_AraC-pBAD-RBS-mazF-TT-pLac2-RBS-mazE-%28ec%29ssrA%28DAS%2B4%29-TT_BBa_K1075048.jpg' width='700px'></a>This construct can be used to gain IPTG and Lactose inducible expression of mazE(ec)ssrA(DAS+4) and arabinose inducible expression of MazF. Thus, both parts of the toxin-antitoxin system mazEF are inducible and this construct can be used to further examine this system. We used this construct as part of our proof of concept.<sup><a href=http://parts.igem.org/Part:BBa_K1075048>4</a></sup><a href='https://static.igem.org/mediawiki/2013/7/7f/Bonn_AraC-pBAD-RBS-sspB-TT-pLac2-RBS-mCherry-TT_BBa_K1075049.jpg'><img src='https://static.igem.org/mediawiki/2013/7/7f/Bonn_AraC-pBAD-RBS-sspB-TT-pLac2-RBS-mCherry-TT_BBa_K1075049.jpg' width='700px'></a>The sspB protein is an adaptor responsible for delivering ssrA-tagged substrates to the ClpXP protease in order to enhance their degradation. mCherry is a red fluorescent protein with the excitation maximum at 587 nm and the Emission maximum at 610 nm.<sup><a href=http://parts.igem.org/Part:BBa_K1075049>5</a></sup><a href='https://static.igem.org/mediawiki/2013/4/42/Bonn_pDawn_%28Prom_%28const.%29-RBS34-YF1-FixJ-FixK-LambdaC-pC%29_BBa_K1075044.jpg'><img src='https://static.igem.org/mediawiki/2013/4/42/Bonn_pDawn_%28Prom_%28const.%29-RBS34-YF1-FixJ-FixK-LambdaC-pC%29_BBa_K1075044.jpg' width='700px'></a>The plasmid pDawn was designed by Ohlendorf et al. in 2012 together with its counter plasmid pDusk. Both plasmids are single plasmid systems, which allow the activation (pDawn) or repression (pDusk) of gene expression by blue light. They are easy to implement in the laboratory and lead to up to 460-fold activity change upon ilumination. <sup><a href=http://parts.igem.org/Part:BBa_K1075044>6</a></sup><a href='https://static.igem.org/mediawiki/2013/5/50/Bonn_pDawn_%28Prom_%28const.%29-RBS34-YF1-FixJ-FixK-LambdaC-pC%29-RBS32-ccdB-TT_BBa_K1075047.jpg'><img src='https://static.igem.org/mediawiki/2013/5/50/Bonn_pDawn_%28Prom_%28const.%29-RBS34-YF1-FixJ-FixK-LambdaC-pC%29-RBS32-ccdB-TT_BBa_K1075047.jpg' width='700px'></a>pDawn-RBS32-ccdB-TT can be used to photocontrol the expression of the toxin ccdB. In this system pDawn (BBa_K1075044) can be regulated by blue light, while ccdB (BBa_K1075029) acts as a killswitch. The ccd operon of the F plasmid encodes CcdB, a toxin targeting the essential gyrase of Escherichia coli, and CcdA, the unstable antidote that interacts with CcdB to neutralize its toxicity.<sup><a href=http://parts.igem.org/Part:BBa_K1075047>7</a></sup><a href='https://static.igem.org/mediawiki/2013/0/03/Bonn_AraC-pBAD%28D%29-RBS32-SspB%28Core%28-LOV-ipaA-TT_BBa_K1075019.jpg'><img src='https://static.igem.org/mediawiki/2013/0/03/Bonn_AraC-pBAD%28D%29-RBS32-SspB%28Core%28-LOV-ipaA-TT_BBa_K1075019.jpg' width='700px'></a>The part can be used to regulate the dimerization of the two parts of the split sspB. The binding of ipaA to Vincolin is regulated via the light-sensitive LOV domain. Therefore the two sspB parts only come together when the LOV domain was activated with blue light. <sup><a href=http://parts.igem.org/Part:BBa_K1075019>8</a></sup><a href='https://static.igem.org/mediawiki/2013/d/d0/Bonn_J23105-RBS32-SspB%28Core%29-LOV-ipaA-TT_BBa_K1075018.jpg'><img src='https://static.igem.org/mediawiki/2013/d/d0/Bonn_J23105-RBS32-SspB%28Core%29-LOV-ipaA-TT_BBa_K1075018.jpg' width='700px'></a>The part can be used to regulate the dimerization of the two parts of the split sspB. The binding of ipaA to Vincolin is regulated via the light-sensitive LOV domain. Therefore the two sspB parts only come together when the LOV domain was activated with blue light. <sup><a href=http://parts.igem.org/Part:BBa_K1075018>9</a></sup><a href='https://static.igem.org/mediawiki/2013/5/5a/Bonn_AraC-pBAD%28D%29-RBS32-VinD1-sspBXB-TT_BBa_K1075016.jpg'><img src='https://static.igem.org/mediawiki/2013/5/5a/Bonn_AraC-pBAD%28D%29-RBS32-VinD1-sspBXB-TT_BBa_K1075016.jpg' width='700px'></a>The part can be used to regulate the dimerization of the two parts of the split sspB. As Vincolin bins to ipaA, the sspB dimer is stabilized. We used this part to achieve a light-induced dimerization of the sspB parts.  <sup><a href=http://parts.igem.org/Part:BBa_K1075016>10</a></sup><a href='https://static.igem.org/mediawiki/2013/9/9c/Bonn_J23118-RBS32-VinD1-SspBXB-TT_BBa_K1075015.jpg'><img src='https://static.igem.org/mediawiki/2013/9/9c/Bonn_J23118-RBS32-VinD1-SspBXB-TT_BBa_K1075015.jpg' width='700px'></a>The part can be used to regulate the dimerization of the two parts of the split sspB. As Vincolin bins to ipaA, the sspB dimer is stabilized. We used this part to achieve a light-induced dimerization of the sspB parts. <sup><a href=http://parts.igem.org/Part:BBa_K1075015>11</a></sup><a href='https://static.igem.org/mediawiki/2013/7/78/Bonn_pLac_pBAD2-RBS32-SspB-TT_BBa_K1075006.jpg'><img src='https://static.igem.org/mediawiki/2013/7/78/Bonn_pLac_pBAD2-RBS32-SspB-TT_BBa_K1075006.jpg' width='700px'></a>The sspB protein is an adaptor responsible for delivering ssrA-tagged substrates to the ClpXP protease in order to enhance their degradation. Prefixed is RBS, the binding site for a ribosome and the pLac promoter. It also includes a double terminator 'TT', which interrupts the translation. <sup><a href=http://parts.igem.org/Part:BBa_K1075006>12</a></sup><a href='https://static.igem.org/mediawiki/2013/8/8f/Bonn_E._coli_ssrA%28DAS%2B4%29-TT_BBa_K1075021.jpg'><img src='https://static.igem.org/mediawiki/2013/8/8f/Bonn_E._coli_ssrA%28DAS%2B4%29-TT_BBa_K1075021.jpg' width='700px'></a>The (ec)ssrA(DAS+4) tag can be fused to proteins so that they will be delivered to the ClpXP protease and degraded.<sup><a href=http://parts.igem.org/Part:BBa_K1075021>13</a></sup><a href='https://static.igem.org/mediawiki/2013/6/63/Bonn_AraC-pBad-RBS34-mCherry-ecssrA%28DAS%2B4%29-TT_BBa_K1075025.jpg'><img src='https://static.igem.org/mediawiki/2013/6/63/Bonn_AraC-pBad-RBS34-mCherry-ecssrA%28DAS%2B4%29-TT_BBa_K1075025.jpg' width='700px'></a>As we want to control protein degradation by controlling the function of ecSspB, we tagged the red fluorescent protein mCherry with ecssrA(DAS+4) to measure the degradation rate. The part was designed for proof of principle. Application as a bacterial fotographic film might be possible as well. <sup><a href=http://parts.igem.org/Part:BBa_K1075025>14</a></sup><a href='https://static.igem.org/mediawiki/2013/6/63/Bonn_AraC-pBad-RBS34-mCherry-ecssrA%28DAS%2B4%29-TT_BBa_K1075025.jpg'><img src='https://static.igem.org/mediawiki/2013/6/63/Bonn_AraC-pBad-RBS34-mCherry-ecssrA%28DAS%2B4%29-TT_BBa_K1075025.jpg' width='700px'></a>As we want to control protein degradation by controlling the function of ecSspB, we tagged the red fluorescent protein mCherry with ecssrA(DAS+4) to measure the degradation rate. The part was designed for proof of principle. Application as a bacterial fotographic film might be possible as well. <sup><a href=http://parts.igem.org/Part:BBa_K1075025>15</a></sup><a href='https://static.igem.org/mediawiki/2013/c/cd/Bonn_pJD427%28pC-RBS32-FKBP12-EcsspB%28XB%29-Term-pB-RBS32-EcsspB%28Core%29-FRB-Term-Term%29_BBa_K1075010.jpg'><img src='https://static.igem.org/mediawiki/2013/c/cd/Bonn_pJD427%28pC-RBS32-FKBP12-EcsspB%28XB%29-Term-pB-RBS32-EcsspB%28Core%29-FRB-Term-Term%29_BBa_K1075010.jpg' width='700px'></a>The SspB protein is an adaptor responsible for delivering ssrA-tagged substrates to the ClpXP protease in order to enhance their degradation. To control degradation it is reasonable to control the function of SspB. That is made by splitting it into two parts, each of which cannot induce degradation on its own. To bring both SspB parts together again for inducible degradation, they were combined with a chemical inducible heterodimerisation system: FRB and FKBP12. These two parts interact in the presence of rapamycin <sup><a href=http://parts.igem.org/Part:BBa_K1075010>16</a></sup><a href='https://static.igem.org/mediawiki/2013/d/d6/Bonn_%28pJD427%29-pLac-RBS34-mCherry-ecssrA%28DAS%2B4%29-TT_BBa_K1075027.jpg'><img src='https://static.igem.org/mediawiki/2013/d/d6/Bonn_%28pJD427%29-pLac-RBS34-mCherry-ecssrA%28DAS%2B4%29-TT_BBa_K1075027.jpg' width='700px'></a>As we want to control protein degradation by controlling the function of ecSspB, we tag the red fluorescent protein mCherry with ecssrA(DAS+4) to measure the degradation rate. We control ecSspB by splitting it into two parts each of which cannot induce degradation on its own but regains function by addition of rapamycin. As the co-transformation of the two plasmids didn’t work, we combined them to one.<sup><a href=http://parts.igem.org/Part:BBa_K1075027>17</a></sup><a href='https://static.igem.org/mediawiki/2013/b/bc/Bonn_pJD427-pLac-RBS34-mCherry-TT_BBa_K1075028.jpg'><img src='https://static.igem.org/mediawiki/2013/b/bc/Bonn_pJD427-pLac-RBS34-mCherry-TT_BBa_K1075028.jpg' width='700px'></a>This plasmid is a negative control to the pJD427-pLac2-RBS34-mCherry-ecssrA(DAS+4)-TT plasmid. <sup><a href=http://parts.igem.org/Part:BBa_K1075028>18</a></sup><a href='https://static.igem.org/mediawiki/2013/c/cd/Bonn_pJD427%28pC-RBS32-FKBP12-EcsspB%28XB%29-Term-pB-RBS32-EcsspB%28Core%29-FRB-Term-Term%29_BBa_K1075010.jpg'><img src='https://static.igem.org/mediawiki/2013/c/cd/Bonn_pJD427%28pC-RBS32-FKBP12-EcsspB%28XB%29-Term-pB-RBS32-EcsspB%28Core%29-FRB-Term-Term%29_BBa_K1075010.jpg' width='700px'></a>The Plasmid pJD427 contains the fusion proteins SspB[CORE]-FRB and FKBP12-SspB[XB]: SspB[CORE]-FRB with the weak constitutive promoter proB, FKBP12-SspB[XB] with the strong constitutive promoter proC and a medium-copy p15a origin of replication.  <sup><a href=http://parts.igem.org/Part:BBa_K1075010>19</a></sup><a href='https://static.igem.org/mediawiki/2013/e/e1/Bonn_pLac-RBS32-ccdA-ssrA-TT_BBa_K1075035.jpg'><img src='https://static.igem.org/mediawiki/2013/e/e1/Bonn_pLac-RBS32-ccdA-ssrA-TT_BBa_K1075035.jpg' width='700px'></a>CcdA inhibits the toxin ccdB by binding to it and thus represses cell death. The part contains ccdA with a ssrA tag under the control of the Lac operon. <sup><a href=http://parts.igem.org/Part:BBa_K1075035>20</a></sup><a href='https://static.igem.org/mediawiki/2013/8/81/Bonn_AraC-pBAD%28D%29-RBS32-ccdB-TT_BBa_K1075031.jpg'><img src='https://static.igem.org/mediawiki/2013/8/81/Bonn_AraC-pBAD%28D%29-RBS32-ccdB-TT_BBa_K1075031.jpg' width='700px'></a>The part contains a chemically induced kill-switch. When Arabinose is added to the bacteria, the toxin ccdB is expressed and cell death is initiated. <sup><a href=http://parts.igem.org/Part:BBa_K1075031>20</a></sup>";
 content.type="Project";
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 content.childs=[];
 content.titleShort = "MazEF";
-content.titleLong = "A kill-switch system using the stress-induced toxin-antitoxin module MazEF in Escherichia coli";
+content.titleLong = "The maz toxin-antitoxin system";
 content.summary= "The toxin-antitoxin system MazEF is composed by an upstream gene <i>mazE</i>, encoding a labile antitoxin, and a downstream gene <i>mazF</i>, that encodes a stable toxin. Connecting our light inducible protein degradation system to the antitoxin MazE allows light inducible cell death, as predominance of MazF in a bacterium activates cell death pathway.";
-content.text= "Our system of light inducible protein degradation can be utilized to degrade any specific protein and is therefore usable to realize a light inducible kill-switch system. A connection of protein degradation to a cell death pathway is represented by the stress-induced toxin-antitoxin module <i>mazEF</i> in Escherichia coli. <i>mazEF</i> is located on the chromosome in E.coli which is associated with programmed cell death. The toxin-antitoxin system is composed by an upstream gene <i>mazE</i>, encoding a labile antitoxin, and a downstream gene mazF, that encodes a stable toxin. </br>The product of <i>mazF</i> cleaves mRNAs and tmRNAs at a specific site, which leads to an inhibition of translation. MazF shows a specific cleaving mechanism, which is not well understood yet, but shows that there is also protein synthesis which is unaffected by MazF. These proteins are presumably part of a cell death pathway. </br>The effect of <i>mazF</i> is suppressed by the Product of <i>mazE</i> which is degraded by the Protease ClpAP in bacteria. As a result of stressful conditions expression of the chromosomal <i>mazEF</i> module is reduced which leads to an imbalance between the products of <i>mazF</i> and <i>mazE</i>: When expression is lowered the stable toxin of <i>mazF</i> still persists while the labile antitoxin of <i>mazE</i> is degraded and can no longer suppress the effect of <i>mazF</i> leading to acute toxicity and cell death. </br><i>MazEF</i>-mediated cell death in E. coli can be caused by:<ul><li>extreme amino acid starvation<sup><a href = #671>67.1</a></sup></li><li> inhibition of transcription and/or translation by antibiotics such as rifampin, chloramphenicol, and spectinomycin under specific growth conditions<sup><a href = #671>67.1</a></sup></li><li>inhibition of translation by the Doc protein of prophage P1<sup><a href = #671>67.1</a></sup></li><li>DNA damage caused by thymine starvation as well as by mitomycin C, nalidixic acid, and UV irradiation<sup><a href = #671>67.1</a></sup></li><li>oxidative stress (H2O2)<sup><a href = #671'>67.1</a></sup></li></ul>Amitai et al. tested in 2004 the Hypothesis of Pedersen et al.<sup><a href = #672>67.2</a></sup>, that chromosomal toxin-antitoxin systems may rather cause a state of reversible bacteriostasis than programmed cell death<sup><a href = #671>67.1</a></sup>.Therefore E.coli strain MC4100 &#916<i>mazE</i>F relA1 lacIq was cotransformated with:<ul><li>pBad-<i>mazF</i></li><li>pQE-&#916his-<i>mazE</i></li></ul><i>mazF</i>-expression can be induced by the addition of Arabinose via the pBad promoter of the first plasmid. The transformation of the second plasmid results firstly in the repression of <i>mazE</i> expression, whereas when IPTG is added <i>mazE</i> production is induced.</br><div class='content-image' align='center' height=501 width=410><a href='https://static.igem.org/mediawiki/2013/a/ac/Team_Bonn_MazF_1.png'><img src='https://static.igem.org/mediawiki/2013/a/ac/Team_Bonn_MazF_1.png' height=491 width=400></a></br><i>Ability of E. coli cells that had been ectopically overexpressing MazF in liquid medium to form colonies when ectopically overexpressing MazE on plates. The cultures were grown in LB medium (A) or M9 minimal medium with 0.5% glycerol (B) at 37°C to midlogarithmic phase (OD600, 0.5)<sup><a href = #671>67.1</a></sup>.</i></div>Using these tools, Amitai et al. tested the effect of MazE overproduction on MazF-overproducing bacteria during growth in liquid medium.</br>The E.coli strain was incubated in LB medium. After <i>mazF</i> expression was induced by adding arabinose two samples were taken at several time points. To repress <i>mazF</i> expression to both of them glucose was added. In addition IPTG was added to one culture to induce <i>mazE</i> expression. The two cultures were compared via the level of protein synthesis and OD600.</br>Finally Amitai et al. confirmed the assumption that the overproduction of MazE after until 6h under overproduction of MazF could resuscitate E.coli cells in LB medium (Fig. 1A)<sup><a href = #671>67.1</a></sup>, but the longer MazF was induced the less cells could be resuscitated by MazE.</br>Whereas MazE overproduction can reverse the inhibitory effect of MazF on translation, it cannot reverse the effect of MazF on colony formation, which is shown in figure 2. Only 1h after the induction of MazE expression, the rate of translation was restored to nearly 100% (Fig.2 Aa, Ab, Ac) but the bacteriocidic effect could not be reversed (Fig.2 Ba, Bb, Bc).</br>Additionally, Amtai et al. found out, that in M9 medium MazE was less able to reverse the effects of MazF overexpression than in LB medium (Fig.1B vs. 1A). it was concluded that there is a point of no return, when MazE is inable to resuscitate a MazF damaged cell, which occurs earlier in M9 medium than in LB medium.</br>Based on their results a model of the MazEF mechanism was built: A <i>mazF</i>-mediated cascade leads to a cell death pathway, but can nevertheless be stopped at several intermediary steps by e.g. <i>mazE</i>. When a point of no return is reached, the cascade cannot be stopped anymore.<div class='content-image' align='center' height=827 width=784><a href='https://static.igem.org/mediawiki/2013/9/9e/Team_Bonn_MazF_2.png'><img src='https://static.igem.org/mediawiki/2013/9/9e/Team_Bonn_MazF_2.png' height=817 width=764></a></br><i>Effect of MazE overproduction during growth in liquid medium on the ability of MazF-overproducing E. coli cells to synthesize proteins and form colonies. To induce <i>mazE</i> expression, IPTG was added to the bacterial culture at 1 h (Aa and Ba), 4 h (Ab and Bb), and 6 h (Ac and Bc) after <i>mazF</i> induction at time zero. The effects of the ectopic overexpression of MazE were measured at 1 and 3 h after the induction of <i>mazE</i> expression.<sup><a href = #671>67.1</a></sup>.</i></div></br>Back to our project and to the idea of a light inducible kill-switch system:</br>As we described in the previous paragraph for both of our kill-switch systems using the MazEF module either MazF or MazE could be degraded:<ul><li>Using the degradation of MazF:</b> For that purpose the insertion of a plasmid containing the ssrA-tagged toxin encoding gene is needed. Since the predominance of the toxin activates a cell death pathway in bacteria, a bacterium containing a module that allows light inducible degradation of the MazF would only be viable, when it is degraded. In darkness the MazF overexpression is no longer compensated and aggregation of it leads to cell death. </li><li><b>Using the degradation of the antitoxin:</b> Two plasmids are needed: The first one to express the MazF and the second one to express the ssra-tagged MazE, in such manner that the amounts of the <i>MazF</i> and the <i>MazE</i> are in equilibrium. Once light induces the degradation system, the MazE is degraded and the predominant toxin will kill the bacterium.</li></ul>As we explained in the general kill-switch system text we finally focused on the second system (via the degradation of MazE).</br>With the design of a MazEF kill-switch system the possibility of resuscitating bacteria in the way Amitai et al. showed has to be considered. A predominant MazF could kill a bacterium in LB within about two hours, but it needs to be predominant over a long period (>7h) to induce its death without it being resuscitated by renewed <i>mazE</i> expression (Fig.1A)with more than 50% probability .</br>Certainly these facts seem to be unfavourable for the realization of a kill switch system via MazEF, but fortunately our system of heterodimerization (Lungu et al.) allows long continuous degradation<sup><a href = #673>67.3</a></sup>, due to the high stability of the light induced hererodimer. Therefore likely a short exposure time will result in prolonged protein degradation sufficing for bacterial death. Additionally, Amitai et al. showed that the less nutrition is available for a bacterium, the earlier the point of no return is reached. If a bacterium escapes the lab, it will likely have less nutrition available than in LB medium. It might reach the point of no return earlier.</br>We described a light inducible MazEF kill-switch system via the insertion of plasmids into bacteria. However a final kill-switch system would have to be implemented in the genomic DNA since plasmids in bacteria can be ejected, for instance via cell division, whereas a genomic DNA mutation is less likely to occur. Nevertheless risk of a loss of function cannot be eliminated , which is why a secure system should countain much more than one kill-switch system to compensate the malfunction of a single kill-switch system. Therefore, we consider the MazEF kill-switch system to be part of a much larger security system for genetically engineered bacteria.<h2>References</h2><a name =671>67.1</a> <a href = 'http://www.ncbi.nlm.nih.gov/pmc/articles/PMC532418/'>MazF-Mediated Cell Death in Escherichia coli: a Point of No Return, Shahar Amitai et al., Journal of Bacteriology Vol. 186, No. 24, 2004, p.8295–8300.</a></br><a name =672>67.2</a> <a href = 'http://www.ncbi.nlm.nih.gov/pubmed/?term=Rapid+induction+and+reversal+of+bacteriostatic+conditions+by+controlled+expression+of+toxins+and+antitoxins'>Rapid induction and reversal of bacteriostatic conditions by controlled expression of toxins and antitoxins, Pedersen et al., Molecular Microbiology 45, 2002, 501–510.</a></br><a name =673>67.3</a> <a href = 'http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3334866/'>Designing Photoswitchable Peptides Using the AsLOV2 Domain, Oana I. Lungu et al., Chem Biol. 2012, 19(4):507-17.</a>";
+content.text= "Our system of light inducible protein degradation can be utilized to degrade any specific protein and is therefore usable to realize a light inducible kill-switch system. A connection of protein degradation to a cell death pathway is represented by the stress-induced toxin-antitoxin module <i>mazEF</i> in Escherichia coli. <i>mazEF</i> is located on the chromosome in E.coli which is associated with programmed cell death. The toxin-antitoxin system is composed by an upstream gene <i>mazE</i>, encoding a labile antitoxin, and a downstream gene mazF, that encodes a stable toxin. </br>The product of <i>mazF</i> cleaves mRNAs and tmRNAs at a specific site, which leads to an inhibition of translation. MazF shows a specific cleaving mechanism, which is not well understood yet, but shows that there is also protein synthesis which is unaffected by MazF. These proteins are presumably part of a cell death pathway. </br>The effect of <i>mazF</i> is suppressed by the Product of <i>mazE</i> which is degraded by the Protease ClpAP in bacteria. As a result of stressful conditions expression of the chromosomal <i>mazEF</i> module is reduced which leads to an imbalance between the products of <i>mazF</i> and <i>mazE</i>: When expression is lowered the stable toxin of <i>mazF</i> still persists while the labile antitoxin of <i>mazE</i> is degraded and can no longer suppress the effect of <i>mazF</i> leading to acute toxicity and cell death. </br><i>MazEF</i>-mediated cell death in E. coli can be caused by:<ul><li>extreme amino acid starvation<sup><a href = #674>67.4</a></sup><sup><a href = #675>67.5</a></sup></li><li> inhibition of transcription and/or translation by antibiotics such as rifampin, chloramphenicol, and spectinomycin under specific growth conditions<sup><a href = #676>67.6</a></sup></li><li>inhibition of translation by the Doc protein of prophage P1<sup><a href = #677>67.7</a></sup></li><li>DNA damage caused by thymine starvation<sup><a href = #678>67.8</a></sup> as well as by mitomycin C, nalidixic acid, and UV irradiation<sup><a href = #679>67.9</a></sup></li><li>oxidative stress (H2O2)<sup><a href = #679'>67.9</a></sup></li></ul>Amitai et al. tested in 2004 the Hypothesis of Pedersen et al.<sup><a href = #672>67.2</a></sup>, that chromosomal toxin-antitoxin systems may rather cause a state of reversible bacteriostasis than programmed cell death<sup><a href = #671>67.1</a></sup>.Therefore E.coli strain MC4100 &#916<i>mazE</i>F relA1 lacIq was cotransformated with:<ul><li>pBad-<i>mazF</i></li><li>pQE-&#916his-<i>mazE</i></li></ul><i>mazF</i>-expression can be induced by the addition of Arabinose via the pBad promoter of the first plasmid. The transformation of the second plasmid results firstly in the repression of <i>mazE</i> expression, whereas when IPTG is added <i>mazE</i> production is induced.</br><div class='content-image' align='center' height=501 width=410><a href='https://static.igem.org/mediawiki/2013/a/ac/Team_Bonn_MazF_1.png'><img src='https://static.igem.org/mediawiki/2013/a/ac/Team_Bonn_MazF_1.png' height=491 width=400></a></br><i>Ability of E. coli cells that had been ectopically overexpressing MazF in liquid medium to form colonies when ectopically overexpressing MazE on plates. The cultures were grown in LB medium (A) or M9 minimal medium with 0.5% glycerol (B) at 37°C to midlogarithmic phase (OD600, 0.5)<sup><a href = #671>67.1</a></sup>.</i></div>Using these tools, Amitai et al. tested the effect of MazE overproduction on MazF-overproducing bacteria during growth in liquid medium.</br>The E.coli strain was incubated in LB medium. After <i>mazF</i> expression was induced by adding arabinose two samples were taken at several time points. To repress <i>mazF</i> expression to both of them glucose was added. In addition IPTG was added to one culture to induce <i>mazE</i> expression. The two cultures were compared via the level of protein synthesis and OD600.</br>Finally Amitai et al. confirmed the assumption that the overproduction of MazE after until 6h under overproduction of MazF could resuscitate E.coli cells in LB medium (Fig. 1A)<sup><a href = #671>67.1</a></sup>, but the longer MazF was induced the less cells could be resuscitated by MazE.</br>Whereas MazE overproduction can reverse the inhibitory effect of MazF on translation, it cannot reverse the effect of MazF on colony formation, which is shown in figure 2. Only 1h after the induction of MazE expression, the rate of translation was restored to nearly 100% (Fig.2 Aa, Ab, Ac) but the bacteriocidic effect could not be reversed (Fig.2 Ba, Bb, Bc).</br>Additionally, Amtai et al. found out, that in M9 medium MazE was less able to reverse the effects of MazF overexpression than in LB medium (Fig.1B vs. 1A). it was concluded that there is a point of no return, when MazE is inable to resuscitate a MazF damaged cell, which occurs earlier in M9 medium than in LB medium.</br>Based on their results a model of the MazEF mechanism was built: A <i>mazF</i>-mediated cascade leads to a cell death pathway, but can nevertheless be stopped at several intermediary steps by e.g. <i>mazE</i>. When a point of no return is reached, the cascade cannot be stopped anymore.<div class='content-image' align='center' height=827 width=784><a href='https://static.igem.org/mediawiki/2013/9/9e/Team_Bonn_MazF_2.png'><img src='https://static.igem.org/mediawiki/2013/9/9e/Team_Bonn_MazF_2.png' height=817 width=764></a></br><i>Effect of MazE overproduction during growth in liquid medium on the ability of MazF-overproducing E. coli cells to synthesize proteins and form colonies. To induce <i>mazE</i> expression, IPTG was added to the bacterial culture at 1 h (Aa and Ba), 4 h (Ab and Bb), and 6 h (Ac and Bc) after <i>mazF</i> induction at time zero. The effects of the ectopic overexpression of MazE were measured at 1 and 3 h after the induction of <i>mazE</i> expression.<sup><a href = #671>67.1</a></sup>.</i></div></br>Back to our project and to the idea of a light inducible kill-switch system:</br>As we described in the previous paragraph for both of our kill-switch systems using the MazEF module either MazF or MazE could be degraded:<ul><li>Using the degradation of MazF:</b> For that purpose the insertion of a plasmid containing the ssrA-tagged toxin encoding gene is needed. Since the predominance of the toxin activates a cell death pathway in bacteria, a bacterium containing a module that allows light inducible degradation of the MazF would only be viable, when it is degraded. In darkness the MazF overexpression is no longer compensated and aggregation of it leads to cell death. </li><li><b>Using the degradation of the antitoxin:</b> Two plasmids are needed: The first one to express the MazF and the second one to express the ssra-tagged MazE, in such manner that the amounts of the <i>MazF</i> and the <i>MazE</i> are in equilibrium. Once light induces the degradation system, the MazE is degraded and the predominant toxin will kill the bacterium.</li></ul>As we explained in the general kill-switch system text we finally focused on the second system (via the degradation of MazE).</br>With the design of a MazEF kill-switch system the possibility of resuscitating bacteria in the way Amitai et al. showed has to be considered. A predominant MazF could kill a bacterium in LB within about two hours, but it needs to be predominant over a long period (>7h) to induce its death without it being resuscitated by renewed <i>mazE</i> expression (Fig.1A)with more than 50% probability .</br>Certainly these facts seem to be unfavourable for the realization of a kill switch system via MazEF, but fortunately our system of heterodimerization (Lungu et al.) allows long continuous degradation<sup><a href = #673>67.3</a></sup>, due to the high stability of the light induced hererodimer. Therefore likely a short exposure time will result in prolonged protein degradation sufficing for bacterial death. Additionally, Amitai et al. showed that the less nutrition is available for a bacterium, the earlier the point of no return is reached. If a bacterium escapes the lab, it will likely have less nutrition available than in LB medium. It might reach the point of no return earlier.</br>We described a light inducible MazEF kill-switch system via the insertion of plasmids into bacteria. However a final kill-switch system would have to be implemented in the genomic DNA since plasmids in bacteria can be ejected, for instance via cell division, whereas a genomic DNA mutation is less likely to occur. Nevertheless risk of a loss of function cannot be eliminated , which is why a secure system should countain much more than one kill-switch system to compensate the malfunction of a single kill-switch system. Therefore, we consider the MazEF kill-switch system to be part of a much larger security system for genetically engineered bacteria.<h2>References</h2><a name =671>67.1</a> <a href = 'http://www.ncbi.nlm.nih.gov/pmc/articles/PMC532418/'>MazF-Mediated Cell Death in Escherichia coli: a Point of No Return, Shahar Amitai et al., Journal of Bacteriology Vol. 186, No. 24, 2004, p.8295–8300.</a></br><a name =672>67.2</a> <a href = 'http://www.ncbi.nlm.nih.gov/pubmed/?term=Rapid+induction+and+reversal+of+bacteriostatic+conditions+by+controlled+expression+of+toxins+and+antitoxins'>Rapid induction and reversal of bacteriostatic conditions by controlled expression of toxins and antitoxins, Pedersen et al., Molecular Microbiology 45, 2002, 501–510.</a></br><a name =673>67.3</a> <a href = 'http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3334866/'>Designing Photoswitchable Peptides Using the AsLOV2 Domain, Oana I. Lungu et al., Chem Biol. 2012, 19(4):507-17.</a></br><a name =674>67.4</a> <a href = 'http://www.ncbi.nlm.nih.gov/pmc/articles/PMC39188/'>An Escherichia coli chromosomal addiction module regulated by guanosine 3,5-bispyrophosphate: a model for programmed bacterial cell death, Aizenman, E., H. Engelberg-Kulka, and G. Glaser, Proc. Natl. Acad. Sci., 1996, USA 93:6059-6063.</a></br><a name =675>67.5</a> <a href = 'http://www.ncbi.nlm.nih.gov/pmc/articles/PMC28068/'>rexB of bacteriophage lambda is an anti-cell death gene. Engelberg-Kulka, H., M. Reches, S. Narasimhan, R. Schoulaker-Schwarz, Y. Klemes, E. Aizenman, and G. Glaser, Proc. Natl. Acad. Sci., 1998, USA 95:15481-15486.</a></br><a name =676>67.6</a> <a href = 'http://www.ncbi.nlm.nih.gov/pmc/articles/PMC95100/'>Programmed cell death in Escherichia coli: some antibiotics can trigger mazEF lethality, Sat, B., R. Hazan, T. Fisher, H. Khaner, G. Glaser, and H. Engelberg-Kulka, J. Bacteriol. 2001, 183:2041-2045.</a></br><a name =677>67.7</a> <a href = 'http://www.ncbi.nlm.nih.gov/pmc/articles/PMC95101/'>Postsegregational killing mediated by the P1 phage addiction module Phd-Doc requires the Escherichia coli programmed cell death system mazEF, Hazan, R., B. Sat, M. Reches, and H. Engelberg-Kulka, J. Bacteriol. 2001, 183:2046–2050.</a></br><a name =678>67.8</a> <a href = 'http://www.ncbi.nlm.nih.gov/pmc/articles/PMC150121/'>The Escherichia coli mazEF suicide module mediates thymineless death, Sat, B., M. Reches, and H. Engelberg-Kulka, J. Bacteriol. 2003, 185:1803–1807.</a></br><a name =679>67.9</a> <a href = 'http://www.ncbi.nlm.nih.gov/pmc/articles/PMC415763/'>Escherichia coli mazEFmediated cell death is triggered by various stressful conditions, Hazan, R., B. Sat, and H. Engelberg-Kulka, J. Bacteriol. 2004, 186:3663–3669.</a>";
 content.type="Project";
 break;
 case 68:
@@ Line 673: / Line 599: @@
 content.parents=[54];
 content.childs=[];
-content.titleShort = "ccdA/ccdB";
+content.titleShort = "ccdAB";
 content.titleLong = "The ccd toxin-antitoxin system";
-content.summary= "this is a summary6";
+content.summary= "CcdB leads to double strand breaks in bacterial genomic DNA and thus is a potent toxin leading to cell death. ccdA compromises the toxic effect of ccdB.";
 content.text= "The ccd module is a toxin-antitoxin (TA) system similar to the mazE/mazF system. The module is located on the F Plasmid in Escherichia coli bacteria and essential for their survival. Normally the toxin ccdB is inactivated by the presence of the antitoxin ccdA in the form of a ccdAB complex. If ccdA is no longer available, ccdB inhibits DNA gyrase which leads to cell death. Gyrase is a type IIA topoisomerase and is able to produce negative DNA supercoiling by making a double-strand break in the DNA and religating it. The gyrase enzyme consists of two subunits: the C-terminal GyrA domain that wraps around the DNA strand and the N-terminal GyrB domain that catalyses the ATP-dependant supercoiling of the DNA. CcdB stabilizes the gyrase cleavage complex by binding to the GyrA domain and thus blocks the catalytic function of the gyrase. That means that the gyrase remains bound to the DNA and the cleaved DNA is not religated. DNA- and RNA polymerases can’t copy the DNA anymore and cell proliferation as well as protein biosynthesis is stopped. The double-stranded breaks in the DNA initiate cell death.<p>Because gyrases are specific to bacteria such as E. coli it is also a target for some anti-bacterial medications e.g. ciprofloxacin (CFX). As can be seen in the data below, CcdB proves to be as effective as CFX at inducing DNA cleavage <sup><a href=#681>68.1</a></sup><sup> <a href=#682>68.2</a></sup></p><div class='content-image'align='center'><img src='https://static.igem.org/mediawiki/2013/2/2f/Sspb_CFX_compared.jpg' width='300'>Comparison of the effect of CcdB and CFX on gyrase activity. N: negatively supercoiled DNA, L: linear DNA, SC: supercoiled DNA. A higher concentration of CcdB/CFX leads to more cleaved (linear) DNA<sup><a href= http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1635281/figure/fig2/>[source]</a></sup></div><div class='content-image' align='center'><img src='https://static.igem.org/mediawiki/2013/e/e4/Bonn_Ccdb_and_ccda.jpg' width='300'>  A higher concentration of ccdB leads to blocking of gyrase and positively supercoiled DNA is cleaved to linear DNA instead of being processed to negatively supercoiled DNA. <sup><a href=http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3460896/figure/pone-0046499-g003/>[source]</a></sup></div><p>We used this system to build a light-induced kill-switch. Therefore we added the ssrA tag to the antitoxin ccdA. When the bacteria a emitted to light, ccdA is degraded and ccdB is set free and can bind to the gyrase. and cell death is initiated.  Like in most TA systems, the toxin ccdB is relatively stable, while the antitoxin ccdA is vulnerable to degradation.</p><h2>References</h2><p><a name=681>68.1</a> <a href='http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1635281/'>A strand-passage conformation of DNA gyrase is required to allow the bacterial toxin, CcdB, to access its binding site, Andrew B. Smith and Anthony Maxwell, Nucleic Acids Res. 2006 October; 34(17): 4667–4676, PMC 1635281</a></p><p><a name=682>68.2</a> <a href='http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3460896/'> A Common Origin for the Bacterial Toxin-Antitoxin Systems parD and ccd, Suggested by Analyses of Toxin/Target and Toxin/Antitoxin Interactions, Andew B. Smith et al, PLoS One. 2012; 7(9): e46499, PMCID: PMC3460896</a></p>";
 content.type="Project";
@@ Line 691: / Line 617: @@
 break;
-case 70:
-content.i =70;
-content.parents=[52];
-content.childs=[];
-content.titleShort = "Timeline";
-content.titleLong = "Timeline";
-content.summary= "";
-content.text="";
-content.type="Project";
-break;
 case 71:
@@ Line 753: / Line 669: @@
 content.i = 100;
 content.parents=[37];
-content.childs=[106,107,108,109,101,110,130,112];
+content.childs=[106,107,108,112,101,110,130];
 content.titleShort = "Human Practice";
 content.titleLong = "Human Practice in General";
-content.summary= "The major goal in Human Practice is to make synthetic biology easily understandable and interesting for everybody. For that purpose our project's design follows the well-known franchise of Star Wars; our project name &quot;LOV WARS&quot; was born. The constant references to the Star Wars movies facilitate the access to synthetic biology to a person that is not familiar with the subject. With the most famous icon of Star Wars being the laser sword it offers a great opportunity to present our light induction system in an entertaining way. We put great effort into adapting every aspect of our public presence to Star Wars.";
+content.summary= "For our Human Practice advances our project's design follows the well-known franchise of Star Wars - being inspired by the LOV domain our project name &quot;LOV WARS&quot; was born. The constant references to the Star Wars movies facilitate the access to synthetic biology to a person that is not familiar with the subject. We put great effort into adapting every aspect of our public presence to Star Wars.";
 content.text= "<p>The major goal in Human Practice is to make synthetic biology easily understandable and interesting for everybody. For that purpose our project's design follows the well-known franchise of Star Wars; our project name 'LOV WARS' was born. The constant references to the Star Wars movies facilitate the access to synthetic biology to a person that is not familiar with the subject. With the most famous icon of Star Wars being the laser sword it offers a great opportunity to present our light induction system in an entertaining way. We put great effort into adapting every aspect of our public presence to Star Wars.</p><p>Therefore we introduced a <a onclick=node(110)>flash game</a> and a <a onclick=node(109)>comic series</a> called LOV Wars in which we explain basic concepts of synthetic biology and our project. The comic and the LOV Wars shooter arouse interest in people that would normally never engage with the field. That way we can spread the possibilities and advantages that modern genetically engineered organisms offer and clear possible misunderstandings or prejudices against the subject.</p>For the same purpose we organized multiple events for general public at which we presented the tools used in synthetic biology and our project; over the last six months we <a onclick=node(108)>visited several High Schools</a> in Bonn and nearby and gave presentations. In addition we had an information booth in our city center informing passersby about the risks and benefits of synthetic biology with the aid of several posters and flyers. A big highlight was a <a onclick=node(106)>Science slam</a> we arranged. In front of a big audience six scientists from different fields presented an interesting aspect of their academic field. The Science Slam was a great success in terms of getting people in touch with science and without exception received very good critiques.<br> At every opportunity we handed out questionnaires in which we evaluated the public's opinion about synthetic biology and whether our human practice events were informative and interesting.";
 content.type="Human Practice";
@@ Line 765: / Line 681: @@
 content.parents=[100];
 content.childs=[102,103];
-content.titleShort = "Project presentations"
+content.titleShort = "Presentations"
 content.titleLong = "Project presentations";
 content.summary= "We presented iGEM and our project on two scientific conferences.";
@@ Line 787: / Line 703: @@
 content.parents=[101];
 content.childs=[];
-content.titleShort = "Biocom congress Berlin";
+content.titleShort = "Congress Berlin";
 content.titleLong = "Biocom congress Berlin";
 content.summary= "our day in Berlin. We met important people and other iGEM teams";
@@ Line 812: / Line 728: @@
 content.titleLong = "The Science Slam - Science for Everybody";
 content.summary = "We organized a science slam to offer an experience of science for everyone. And the event met with great approval!";
-content.text = "<h1>Science Slam</h1><p>Our project aims to make versatile control of biological machines easily possible for everybody. During conception we wondered how our human practice work could represent that thought and decided to organize a science slam  an event which makes science available to everybody!</p><p>We believe that science must not only stay in laboratories and lecture halls but reach lots of people out there. And without a doubt, the walls of the reputed ivory tower couldnt stand throughout the evening of our event!</p><p>Furthermore, the science slam provided a platform for our slammers to share the passion for their project with more than just their colleagues and thus help to popularize their field. In ten minutes, they presented their topic in a playful and inspirational manner and the audience decided which slammer has managed to get them carried away the most.</p><p>We managed not only to give floor to a large variety of fields by our slammers from physics, medicine, psychology, informatics, politics and engineering  but to make them be heard by a public which was very mixed indeed.</p><p>As a prelude to the event, we acquainted the audience with synthetic biology and our project by showing what synthetic biology can do for each of us. Distributed questionnaires during the evening showed that -as hoped- our presentation helped to leave a positive mark on the field of synthetic biology in peopleŽs heads:</p><p>For example, 97% of the people, who stated they did not know about synthetic biology before have got a proper picture of it during our event. 86% said, they are now familiar with the iGEM competition and everybody stated, they found the idea to combine the presentation of our iGEM project with a science slam very good or good.</p><p>The fact that our audience liked this topic had been reflected in our voting: Our representative for synthetic biology, Mogan Ramesh won the science slam. He originally comes from engineering but discovered his passion for superbacteria e.coli during his studies which he gladly shared with us. In ten minutes, he taught us that e.coli does not only work as eeew on our meal, but showed how his group studies on making e.coli work for us, e.g. producing pharmaceuticals.</p><p>How? For example, recombinant DNA technology can be used to modify Escherichia coli to produce human insulin. Mogans works within a group of scientist that investigate the metabolic network in the Escherichia coli bacteria cell. Attempts to raise the copy number of plasmid DNA have been successful, and the goal now is to understand how the cell has accomplished this task. Proteomic studies have indicated an expected cellular adaptation that agrees with attempts to raise copy number in the literature; however, the thermodynamic implications of these results have been minimally explored. The goal of a cell remains controversial; one explanation is that the cellular metabolism seeks to maximize the entropy (minimize the Gibbs energy) of the system. Using GAMS as an optimization tool, it is explored if the reaction paths that have proven to be the most likely are also the paths that minimize the Gibbs energy of the system.</p><p>All in all, our science slam has been a great success which demanded a reenactment from a lot of sites.</p><p><b> Here are a few impressions from our event:</b></p><p> <img src=https://static.igem.org/mediawiki/2013/0/07/BonnScienceslam1.PNG> <br /><br /><i>Tobias Bald from experimental dermatology (University of Bonn) spoke about the Dark side of the immune system.</i></p><p><img src=https://static.igem.org/mediawiki/2013/d/dd/BonnScienceslam2.PNG><br /><br /><i>Prof. Dr. Herbert Dreiner gave us a review of Fukushima with full activity.</i></p><p><img src=https://static.igem.org/mediawiki/2013/0/0c/BonnScienceslam3.PNG><br /><br /><i>Our winner Mogan Ramesh shared his passion for superbacteria e.coli.</i></p><p><img src=https://static.igem.org/mediawiki/2013/2/24/BonnScienceslam4.PNG>< br /><br /><i>Full house at our Science Slam.</i></p><p><img src=https://static.igem.org/mediawiki/2013/d/d5/BonnScienceslam5.PNG><br /><br /><i>Our audience obviously enjoyed the show.</i></p>";
+content.text = "<h1>Science Slam</h1><p>Our project aims to make versatile control of biological machines easily possible for everybody. During conception we wondered how our human practice work could represent that thought and decided to organize a science slam  an event which makes science available to everybody!</p><p>We believe that science must not only stay in laboratories and lecture halls but reach lots of people out there. And without a doubt, the walls of the reputed ivory tower couldnt stand throughout the evening of our event!</p><p>Furthermore, the science slam provided a platform for our slammers to share the passion for their project with more than just their colleagues and thus help to popularize their field. In ten minutes, they presented their topic in a playful and inspirational manner and the audience decided which slammer has managed to get them carried away the most.</p><p>We managed not only to give floor to a large variety of fields by our slammers from physics, medicine, psychology, informatics, politics and engineering  but to make them be heard by a public which was very mixed indeed.</p><p>As a prelude to the event, we acquainted the audience with synthetic biology and our project by showing what synthetic biology can do for each of us. Distributed questionnaires during the evening showed that -as hoped- our presentation helped to leave a positive mark on the field of synthetic biology in people&apos;s heads:</p><p>For example, 97% of the people, who stated they did not know about synthetic biology before have got a proper picture of it during our event. 86% said, they are now familiar with the iGEM competition and everybody stated, they found the idea to combine the presentation of our iGEM project with a science slam very good or good. </br> <p> <img src=https://static.igem.org/mediawiki/2013/d/df/Bonn_Science_Slam_1.png> </br> <p> <img src= https://static.igem.org/mediawiki/2013/f/f1/Bonn_Science_Slam_2.png> </br> <p> <img src=https://static.igem.org/mediawiki/2013/5/55/Bonn_Science_Slam_3.png></br> </p><p>The fact that our audience liked this topic had been reflected in our voting: Our representative for synthetic biology, Mogan Ramesh won the science slam. He originally comes from engineering but discovered his passion for superbacteria e.coli during his studies which he gladly shared with us. In ten minutes, he taught us that e.coli does not only work as eeew on our meal, but showed how his group studies on making e.coli work for us, e.g. producing pharmaceuticals.</p><p>How? For example, recombinant DNA technology can be used to modify Escherichia coli to produce human insulin. Mogans works within a group of scientist that investigate the metabolic network in the Escherichia coli bacteria cell. Attempts to raise the copy number of plasmid DNA have been successful, and the goal now is to understand how the cell has accomplished this task. Proteomic studies have indicated an expected cellular adaptation that agrees with attempts to raise copy number in the literature; however, the thermodynamic implications of these results have been minimally explored. The goal of a cell remains controversial; one explanation is that the cellular metabolism seeks to maximize the entropy (minimize the Gibbs energy) of the system. Using GAMS as an optimization tool, it is explored if the reaction paths that have proven to be the most likely are also the paths that minimize the Gibbs energy of the system.</p><p>All in all, our science slam has been a great success which demanded a reenactment from a lot of sites.</p><p><b> Here are a few impressions from our event:</b></p><p> <img src=https://static.igem.org/mediawiki/2013/0/07/BonnScienceslam1.PNG> <br /><br /><i>Tobias Bald from experimental dermatology (University of Bonn) spoke about the Dark side of the immune system.</i></p><p><img src=https://static.igem.org/mediawiki/2013/d/dd/BonnScienceslam2.PNG><br /><br /><i>Prof. Dr. Herbert Dreiner gave us a review of Fukushima with full activity.</i></p><p><img src=https://static.igem.org/mediawiki/2013/0/0c/BonnScienceslam3.PNG><br /><br /><i>Our winner Mogan Ramesh shared his passion for superbacteria e.coli.</i></p><p><img src=https://static.igem.org/mediawiki/2013/2/24/BonnScienceslam4.PNG><br/><br/><i>Full house at our Science Slam.</i></p><p><img src=https://static.igem.org/mediawiki/2013/d/d5/BonnScienceslam5.PNG><br/><br/><i>Our audience obviously enjoyed the show.</i></p>";
 content.type = "Human Practice";
 break;
@@ Line 819: / Line 735: @@
 case 107:
 content.i = 107;
-content.parents=[105];
+content.parents=[100];
+content.childs=[];
 content.titleShort = "SynBio Day";
 content.titleLong = "SynBio Day";
@@ Line 830: / Line 747: @@
 content.parents=[100];
 content.childs=[];
-content.titleShort = "School presentations";
+content.titleShort = "Schools";
 content.titleLong = "School presentations";
 content.summary= "We gave lectures about synthetic biology and our project in schools in order to reach the younger people. ";
@@ Line 840: / Line 757: @@
 case 109:
 content.i = 109;
-content.parents=[105];
+content.parents=[112];
-content.childs=[112];
+content.childs=[];
-content.titleShort = "About the comic";
+content.titleShort = "Comic: About";
 content.titleLong = "About the comic";
 content.summary= "We introduced a hand drawn comic series called &quot;LOV Wars&quot; about Obi-Wan E. Coli and his adventures in the Galaxy of Petri. Basic concepts of synthetic biology are explained along the way.";
-content.text="On the mission to find new and interesting means to bring across the concepts of synthetic biology, we introduced a comic series consisting of three episodes in the style of the well known Star Wars movies. The readers will find themselves in a world where Galaxies are petri dishes and all the characters are bacteria. Alongside the action-filled story we step by step introduce basic concepts of synthetic biology. The use of light sabers and laser guns also offered a great opportunity to embed our system of light-degradable proteins in the plot. At our presentations at schools and at our information booth it proved to be an ideal eye-catcher for passers-by and led to them wanting to know more about the subject.
+content.text="On the mission to find new and interesting means to bring across the concepts of synthetic biology, we introduced a comic series consisting of three episodes in the style of the well known Star Wars movies. The readers will find themselves in a world where Galaxies are petri dishes and all the characters are bacteria. Alongside the action-filled story we step by step introduce basic concepts of synthetic biology. The use of light sabers and laser guns also offered a great opportunity to embed our system of light-degradable proteins in the plot. At our presentations at schools and at our information booth it proved to be an ideal eye-catcher for passers-by and led to them wanting to know more about the subject. During the episodes the reader accompanies the hero Obi-Wan E. Coli and his Padawan Plasmida on their journey through the Galaxy of Petri. They are fighting the villain Darth Cherry and his companions, the clones. But we don&apos;t want to spoil the story for you, just read the comic yourself below."; content.type="Human Practice";
-During the episodes the reader accompanies the hero Obi-Wan E. Coli and his Padawan Plasmida on their journey through the Galaxy of Petri. They are fighting the villain Darth Cherry and his companions, the clones. But we don’t want to spoil the story for you, just read the comic yourself below.";
-content.type="Human Practice";
 break;
@@ Line 854: / Line 769: @@
 content.parents=[100];
 content.childs=[];
-content.titleShort = "About the LOV-Wars Shooter";
+content.titleShort = "LOV-Wars Minigame</br>About";
 content.titleLong = "About the LOV-Wars Shooter";
 content.summary= "The thoughts and ideas behind our game.";
@@ Line 875: / Line 790: @@
 case 112:
 content.i = 112;
-content.parents = [100, 109];
+content.parents = [100];
-content.childs = [];
+content.childs = [109];
 content.titleShort = "Comic";
 content.titleLong = "Comic – The adventures of Obi Wan E.Coli";
 content.summary = "Read about the adventures of Obi Wan E.Coli";
-content.text = "<img src=https://static.igem.org/mediawiki/2013/c/c1/Bonn_Comic_episode1_page1.jpg width=896px><img src=https://static.igem.org/mediawiki/2013/a/ad/Bonn_Comic_episode1_page2.jpg  width=896px><img src=https://static.igem.org/mediawiki/2013/1/19/Bonn_Comic_episode1_page3.jpg width=896px><img src=https://static.igem.org/mediawiki/2013/8/80/Bonn_Comic_episode1_page4.jpg  width=896px><img src=https://static.igem.org/mediawiki/2013/6/6c/Comic_EN_2.0_%281%29.jpg width=896px><img src=https://static.igem.org/mediawiki/2013/8/84/Comic_EN_2.0_%282%29.jpg  width=896px><img src=https://static.igem.org/mediawiki/2013/9/97/Comic_EN_2.0_%283%29.jpg width=896px><img src=https://static.igem.org/mediawiki/2013/1/19/Comic_EN_2.0_%284%29.jpg  width=896px><img src=https://static.igem.org/mediawiki/2013/8/8d/Bonn_Comic_EN_3.0_%281%29.jpg width=896px><img src=https://static.igem.org/mediawiki/2013/5/58/Bonn_Comic_EN_3.0_%282%29.jpg  width=896px><img src=https://static.igem.org/mediawiki/2013/8/8c/Bonn_Comic_EN_3.0_%283%29.jpg width=896px><img src=https://static.igem.org/mediawiki/2013/b/b4/Bonn_Comic_EN_3.0_%284%29.jpg  width=896px>";
+content.text = "<img src=https://static.igem.org/mediawiki/2013/c/c1/Bonn_Comic_episode1_page1.jpg width=735px><img src=https://static.igem.org/mediawiki/2013/a/ad/Bonn_Comic_episode1_page2.jpg  width=735px><img src=https://static.igem.org/mediawiki/2013/1/19/Bonn_Comic_episode1_page3.jpg width=735px><img src=https://static.igem.org/mediawiki/2013/8/80/Bonn_Comic_episode1_page4.jpg  width=735px><img src=https://static.igem.org/mediawiki/2013/6/6c/Comic_EN_2.0_%281%29.jpg width=735px><img src=https://static.igem.org/mediawiki/2013/8/84/Comic_EN_2.0_%282%29.jpg  width=735px><img src=https://static.igem.org/mediawiki/2013/9/97/Comic_EN_2.0_%283%29.jpg width=735px><img src=https://static.igem.org/mediawiki/2013/1/19/Comic_EN_2.0_%284%29.jpg  width=735px><img src=https://static.igem.org/mediawiki/2013/8/8d/Bonn_Comic_EN_3.0_%281%29.jpg width=735px><img src=https://static.igem.org/mediawiki/2013/5/58/Bonn_Comic_EN_3.0_%282%29.jpg  width=735px><img src=https://static.igem.org/mediawiki/2013/8/8c/Bonn_Comic_EN_3.0_%283%29.jpg width=735px><img src=https://static.igem.org/mediawiki/2013/b/b4/Bonn_Comic_EN_3.0_%284%29.jpg  width=735px>";
 content.type = "Human Practice";
 break;
@@ Line 889: / Line 804: @@
 content.i = 130;
 content.parents=[100];
-content.childs=[131,133,134,135,136,137,138];
+content.childs=[131,133,136,137,138];
 content.titleShort = "About us";
 content.titleLong = "About us";
 content.summary= "The Team - Presentation of all team members";
-content.text= "<div><div style='float:right;display:inline; '><img src='https://static.igem.org/mediawiki/2013/a/a4/Bonn_sponsors_uniBonn.png' class='bottom-sponsor' height='50px' id='sponsor-unibonn' style='margin-right:10px;'><img src='https://static.igem.org/mediawiki/2013/7/79/Bonn_sponsors_limes.gif' class='bottom-sponsor' height='50px' id='sponsor-limes'></div><div class='subpage-head-link'><a href='javascript:history.back();'>Back</a></br> Category: Team</div></html>= About Us =<html><div class='subpage-text'><div align='center'><div width:870px; height:485px; id='aboutus-group' style='position:relative'><img src='https://static.igem.org/mediawiki/2013/e/e9/Teammod.jpg' width='870px' style='z-index:1;position:relative;'><div id='aboutus-group-kristina' onmouseover=showMemberDetails('Kristina') onmouseout=hideMemberDetails() style='position:absolute;top:160px;left:57px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-dustin' onmouseover=showMemberDetails('Dustin') onmouseout=hideMemberDetails() style='position:absolute;top:153px;left:130px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-marc' onmouseover=showMemberDetails('Marc') onmouseout=hideMemberDetails() style='position:absolute;top:275px;left:124px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-niklas' onmouseover=showMemberDetails('Niklas') onmouseout=hideMemberDetails() style='position:absolute;top:139px;left:174px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-ben' onmouseover=showMemberDetails('Ben') onmouseout=hideMemberDetails() style='position:absolute;top:158px;left:197px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-annika' onmouseover=showMemberDetails('Annika') onmouseout=hideMemberDetails() style='position:absolute;top:278px;left:220px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-matthias' onmouseover=showMemberDetails('Matthias') onmouseout=hideMemberDetails() style='position:absolute;top:153px;left:252px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-max' onmouseover=showMemberDetails('Max') onmouseout=hideMemberDetails() style='position:absolute;top:159px;left:282px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-maria' onmouseover=showMemberDetails('Maria') onmouseout=hideMemberDetails() style='position:absolute;top:273px;left:289px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-sadrija' onmouseover=showMemberDetails('Sadrija') onmouseout=hideMemberDetails() style='position:absolute;top:160px;left:358px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-franzi' onmouseover=showMemberDetails('Franzi') onmouseout=hideMemberDetails() style='position:absolute;top:277px;left:355px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-cori' onmouseover=showMemberDetails('Cori') onmouseout=hideMemberDetails() style='position:absolute;top:172px;left:420px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-pauline' onmouseover=showMemberDetails('Pauline') onmouseout=hideMemberDetails() style='position:absolute;top:282px;left:421px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-olli' onmouseover=showMemberDetails('Olli') onmouseout=hideMemberDetails() style='position:absolute;top:163px;left:472px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-nina' onmouseover=showMemberDetails('Nina') onmouseout=hideMemberDetails() style='position:absolute;top:277px;left:489px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-miriam' onmouseover=showMemberDetails('Miriam') onmouseout=hideMemberDetails() style='position:absolute;top:163px;left:506px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-thomas' onmouseover=showMemberDetails('Thomas') onmouseout=hideMemberDetails() style='position:absolute;top:140px;left:535px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-katharina' onmouseover=showMemberDetails('Katharina') onmouseout=hideMemberDetails() style='position:absolute;top:276px;left:557px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-philipp' onmouseover=showMemberDetails('Philipp') onmouseout=hideMemberDetails() style='position:absolute;top:147px;left:591px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-martina' onmouseover=showMemberDetails('MartinaB') onmouseout=hideMemberDetails() style='position:absolute;top:162px;left:622px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-sebastian' onmouseover=showMemberDetails('Sebastian') onmouseout=hideMemberDetails() style='position:absolute;top:153px;left:664px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-annikag' onmouseover=showMemberDetails('AnnikaG') onmouseout=hideMemberDetails() style='position:absolute;top:150px;left:703px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-jan' onmouseover=showMemberDetails('Jan') onmouseout=hideMemberDetails() style='position:absolute;top:136px;left:769px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-details' style='background-color:black;opacity:0.8;position:absolute;height:80px;width=870px;top:0px;left:0px;right:-24px;z-index:2;'><div id='aboutus-group-details-name' style='color:white'></div><div id='aboutus-group-details-course' style='color:white'></div><div id='aboutus-group-details-semester' style='color:white'></div><div id='aboutus-group-details-tasks' style='color:white'></div></div><div id='aboutus-group-move' style='background-color:black;opacity:0.8;position:absolute;height:20px;width=870px;top:0px;left:0px;right:-24px;z-index:2;'><div id='aboutus-group-about' style='color:white'>Hover over faces to show group member details.</div></div></div></div></div><div id='team-members'><div class='team-members-row'><div class='team-member'><div class='team-member-name'>Thomas Berger</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork, jamboree presentation, school presentations</div></div><div class='team-member'><div class='team-member-name'>Sadrija Cukoski</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>4th semester</div><div class='job'>science slam</div></div><div class='team-member'><div class='team-member-name'>Dustin Dankelmann</div><div class='team-member-field'>Bachelor Biology</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork, financing</div></div></div><div class='team-members-row'><div class='team-member'><div class='team-member-name'>Katharina Düker</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>4th semester</div><div class='job'>startup advisory</div></div><div class='team-member'><div class='team-member-name'>Maria Gädeke</div><div class='team-member-field'>Bachelor Biology</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork</div></div><div class='team-member'><div class='team-member-name'>Jan Hansen</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork, webdesign, lov-wars shooter, design, comic, school presentations, pictures</div></div></div><div class='team-members-row'><div class='team-member'><div class='team-member-name'>Kristina Klein</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork</div></div><div class='team-member'><div class='team-member-name'>Matthias Klumpp</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>4th semester</div><div class='job'>webmaster</div></div><div class='team-member'><div class='team-member-name'>Franziska Kohl</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>4th semester</div><div class='job'>survey and evaluation, school presentations</div></div></div><div class='team-members-row'><div class='team-member'><div class='team-member-name'>Sebastian Martin</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>4th semester</div><div class='job'>safety</div></div><div class='team-member'><div class='team-member-name'>Miriam Melake</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>2nd semester</div><div class='job'>science slam</div></div><div class='team-member'><div class='team-member-name'>Nina Offermann</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork, comic, school presentations</div></div></div><div class='team-members-row'><div class='team-member'><div class='team-member-name'>Oliver Rippel</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork, school presentations</div></div><div class='team-member'><div class='team-member-name'>Philipp Sander</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>4th semester</div><div class='job'>startup advisory</div></div><div class='team-member'><div class='team-member-name'>Florian Schäfer</div><div class='team-member-field'>Bachelor Mathematics</div><div class='team-member-sem'>6th semester</div><div class='job'>modelling</div></div></div><div class='team-members-row'><div class='team-member'><div class='team-member-name'>Max Schelski</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>4th semester</div><div class='job'>management, labwork, jamboree presentation, webdesign, school presentations</div></div><div class='team-member'><div class='team-member-name'>Niklas Schmacke</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>4th semester</div><div class='job'>labwork, jamboree presentation, webdesign, design</div></div><div class='team-member'><div class='team-member-name'>Corinna Schmalohr</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork, comic, school presentation, design</div></div></div><div class='team-members-row'><div class='team-member'><div class='team-member-name'>Annika Schneider</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork, jamboree presentation, meetings, financing</div></div><div class='team-member'><div class='team-member-name'>Marc Schulte</div><div class='team-member-field'>Master Molecular Biotechnology</div><div class='team-member-sem'>4th semester</div><div class='job'>labwork, design</div></div><div class='team-member'><div class='team-member-name'>Benjamin Syllwasschy</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork, school presentations</div></div></div><div class='team-members-row'><div class='team-member'><div class='team-member-name'>Pauline Ulmke</div><div class='team-member-field'>Bachelor Applied Biology</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork, school presentation</div></div><div class='team-member'><div class='team-member-name'>Leonie von Berlin</div><div class='team-member-field'>Bachelor Biology</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork</div></div></div></div></div></div></div></div>";
+content.text= "<div><div class='subpage-text'><div align='center'><div width:870px; height:485px; id='aboutus-group' style='position:relative;left:-90px'><img src='https://static.igem.org/mediawiki/2013/e/e9/Teammod.jpg' width='870px' style='z-index:1;position:relative;'><div id='aboutus-group-kristina' onmouseover=showMemberDetails('Kristina') onmouseout=hideMemberDetails() style='position:absolute;top:160px;left:57px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-dustin' onmouseover=showMemberDetails('Dustin') onmouseout=hideMemberDetails() style='position:absolute;top:153px;left:130px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-marc' onmouseover=showMemberDetails('Marc') onmouseout=hideMemberDetails() style='position:absolute;top:275px;left:124px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-niklas' onmouseover=showMemberDetails('Niklas') onmouseout=hideMemberDetails() style='position:absolute;top:139px;left:174px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-ben' onmouseover=showMemberDetails('Ben') onmouseout=hideMemberDetails() style='position:absolute;top:158px;left:197px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-annika' onmouseover=showMemberDetails('Annika') onmouseout=hideMemberDetails() style='position:absolute;top:278px;left:220px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-matthias' onmouseover=showMemberDetails('Matthias') onmouseout=hideMemberDetails() style='position:absolute;top:153px;left:252px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-max' onmouseover=showMemberDetails('Max') onmouseout=hideMemberDetails() style='position:absolute;top:159px;left:282px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-maria' onmouseover=showMemberDetails('Maria') onmouseout=hideMemberDetails() style='position:absolute;top:273px;left:289px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-sadrija' onmouseover=showMemberDetails('Sadrija') onmouseout=hideMemberDetails() style='position:absolute;top:160px;left:358px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-franzi' onmouseover=showMemberDetails('Franzi') onmouseout=hideMemberDetails() style='position:absolute;top:277px;left:355px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-cori' onmouseover=showMemberDetails('Cori') onmouseout=hideMemberDetails() style='position:absolute;top:172px;left:420px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-pauline' onmouseover=showMemberDetails('Pauline') onmouseout=hideMemberDetails() style='position:absolute;top:282px;left:421px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-olli' onmouseover=showMemberDetails('Olli') onmouseout=hideMemberDetails() style='position:absolute;top:163px;left:472px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-nina' onmouseover=showMemberDetails('Nina') onmouseout=hideMemberDetails() style='position:absolute;top:277px;left:489px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-miriam' onmouseover=showMemberDetails('Miriam') onmouseout=hideMemberDetails() style='position:absolute;top:163px;left:506px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-thomas' onmouseover=showMemberDetails('Thomas') onmouseout=hideMemberDetails() style='position:absolute;top:140px;left:535px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-katharina' onmouseover=showMemberDetails('Katharina') onmouseout=hideMemberDetails() style='position:absolute;top:276px;left:557px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-philipp' onmouseover=showMemberDetails('Philipp') onmouseout=hideMemberDetails() style='position:absolute;top:147px;left:591px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-martina' onmouseover=showMemberDetails('MartinaB') onmouseout=hideMemberDetails() style='position:absolute;top:162px;left:622px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-sebastian' onmouseover=showMemberDetails('Sebastian') onmouseout=hideMemberDetails() style='position:absolute;top:153px;left:664px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-annikag' onmouseover=showMemberDetails('AnnikaG') onmouseout=hideMemberDetails() style='position:absolute;top:150px;left:703px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-jan' onmouseover=showMemberDetails('Jan') onmouseout=hideMemberDetails() style='position:absolute;top:136px;left:769px;width:30px;height:30px;z-index:5;'></div><div id='aboutus-group-details' style='background-color:black;opacity:0.8;position:absolute;height:80px;width=870px;top:0px;left:0px;right:-24px;z-index:2;'><div id='aboutus-group-details-name' style='color:white'></div><div id='aboutus-group-details-course' style='color:white'></div><div id='aboutus-group-details-semester' style='color:white'></div><div id='aboutus-group-details-tasks' style='color:white'></div></div><div id='aboutus-group-move' style='background-color:black;opacity:0.8;position:absolute;height:20px;width=870px;top:0px;left:0px;right:-24px;z-index:2;'><div id='aboutus-group-about' style='color:white'>Hover over faces to show group member details.</div></div></div></div></div><div id='team-members'><div class='team-members-row'><div class='team-member'><div class='team-member-name'>Thomas Berger</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork, jamboree presentation, school presentations</div></div><div class='team-member'><div class='team-member-name'>Sadrija Cukoski</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>4th semester</div><div class='job'>science slam</div></div><div class='team-member'><div class='team-member-name'>Dustin Dankelmann</div><div class='team-member-field'>Bachelor Biology</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork, financing</div></div></div><div class='team-members-row'><div class='team-member'><div class='team-member-name'>Katharina Düker</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>4th semester</div><div class='job'>startup advice, skills course, school presentation</div></div><div class='team-member'><div class='team-member-name'>Maria Gädeke</div><div class='team-member-field'>Bachelor Biology</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork</div></div><div class='team-member'><div class='team-member-name'>Jan Hansen</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork, webdesign, lov-wars shooter, design, comic, school presentations, pictures</div></div></div><div class='team-members-row'><div class='team-member'><div class='team-member-name'>Kristina Klein</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork</div></div><div class='team-member'><div class='team-member-name'>Matthias Klumpp</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>4th semester</div><div class='job'>webmaster</div></div><div class='team-member'><div class='team-member-name'>Franziska Kohl</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>4th semester</div><div class='job'>survey and evaluation, school presentations</div></div></div><div class='team-members-row'><div class='team-member'><div class='team-member-name'>Sebastian Martin</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>4th semester</div><div class='job'>safety</div></div><div class='team-member'><div class='team-member-name'>Miriam Melake</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>2nd semester</div><div class='job'>science slam</div></div><div class='team-member'><div class='team-member-name'>Nina Offermann</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork, comic, school presentations</div></div></div><div class='team-members-row'><div class='team-member'><div class='team-member-name'>Oliver Rippel</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork, school presentations</div></div><div class='team-member'><div class='team-member-name'>Philipp Sander</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>4th semester</div><div class='job'>startup advice, financing, skills course, school presentation</div></div><div class='team-member'><div class='team-member-name'>Florian Schäfer</div><div class='team-member-field'>Bachelor Mathematics</div><div class='team-member-sem'>6th semester</div><div class='job'>modelling</div></div></div><div class='team-members-row'><div class='team-member'><div class='team-member-name'>Max Schelski</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>4th semester</div><div class='job'>management, labwork, jamboree presentation, jamboree poster, webdesign, school presentations, skills course</div></div><div class='team-member'><div class='team-member-name'>Niklas Schmacke</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>4th semester</div><div class='job'>labwork, jamboree presentation, jamboree poster webdesign, design</div></div><div class='team-member'><div class='team-member-name'>Corinna Schmalohr</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork, comic, school presentation, design</div></div></div><div class='team-members-row'><div class='team-member'><div class='team-member-name'>Annika Schneider</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork, jamboree presentation, meetings, financing</div></div><div class='team-member'><div class='team-member-name'>Marc Schulte</div><div class='team-member-field'>Master Molecular Biotechnology</div><div class='team-member-sem'>4th semester</div><div class='job'>labwork, design</div></div><div class='team-member'><div class='team-member-name'>Benjamin Syllwasschy</div><div class='team-member-field'>Bachelor Molecular Biomedicine</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork, school presentations</div></div></div><div class='team-members-row'><div class='team-member'><div class='team-member-name'>Pauline Ulmke</div><div class='team-member-field'>Bachelor Applied Biology</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork, school presentation</div></div><div class='team-member'><div class='team-member-name'>Leonie von Berlin</div><div class='team-member-field'>Bachelor Biology</div><div class='team-member-sem'>2nd semester</div><div class='job'>labwork</div></div></div></div></div></div>";
 content.type="Team";
 break;
@@ Line 915: / Line 830: @@
 content.titleLong = "Sponsors";
 content.summary= "Many sponsors made our work possible.";
-content.text= "<div class=subpage-text> <table class=subpage-sponsors>  <tr class=subpage-sponsor style=border-style:solid;border-width:5px;border-color:grey;> <td class=subpage-sponsor> <img src=https://static.igem.org/mediawiki/2013/3/36/Bonn_sponsor_promega.png class=bottom-sponsor width=300px id=sponsor-limes> </td> <td> <h2>Promega</h2> </br> Promega is one of the five biggest worldwide acting Life Science Research company. It was founded in Madison, WI (USA) and produces products and system solutions for gen-, protein- and cell-analysis. With the help of these products biological systems can be explored easily. Promega-products can be used in basic research, development of medicaments, molecular diagnostic and identification of human genetic constitution. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/3/36/Bonn_sponsors_eppendorf.jpg class=bottom-sponsor width=300px id=sponsor-eppendorf> </td> <td> <h2>Eppendorf</h2> </br> Eppendorf is a biotechnical company that develops, produces and sells systems for life science research for laboratories all over the world. The assortment of goods contains pipettes, dispensers, centrifuges, reaction tubes and pipette tips. Moreover Eppendorf offers instruments and systems to manipulate cells, automated machines for Liquid Handling and for DNA- Amplification, as well as Biochips. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/5/5a/Bonn_sponsor_IKA.png class=bottom-sponsor width=300px id=sponsor-ika> </td> <td><h2> IKA </h2> </br> In 1910 the companys history of IKA began, China had not been invented yet Peoples Republic and the word globalization. Today, the IKA group about 800 employees at eight locations on four continents and is pleased with clients such as BASF, Bayer and Procter & Gamble. In most product groups, we are sovereign world market leader and a symbol of development and growth. Or as our new slogan: &quot;IKA - Designed to work perfectly.&quot; </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/b/bf/Bonn_sponsors_Ella.jpg class=bottom-sponsor width=300px'id=sponsor-ella> </td> <td> <h2>Ella Biotech</h2> </br> ELLA is an independent, privately owned company founded in October 2004. ELLA offers creative services for the production of oligonucleotides driven by the goal of continually improving our production strategies. ELLA offers tangible advantages to its customers and partners through its validated technology platform, its experienced interdisciplinary team, and its resolute attitude towards the highest quality in our products. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/c/c0/Bonn_sponsor_genscript.jpg class=bottom-sponsor width=370px id=sponsor-genscript> </td> <td> <h2> Gen Script </h2> </br> GenScript is a leading biology CRO focusing exclusively on early drug discovery and development services. Built on our assembly-line mode, one-stop solution, continuous improvement, and stringent IP protection, GenScript provides a comprehensive portfolio of services that include Bio-Reagent, Bio-Assay, Lead Optimization, and Antibody Drug Development which can be effectively integrated into your value chain and your operations. We strive with competence and confidence to meet your demand for developing pre-clinical drug candidates time-efficiently and cost-effectively. With track performance record, GenScript is your ideal and reliable innovation partner in drug discovery. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/1/13/Bonn_sponsor_idt.jpg class=bottom-sponsor width=300px id=sponsor-idt> </td> <td><h2> IDT </h2> </br> Genetically engineered vaccines and pharmaceuticals for the global marketplace. The IDT Biologicals is an innovative medium-sized company that is involved in more than 90 years with its products and services to the health maintenance of humans and animals. Genetically engineered vaccines and pharmaceuticals for the domestic and international market are manufactured. In fiscal year 2012, the IDT biologics had a turnover of around 151 million euros. In the IDT biologics around 1,100 people are currently employed. IDT Biologics is an independent company of the Klocke Group, which developed and implemented at five production sites innovative packaging solutions for the pharmaceutical, cosmetic, food and chemical-technical industry.</br> Decades of research and development of vaccines</br> Since the beginning of the IDT successfully fought the various pathogens in animals with complex vaccine development from the laboratory to production and sales in one location. From research and development to manufacturing and testing and approval, national and international marketing of the range of tasks. The IDT Animal Health operates its own modern research complex for the development of animal vaccines.</br> Integrated biopharmaceutical services. </br> Since its founding in 1921, the IDT has biologics developed into a center for the pharmaceutical and biotechnology with the divisions Animal Health, human vaccines and pharmaceuticals. More than 250 million euros have been invested since privatization in 1993 in the continuous expansion of an integrated biopharmaceutical site and thus not only created excellent conditions of production, but also highly modern workplaces. </td>  </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/3/3a/Bonn_sponsor_neb.jpg class=bottom-sponsor width=300px id=sponsor-neb> </td> <td> <h2> New England Biolabs </h2> </br> Never before in the history of science, the demands on the molecular biology industry have been as high as today: The researcher asks for the absolute best and most reliable products - there is no room for compromise. New England Biolabs fulfills this requirement. For 35 years we have been a leader in the development and production of enzymes for molecular biology and other reagents in the &quot;life sciences&quot; such as for proteomics and drug discovery. Our expertise in enzyme technology based on our strategic cloning and expression of DNA-Restriktions-/Modifikationssystemen program. So we have for years set the standards in terms of quality and price. </td>  </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/3/34/Bonn_sponsors_MN.png class=bottom-sponsor width=300px id=sponsor-mn> </td> <td> <h2> Macherey und Nagel </h2> </br> MACHEREY-NAGEL is a family-run concern in the fourth generation. The comprehensive portfolio includes the areas of filtration, rapid tests, water analysis, chromatography and bioanalysis. MACHEREY-NAGEL employs more than 470 highly skilled employees in sales, production as well as research and development, including 10% post-doctoral researchers. They all guarantee an exceptional service. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/3/37/Bonn_Sponsor_roche.jpg class=bottom-sponsor width=300px id=sponsor-roche> </td> <td> <h2> Roche </h2> </br> Roche Headquartered in Basel, Switzerland, is a leading research-focused healthcare company with the pharmaceuticals and diagnostics businesses. As the worlds largest biotech company developing clinically differentiated medicines in oncology, virology, inflammation, metabolism and central nervous system. Roche, a pioneer in diabetes management, is also the world leader in in-vitro diagnostics, tissue-based cancer diagnostics. Medicines and diagnostic tools that enable tangible improvements in the health, quality of life and survival of patients is the strategic goal of personalized medicine from Roche. This concept is based on new molecular insights and molecular diagnostic tests that allow a more precise tuning of therapy and better control of the disease. Therapies are tailored to patient groups that have similarities in their disease. The only way to improve the efficacy of drugs targeted and maintain quality of life. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/d/d2/Bonn_sponsor_geneious.gif class=bottom-sponsor width=300px id=sponsor-geneious>  </td> <td> <h2> Geneious </h2> </br> In Good Company</br> First released in 2005, Geneious is one the worlds leading bioinformatics software platforms, used by over 2500 universities and institutes and commercial companies in more than 65 countries. Geneious is used by all 20 of the top 20 Universities globally (Times Higher Education, 2012) and by seven of the 10 largest pharmaceutical companies.</br> Dedicated to excellence</br> Our software has won a number of prestigious awards, including the Computerworld Excellence Awards from Innovative Use of ICT and the United Nations World Summer Awards and Winner in the e-Science Category in 2007, the Recruit IT Innovative Software Product Award at the PriceWaterhouseCoopers Hi-Tech Awards in 2009 and a Global Finalist in the IT and Informatics category at the Bio-IT World Awards in Boston in 2009.  </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/5/55/Bonn_Sponsor_vwr.jpg class=bottom-sponsor width=300px id=sponsor-vwr> </td> <td> <h2> VWR </h2> </br> VWR is science for the advancement of the worlds most important research through the distribution of a wide range of products and services to a variety of important companies in the pharmaceutical, biotechnology, and healthcare industries as well as government agencies, universities and schools. We offer our customers all the resources they need to be successful, ie an extensive range of the best products in the areas of chemicals, furniture, appliances, instruments, apparel and consumables, from a wide variety of leaders in the field of science manufacturers. With 160 years of experience in this industry, VWR further supports its customers through a combination of strength, vision, innovation and a well-established distribution network that reaches thousands of specialized labs and facilities across the planet. VWR is not just a product supplier - it keeps the most important research in the world in motion. VWRs expertise in the areas of supply chain and logistics services enables customers to fully concentrate on their areas of expertise. Of the management of procurement processes to the integration of supply chains: VWR helps specialized research facilities and laboratories to work with maximum efficiency. VWR has over 8,000 employees in 30 countries with direct offices throughout the world working to streamline the way, as researchers from North America, Europe and the Asia-Pacific region supply and maintain their labs. In addition, VWR further supports its customers by providing onsite services, storeroom management, product procurement, supply chain systems integration and technical services. We are expanding our global presence and adhere to the principle that customers benefit from the availability and expertise of our local sales teams.</br> In todays economy VWR helps its customers to focus on increasing productivity and reducing costs and optimizing procurement processes. </br> Headquartered in Radnor, PA (USA), earned VWR International, LLC, in 2012 global sales of more than 4.1 billion U.S. dollars. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/0/0b/Bonn_Sponsors_Roth.png class=bottom-sponsor width=170px id=sponsor-roth> </td> <td> <h2> ROTH – A COMPANY WITH TRADITION </h2> </br> 1879 </br> Carl ROTH founded in Karlsruhe, a &quot;material, Colonial and dye business and Droguerie&quot;.</br>1899</br> The first sales and mail order catalog is published.</br> 1956</br> Publication of the first issue of the publication a &quot;Rarea &quot; Natural Products</br> 1990</br> First ROTH general catalog. Complete with the areas of laboratory, life sciences and chemicals in one.</br> 2005</br> Completion of a modern establishment for the production and storage of laboratory chemicals and reagents in Karlsruhe Rhine port area. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/7/79/Bonn_starlab.jpg class=bottom-sponsor width=300px id=sponsor-starlab> </td> <td> <h2> Starlab </h2> </br> STARLAB is a company specializing in liquid handling technology group. With subsidiaries in Germany, France, Britain and Italy is available in the direct sales an extensive range of products available. Plus, you get our products to many countries around the world via our international trading partners. Our success is based on many years of experience in manufacturing and marketing of liquid handling disposable products - with TipOne we have established ourselves as a leading supplier of pipette tips systems worldwide. This quality, price and service come first. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/b/bf/Bonn_caesar.gif class=bottom-sponsor width=200px id=sponsor-casar> </td> <td> <h2> Caesar </h2> </br> The center of advanced european studies and research (caesar) is an institute of the Max Planck Society, which is located at the boundaries between neuroscience, cell biology and biophysics. The focus of the research is the particular cellular and neural signal processing. </br> Caesar works with modern photonic, molecular biological, chemical and micro-technological methods. The focus of kinetic, spectroscopic and microscopic methods are research and control of cellular activity. </td>  </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/6/6e/Bonn_Eurofins.png class=bottom-sponsor width=300px id=sponsor-eurofins> </td> <td> <h2> Eurofins MWG operon </h2> </br> Discover our fascination about the world of the four bases.</br> We are fascinated about the power of DNA and how it is incorporated in everything we do, work and live. Being passionate about our strong customer orientation, our service and our quality standards, we continuously challenge ourselves to stay ahead and remain one of the leading genomics service providers worldwide. Eurofins MWG Operon is globally known for its innovative and customised technologies in the life science industries and academic research institutions. With the combined power of an international network of Eurofins companies in the field of genomic services, forensics, agroscience, pharmaceutical, environmental, food and feed testing, we have established an outstanding team of experts and broad range of technologies. This unique constellation underlines our approach to offer best practise solutions and versatile concepts for our clients - academic institutions and large interdisciplinary operating companies of the world. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/1/18/Bonn_ThermoFisher.jpg class=bottom-sponsor width=300px id=sponsor-thermofisher> </td>  <td> <h2> Thermo Fisher </h2> </br> Thermo Fisher Scientific Inc. (NYSE: TMO) is the world leader in serving science. Our mission is to enable our customers to make the world healthier, cleaner and safer. With revenues of $13 billion, we have approximately 39,000 employees and serve customers within pharmaceutical and biotech companies, hospitals and clinical diagnostic labs, universities, research institutions and government agencies, as well as in environmental and process control industries. We create value for our key stakeholders through three premier brands, Thermo Scientific, Fisher Scientific and Unity<sup> TM </sup> Lab Services, which offer a unique combination of innovative technologies, convenient purchasing options and a single solution for laboratory operations management. Our products and services help our customers solve complex analytical challenges, improve patient diagnostics and increase laboratory productivity. </td></tr></table> </div> </div>  </div> </div> </div> ";
+content.text= "<div class=subpage-text> <table class=subpage-sponsors style=margin-left:-50px;background-color:transparent><tr><td colspan=2><h2>Our Partners</h2></td></tr><tr class=subpage-sponsor style=border-style:solid;border-width:5px;border-color:grey;> <td colspan=2 class=subpage-sponsor>   <h2>BIO.NRW Cluster Biotechnology North Rhine-Westphalia</h2> <img src=https://static.igem.org/mediawiki/2013/9/90/Bonn_sponsor_BioNRW.png class=bottom-sponsor width=500px id=sponsor-limes style=margin-bottom:-50px></td><tr><td colspan=2></br>North Rhine-Westphalia is situated at Europe’s geographic and economic center. It is the largest of Germany’s 16 federal states, and the number one exporter. If classified as an independent exporting nation, NRW would rank 18th in the world, on a par with the Netherlands.</br>North Rhine-Westphalia’s state government has established a number of technology clusters to systematically improve NRW’s strengths and talents in established industries and up-and-coming fields like biotechnology. Goal of the “ExcellenceNRW” cluster strategy is to create a favourable climate for innovation, as that is the best way to sustain the competitive edge and stimulate growth and employment in the companies that call the state their home.North Rhine-Westphalia’s biotechnology cluster BIO.NRW is a central catalyst for the sustainable development of the state’s biotech sector. It activates cooperation between business, research, investors and policy-makers. The cluster also promotes the strengths and achievements of biotechnology in the state.</br>To support young as well as already established biotech companies, BIO.NRW offers the following core competencies:</br>– Individual matchmaking for collaborations and partners</br>– Overview on all up-to-date R&D activities in industry and academia</br>– Profound knowledge on financing possibilities</br>– International promotion and marketing for NRW as biotech location</br>– Direct contacts to decision makers</br><h3>Our services include</h3><bold>Technology Transfer</bold></br>Tech transfer support is a key contribution from BIO.NRW. We organize events, working platforms and meetings to promote the dialogue between all stakeholders in the field of biotechnology and to encourage cooperation.</br><bold>Biotech Business & Sciences</bold></br>BIO.NRW compiles comprehensive and current online databases of the academic institutions and companies active in the life sciences in NRW. Free to access and easy-to-use, these resources are valuable tools for identifying prospective business partners. More information on <a href=http://www.bio.nrw.de>www.bio.nrw.de</a></br><bold>Fairs, Exhibitions and Conferences</bold></br>Companies and academic institutions can generate awareness of their activities locally, nationally and internationally by being a part of the BIO.NRW common stands on fairs, exhibitions and conferences. BIO.NRW also organizes a series of workshops and symposia, called BIO.NRW.academy.</br><bold>Support of Young Professionals</bold></br>BIO.NRW takes a special interest in supporting young professionals in biotechnology. For example, we organize conventions where graduates meet representatives from industry and academic science. The ‘Business Angel Network – BIO.NRW’ helps financing and funding biotech start-ups. In addition, a forum that brings together investment institutions, private investors and business angels and developers provides information about the current NRW biotech scene. These meetings are a valuable opportunity for start-up companies to receive coaching and financing.</br></br>To learn more about the Cluster BIO.NRW and to stay informed about the latest biotech developments in NRW please visit <a href=http://www.bio.nrw.de>www.bio.nrw.de</a>.</td> </tr><tr><td colspan=2><h2>Our Sponsors</h2></td></tr><tr class=subpage-sponsor style=border-style:solid;border-width:5px;border-color:grey;> <td class=subpage-sponsor> <img src=https://static.igem.org/mediawiki/2013/3/36/Bonn_sponsor_promega.png class=bottom-sponsor width=250px id=sponsor-limes> </td> <td> <h2>Promega</h2> </br> Promega is one of the five biggest worldwide acting Life Science Research company. It was founded in Madison, WI (USA) and produces products and system solutions for gen-, protein- and cell-analysis. With the help of these products biological systems can be explored easily. Promega-products can be used in basic research, development of medicaments, molecular diagnostic and identification of human genetic constitution. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/3/36/Bonn_sponsors_eppendorf.jpg class=bottom-sponsor width=250px id=sponsor-eppendorf> </td> <td> <h2>Eppendorf</h2> </br> Eppendorf is a biotechnical company that develops, produces and sells systems for life science research for laboratories all over the world. The assortment of goods contains pipettes, dispensers, centrifuges, reaction tubes and pipette tips. Moreover Eppendorf offers instruments and systems to manipulate cells, automated machines for Liquid Handling and for DNA- Amplification, as well as Biochips. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/5/5a/Bonn_sponsor_IKA.png class=bottom-sponsor width=250px id=sponsor-ika> </td> <td><h2> IKA </h2> </br> In 1910 the companys history of IKA began, China had not been invented yet Peoples Republic and the word globalization. Today, the IKA group about 800 employees at eight locations on four continents and is pleased with clients such as BASF, Bayer and Procter & Gamble. In most product groups, we are sovereign world market leader and a symbol of development and growth. Or as our new slogan: &quot;IKA - Designed to work perfectly.&quot; </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/b/bf/Bonn_sponsors_Ella.jpg class=bottom-sponsor width=250px'id=sponsor-ella> </td> <td> <h2>Ella Biotech</h2> </br> ELLA is an independent, privately owned company founded in October 2004. ELLA offers creative services for the production of oligonucleotides driven by the goal of continually improving our production strategies. ELLA offers tangible advantages to its customers and partners through its validated technology platform, its experienced interdisciplinary team, and its resolute attitude towards the highest quality in our products. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/c/c0/Bonn_sponsor_genscript.jpg class=bottom-sponsor width=370px id=sponsor-genscript> </td> <td> <h2> Gen Script </h2> </br> GenScript is a leading biology CRO focusing exclusively on early drug discovery and development services. Built on our assembly-line mode, one-stop solution, continuous improvement, and stringent IP protection, GenScript provides a comprehensive portfolio of services that include Bio-Reagent, Bio-Assay, Lead Optimization, and Antibody Drug Development which can be effectively integrated into your value chain and your operations. We strive with competence and confidence to meet your demand for developing pre-clinical drug candidates time-efficiently and cost-effectively. With track performance record, GenScript is your ideal and reliable innovation partner in drug discovery. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/1/13/Bonn_sponsor_idt.jpg class=bottom-sponsor width=250px id=sponsor-idt> </td> <td><h2> IDT </h2> </br> Genetically engineered vaccines and pharmaceuticals for the global marketplace. The IDT Biologicals is an innovative medium-sized company that is involved in more than 90 years with its products and services to the health maintenance of humans and animals. Genetically engineered vaccines and pharmaceuticals for the domestic and international market are manufactured. In fiscal year 2012, the IDT biologics had a turnover of around 151 million euros. In the IDT biologics around 1,100 people are currently employed. IDT Biologics is an independent company of the Klocke Group, which developed and implemented at five production sites innovative packaging solutions for the pharmaceutical, cosmetic, food and chemical-technical industry.</br> Decades of research and development of vaccines</br> Since the beginning of the IDT successfully fought the various pathogens in animals with complex vaccine development from the laboratory to production and sales in one location. From research and development to manufacturing and testing and approval, national and international marketing of the range of tasks. The IDT Animal Health operates its own modern research complex for the development of animal vaccines.</br> Integrated biopharmaceutical services. </br> Since its founding in 1921, the IDT has biologics developed into a center for the pharmaceutical and biotechnology with the divisions Animal Health, human vaccines and pharmaceuticals. More than 250 million euros have been invested since privatization in 1993 in the continuous expansion of an integrated biopharmaceutical site and thus not only created excellent conditions of production, but also highly modern workplaces. </td>  </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/3/3a/Bonn_sponsor_neb.jpg class=bottom-sponsor width=250px id=sponsor-neb> </td> <td> <h2> New England Biolabs </h2> </br> Never before in the history of science, the demands on the molecular biology industry have been as high as today: The researcher asks for the absolute best and most reliable products - there is no room for compromise. New England Biolabs fulfills this requirement. For 35 years we have been a leader in the development and production of enzymes for molecular biology and other reagents in the &quot;life sciences&quot; such as for proteomics and drug discovery. Our expertise in enzyme technology based on our strategic cloning and expression of DNA-Restriktions-/Modifikationssystemen program. So we have for years set the standards in terms of quality and price. </td>  </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/3/34/Bonn_sponsors_MN.png class=bottom-sponsor width=250px id=sponsor-mn> </td> <td> <h2> Macherey und Nagel </h2> </br> MACHEREY-NAGEL is a family-run concern in the fourth generation. The comprehensive portfolio includes the areas of filtration, rapid tests, water analysis, chromatography and bioanalysis. MACHEREY-NAGEL employs more than 470 highly skilled employees in sales, production as well as research and development, including 10% post-doctoral researchers. They all guarantee an exceptional service. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/3/37/Bonn_Sponsor_roche.jpg class=bottom-sponsor width=250px id=sponsor-roche> </td> <td> <h2> Roche </h2> </br> Roche Headquartered in Basel, Switzerland, is a leading research-focused healthcare company with the pharmaceuticals and diagnostics businesses. As the worlds largest biotech company developing clinically differentiated medicines in oncology, virology, inflammation, metabolism and central nervous system. Roche, a pioneer in diabetes management, is also the world leader in in-vitro diagnostics, tissue-based cancer diagnostics. Medicines and diagnostic tools that enable tangible improvements in the health, quality of life and survival of patients is the strategic goal of personalized medicine from Roche. This concept is based on new molecular insights and molecular diagnostic tests that allow a more precise tuning of therapy and better control of the disease. Therapies are tailored to patient groups that have similarities in their disease. The only way to improve the efficacy of drugs targeted and maintain quality of life. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/d/d2/Bonn_sponsor_geneious.gif class=bottom-sponsor width=250px id=sponsor-geneious>  </td> <td> <h2> Geneious </h2> </br> In Good Company</br> First released in 2005, Geneious is one the worlds leading bioinformatics software platforms, used by over 2500 universities and institutes and commercial companies in more than 65 countries. Geneious is used by all 20 of the top 20 Universities globally (Times Higher Education, 2012) and by seven of the 10 largest pharmaceutical companies.</br> Dedicated to excellence</br> Our software has won a number of prestigious awards, including the Computerworld Excellence Awards from Innovative Use of ICT and the United Nations World Summer Awards and Winner in the e-Science Category in 2007, the Recruit IT Innovative Software Product Award at the PriceWaterhouseCoopers Hi-Tech Awards in 2009 and a Global Finalist in the IT and Informatics category at the Bio-IT World Awards in Boston in 2009.  </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/5/55/Bonn_Sponsor_vwr.jpg class=bottom-sponsor width=250px id=sponsor-vwr> </td> <td> <h2> VWR </h2> </br> VWR is science for the advancement of the worlds most important research through the distribution of a wide range of products and services to a variety of important companies in the pharmaceutical, biotechnology, and healthcare industries as well as government agencies, universities and schools. We offer our customers all the resources they need to be successful, ie an extensive range of the best products in the areas of chemicals, furniture, appliances, instruments, apparel and consumables, from a wide variety of leaders in the field of science manufacturers. With 160 years of experience in this industry, VWR further supports its customers through a combination of strength, vision, innovation and a well-established distribution network that reaches thousands of specialized labs and facilities across the planet. VWR is not just a product supplier - it keeps the most important research in the world in motion. VWRs expertise in the areas of supply chain and logistics services enables customers to fully concentrate on their areas of expertise. Of the management of procurement processes to the integration of supply chains: VWR helps specialized research facilities and laboratories to work with maximum efficiency. VWR has over 8,000 employees in 30 countries with direct offices throughout the world working to streamline the way, as researchers from North America, Europe and the Asia-Pacific region supply and maintain their labs. In addition, VWR further supports its customers by providing onsite services, storeroom management, product procurement, supply chain systems integration and technical services. We are expanding our global presence and adhere to the principle that customers benefit from the availability and expertise of our local sales teams.</br> In todays economy VWR helps its customers to focus on increasing productivity and reducing costs and optimizing procurement processes. </br> Headquartered in Radnor, PA (USA), earned VWR International, LLC, in 2012 global sales of more than 4.1 billion U.S. dollars. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/0/0b/Bonn_Sponsors_Roth.png class=bottom-sponsor width=170px id=sponsor-roth> </td> <td> <h2> ROTH – A COMPANY WITH TRADITION </h2> </br> 1879 </br> Carl ROTH founded in Karlsruhe, a &quot;material, Colonial and dye business and Droguerie&quot;.</br>1899</br> The first sales and mail order catalog is published.</br> 1956</br> Publication of the first issue of the publication a &quot;Rarea &quot; Natural Products</br> 1990</br> First ROTH general catalog. Complete with the areas of laboratory, life sciences and chemicals in one.</br> 2005</br> Completion of a modern establishment for the production and storage of laboratory chemicals and reagents in Karlsruhe Rhine port area. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/7/79/Bonn_starlab.jpg class=bottom-sponsor width=250px id=sponsor-starlab> </td> <td> <h2> Starlab </h2> </br> STARLAB is a company specializing in liquid handling technology group. With subsidiaries in Germany, France, Britain and Italy is available in the direct sales an extensive range of products available. Plus, you get our products to many countries around the world via our international trading partners. Our success is based on many years of experience in manufacturing and marketing of liquid handling disposable products - with TipOne we have established ourselves as a leading supplier of pipette tips systems worldwide. This quality, price and service come first. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/b/bf/Bonn_caesar.gif class=bottom-sponsor width=200px id=sponsor-casar> </td> <td> <h2> Caesar </h2> </br> The center of advanced european studies and research (caesar) is an institute of the Max Planck Society, which is located at the boundaries between neuroscience, cell biology and biophysics. The focus of the research is the particular cellular and neural signal processing. </br> Caesar works with modern photonic, molecular biological, chemical and micro-technological methods. The focus of kinetic, spectroscopic and microscopic methods are research and control of cellular activity. </td>  </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/6/6e/Bonn_Eurofins.png class=bottom-sponsor width=250px id=sponsor-eurofins> </td> <td> <h2> Eurofins MWG operon </h2> </br> Discover our fascination about the world of the four bases.</br> We are fascinated about the power of DNA and how it is incorporated in everything we do, work and live. Being passionate about our strong customer orientation, our service and our quality standards, we continuously challenge ourselves to stay ahead and remain one of the leading genomics service providers worldwide. Eurofins MWG Operon is globally known for its innovative and customised technologies in the life science industries and academic research institutions. With the combined power of an international network of Eurofins companies in the field of genomic services, forensics, agroscience, pharmaceutical, environmental, food and feed testing, we have established an outstanding team of experts and broad range of technologies. This unique constellation underlines our approach to offer best practise solutions and versatile concepts for our clients - academic institutions and large interdisciplinary operating companies of the world. </td> </tr> <tr class=subpage-sponsor> <td> <img src=https://static.igem.org/mediawiki/2013/1/18/Bonn_ThermoFisher.jpg class=bottom-sponsor width=250px id=sponsor-thermofisher> </td>  <td> <h2> Thermo Fisher </h2> </br> Thermo Fisher Scientific Inc. (NYSE: TMO) is the world leader in serving science. Our mission is to enable our customers to make the world healthier, cleaner and safer. With revenues of $13 billion, we have approximately 39,000 employees and serve customers within pharmaceutical and biotech companies, hospitals and clinical diagnostic labs, universities, research institutions and government agencies, as well as in environmental and process control industries. We create value for our key stakeholders through three premier brands, Thermo Scientific, Fisher Scientific and Unity<sup> TM </sup> Lab Services, which offer a unique combination of innovative technologies, convenient purchasing options and a single solution for laboratory operations management. Our products and services help our customers solve complex analytical challenges, improve patient diagnostics and increase laboratory productivity. </td></tr></table> </div> </div>  </div> </div> </div> ";
 content.type="Team";
 break;
@@ Line 936: / Line 851: @@
 content.childs=[];
 content.titleShort = "Bonn";
-content.titleLong = "Bonn";
+content.titleLong = "Our city Bonn";
-content.summary= "Description of Bonn";
+content.summary= "Bonn is a beautiful city in North Rhine-Westphalia, Germany.";
-content.text= "Founded by the Romans in the year 12 before Christ, birthplace of Beethoven, once Capitol of Germany- now: Bonn is a vivid place to life and study. Next to Cologne, Bonn is also set on the river Rhine. Everybody gets smitten with it´s charm- such a unique mixture of tradition and modern lifestyle.</br>Stadt. City. Ville. Bonn.";
+content.text= "Bonn has a long and eventful history. The city was founded by the Romans in the year 12 before Christ and continued to grow ever since. Bonn also was the birthplace of the famous musician and composer Beethoven and even was the Capitol of West Germany for 41 years. Today Bonn has around 300 000 habitants and has become the cultural of the region. It is set on the river Rhine very close to Cologne. Everybody gets smitten with its charm and unique mixture of tradition and modern lifestyle. The city&apos;s historic center and the setting close to the river Rhine turn it into a wonderful place to live and study.<p style='text-align:center;'>the city&apos;s slogan:</br><i>&quot;Stadt. City. Ville. Bonn&quot;</i></p></br><p><a href=https://static.igem.org/mediawiki/2013/0/0b/Bonn_by_night.JPG><div class='content-image'align='center'><img src='https://static.igem.org/mediawiki/2013/0/0b/Bonn_by_night.JPG' width='600'></a>Bonn by night <sup>&copy;Jan Hansen</sup></div></p><p><div class='content-image'align='center'><a href=https://static.igem.org/mediawiki/2013/a/a6/Bonn_poppelsdorfer_schloss.JPG><img src='https://static.igem.org/mediawiki/2013/a/a6/Bonn_poppelsdorfer_schloss.JPG' width='600'></a>The university <sup>&copy;Jan Hansen</sup></div><p><a href=https://static.igem.org/mediawiki/2013/0/0b/Bonn_by_night.JPG><div class='content-image'align='center'><img src='https://static.igem.org/mediawiki/2013/7/75/Bonn_beethoven.JPG' width='600'></a>The famous Beethoven statue in the city center <sup><a href=http://www.bone-diagenesis.org/moziloCMS/gallery.php?gal=Bonn-nbsp~Impressions>[source]</a></sup></div></p><h2>References</h2><p><a name='1'>1.</a> <a href='http://www.bonn.de'>http://www.bonn.de</a></p><p><a name='2'>2.</a> <a href='http://de.wikipedia.org/wiki/Bonn'>http://de.wikipedia.org/wiki/Bonn</a></p>";
 content.type="Team";
 break;
@@ Line 990: / Line 905: @@
 break; */
+/*
 case 112:
 content.i = 112;
-content.parents = [100, 109];
+content.parents = [100];
-content.childs = [];
+content.childs = [109];
 content.titleShort = "Comic";
 content.titleLong = "Comic – The adventures of Obi Wan E.Coli";
@@ Line 1,001: / Line 916: @@
 content.type = "Human Practice";
 break;
+*/
+case 200:
+content.i = 200;
+content.parents=[37];
+content.childs=[];
+content.titleShort = "Safety";
+content.titleLong = "Safety";
+content.summary= "Since safety is an important concern for any iGEM team, we gave deep thought to it. In this article our reflections about safety measures are presented.";
+content.text= " Safety is an important issue for all of the IGEM teams. Taking care of the health and safety of our teammates is of the utmost importance for us. In our opinion, only a safe lab is a good environment for research and work. On our safety page we would like to give you a brief overview over the precautions we take to ensure a safe work environment in the lab. </br></br> <h2> A </h2> Do the biological materials used in your lab work pose any of the following risks: </br> </br> 1. Risks to the safety and health of team members or others working in the lab? </br> In our lab, we work with E. coli (strain: DH5alpha;XL1Blue;w3110 sspB-; w3110 sspB- LacI-DAS+4;Top10; DH10 Beta). All these E.Coli strains are classified as risk group 1, as they can cause irritation of the skin, the eyes, and the respiratory tract; additionally, they can affect the kidneys. Although our E.coli strains are nonpathogenic, we wear protective clothes and examination gloves to ensure safety. </br> We also use ethidum bromide in our lab, which is toxic and can cause cancer. To minimize the risk of accidental contact with this dangerous substance, we have fume hoods especially designated for working with ethidium bromide. Furthermore, the use of nitrile gloves as well as lab coats is mandatory when we are working with potentially harmful substances and any waste contaminated with ethidium bromide is kept separate from other waste. </br> In addition, every team member had to take a course about biosafety and regulations in Germany (as required in §8-12 of the GenTSV, the German act of genetic engineering). </br></br> 2. Risks to the safety and health of the general public, if released by design or by accident? </br> There is little risk to the general public, since our bacteria strains are nonpathogenic and in general do not cause harm to humans. And yet, the contact of an immunocompromised person with our bacteria strains could lead to health issues. </br></br> 3. Risk to the environment, if released by design or by accident? </br> Since our bacteria and genetic parts are non pathogenic and the light induced protein degradation is not a threat to the environment, there is minimal risk to the environment if our (modified) bacteria were released . In addition, our lab is equipped with a special air conditioning system which maintains a slight low pressure environment in the lab and thus prevents the air from inside of the lab from getting out when the doors are opened. The risk of (genetically altered) bacteria being released by accident is therefore reduced. </br> </br> 4.Risks to security through malicious misuse by individuals, groups, or countries? </br> The risk of malicious misuse can never be completely eliminated. Although there are cameras around the campus as well as other security installations, an element of risk remains. If our project was stolen and our bacteria were released, the risks for the public would be minimal, since our bacteria strains are nonpathogenic and our project mainly deals with the light-inducible degradation of proteins, which is hardly dangerous. </br></br> 5. If your project moved from a small-scale lab study to become widely used as a commercial/industrial product, what new risks might arise? </br> Since our light-inducible degradation system is mostly thought to be applicable in restricted areas (labs, bioreactors) the risks are low. Of course, any industrial application of our system should mot be designed without additional fail-safes to ensure multiple layers of safety. </br> One possible “risk” of our project is inherent to any other biological system; namely, that mutations in our device's gene sequence lead to unforeseen consequences. We assume that it is most likely that a mutation in our device will lead to the device's failure, i.e. leading to an accumulation of the protein that is supposed to be degraded. Whether this poses merely efficiency problems or more serious biosafety risks depends on the specific application and should be considered by persons responsible for implementing that instance of our technology. </br></br> 6. Does your project include any design features to address safety risks? (For example: kill switches, auxotrophic chassis, etc.) Note that including such features is not mandatory to participate in iGEM, but many groups choose to include them.  </br> We developed a kill switch that is induced by blue light. This switch is capable of killing bacteria that are accidentally removed from a restricted area - for example a red-light hood – and which are then exposed to daylight in a short period of time. </br></br> 7. What safety training have you received (or plan to receive in the future)? Provide a brief description, and a link to your institution’s safety training requirements, if available. </br> Every team member had to take a course about safe work in the lab. We also have advisors who teach us how to work properly and safely and inform us about safety measures, such as wearing protective clothing. We also plan to implement a disinfection checkpoint that our team members will have to pass when leaving our entering the lab to decrease the chance of contamination. </br></br> 8. Under what biosafety provisions will / do you work? </br> Please provide a link to your institution biosafety guidelines. </br> The institution's biosafety guidelines are the national safety guidelines. <sup><a href=#2001>200.1</a></sup> </br> </br></br> <h2>B</h2> Does your institution have an Institutional Biosafety Committee, or an equivalent group? If yes, have you discussed your project with them? Describe any concerns they raised with your project, and any changes you made to your project plan based on their review. </br> Our university does not have a committee for biosafety. We are using the guidelines from the central biosafety committee from our federal government. We also work together with our supervisor, discussing our project and to address any safety issues that might arise. </br></br></br> <h2>C</h2> Does your country have national biosafety regulations or guidelines? If so, please provide a link to these regulations or guidelines if possible. <sup><a href=#2001>200.1</a></sup> <sup><a href=#2002>200.2</a></sup> </br></br></br> <h2>D</h2> According to the WHO Biosafety Manual, what is the BioSafety Level rating of your lab? </br> Our Biosafety Level is 1. <h2><b>References</b></h2></br> <a name=2001>200.1</a> <a href= http://www.gesetze-im-internet.de/gentsv/index.html#BJNR023400990BJNE001503320>  http://www.gesetze-im-internet.de/gentsv/index.html#BJNR023400990BJNE001503320 </a> </br> <a name=2002>200.2</a> <a href= http://www.gesetze-im-internet.de/gentsv/anhang_iii_34.html>  http://www.gesetze-im-internet.de/gentsv/anhang_iii_34.html </a>";
+content.type="Project";
+break;
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Template:Team:Bonn:NetworkData

From 2013.igem.org

Latest revision as of 13:27, 1 December 2013