http://2012.igem.org/wiki/index.php?title=Team:HKUST-Hong_Kong/Design_Chassis&feed=atom&action=historyTeam:HKUST-Hong Kong/Design Chassis - Revision history2024-03-28T10:14:49ZRevision history for this page on the wikiMediaWiki 1.16.0http://2012.igem.org/wiki/index.php?title=Team:HKUST-Hong_Kong/Design_Chassis&diff=211068&oldid=prevDynastywarrior at 19:07, 26 September 20122012-09-26T19:07:37Z<p></p>
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<td colspan='2' style="background-color: white; color:black;">← Older revision</td>
<td colspan='2' style="background-color: white; color:black;">Revision as of 19:07, 26 September 2012</td>
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<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><p><a href="https://2012.igem.org/Team:HKUST-Hong_Kong"><b>Home</b></a></p></ins></div></td></tr>
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<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><li><p><b>Team</b><p><ol></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><li><p><a href="https://2012.igem.org/Team:HKUST-Hong_Kong/Introduction">Introduction</a></p></li></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><li><p><a href="https://2012.igem.org/Team:HKUST-Hong_Kong/Supervisor">Supervisor</a></p></li></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><li><p><a href="https://2012.igem.org/Team:HKUST-Hong_Kong/Instructor">Instructor</a></p></li></ins></div></td></tr>
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<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><li><p><b>Project</b></p><ol></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><li><p><a href="https://2012.igem.org/Team:HKUST-Hong_Kong/Project_Abstraction">Abstract</a></p></li></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><li><p><a href="https://2012.igem.org/Team:HKUST-Hong_Kong/Background_and_Motive">Motive</a></p></li></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><li><p><a href="https://2012.igem.org/Team:HKUST-Hong_Kong/Design_Overview">Design - Overview</a></p></li></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><li><p><a href="https://2012.igem.org/Team:HKUST-Hong_Kong/Design_Module">Design - Module</a></p></li></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><p>-- <a href="https://2012.igem.org/Team:HKUST-Hong_Kong/Module/Target_binding">Target Binding Module</a></p></ins></div></td></tr>
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<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><li><p><a href="https://2012.igem.org/Team:HKUST-Hong_Kong/Notebook">Notebook</a></p></li></ins></div></td></tr>
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</table>Dynastywarriorhttp://2012.igem.org/wiki/index.php?title=Team:HKUST-Hong_Kong/Design_Chassis&diff=208664&oldid=prevHyht2011 at 18:18, 26 September 20122012-09-26T18:18:20Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><br><b>Low degree of virulence.</b></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><br><b>Low degree of virulence.</b></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>As a natural member of the human gut microbiome, <i>B. subtilis</i> is considered to be non-pathogenic to humans. There are very few cases of humans being infected by <i>B. subtilis</i>, and of those who have been infected the vast majority had severe immune deficiency. According to Edberg (1991), <i>B. subtilis</i> does not produce significant quantities of extracellular enzymes or possess other virulence factors that would predispose it to cause infection. In other word, <i>B. subtilis</i> possesses low virulence to humans and has a low risk of adverse effects to human health. <br></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>As a natural member of the human gut microbiome, <i>B. subtilis</i> is considered to be non-pathogenic to humans. There are very few cases of humans being infected by <i>B. subtilis</i>, and of those who have been infected the vast majority had severe immune deficiency. According to Edberg (<ins class="diffchange diffchange-inline">Edberg </ins>1991), <i>B. subtilis</i> does not produce significant quantities of extracellular enzymes or possess other virulence factors that would predispose it to cause infection. In other word, <i>B. subtilis</i> possesses low virulence to humans and has a low risk of adverse effects to human health. <br></div></td></tr>
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</table>Hyht2011http://2012.igem.org/wiki/index.php?title=Team:HKUST-Hong_Kong/Design_Chassis&diff=208584&oldid=prevHyht2011 at 18:16, 26 September 20122012-09-26T18:16:37Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><br><b>References.</b><br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><br><b>References.</b><br></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>Collins M. D. and Gibson G. R.. 1999. Probiotics, prebiotics and synbiotics: Approaches for modulating the microbial ecology of the gut. <i>Am. J. Clin. Nutr.<i> 69:1052S–1057S </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>Collins M. D. and Gibson G. R.. 1999. Probiotics, prebiotics and synbiotics: Approaches for modulating the microbial ecology of the gut. <i>Am. J. Clin. Nutr.<<ins class="diffchange diffchange-inline">/</ins>i> 69:1052S–1057S </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Corazza G.R., Benati G., Strocchi A., Sorge M. & Gasbarrini G.. 1992. Treatment with <i>Bacillus subtilis</i> reduces intestinal hydrogen production in patients with gaseous symptoms. <i>Current therapeutic research</i>. 52.1: 144-151</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Corazza G.R., Benati G., Strocchi A., Sorge M. & Gasbarrini G.. 1992. Treatment with <i>Bacillus subtilis</i> reduces intestinal hydrogen production in patients with gaseous symptoms. <i>Current therapeutic research</i>. 52.1: 144-151</div></td></tr>
</table>Hyht2011http://2012.igem.org/wiki/index.php?title=Team:HKUST-Hong_Kong/Design_Chassis&diff=208193&oldid=prevScarim at 18:08, 26 September 20122012-09-26T18:08:06Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p><i>B. subtilis</i> does not only exist widely in nature, but also contributes to part of the normal flora in the gut (Collin and Gibson 1999). Regarded as a probiotic, it has been proved to have positive effects on patients suffering from functional abdominal bloating (Corazza et al. 1992). Using <i>B. subtilis</i> as our chassis will in theory mean we will not be introducing any exotic species into the gut. <br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p><i>B. subtilis</i> does not only exist widely in nature, but also contributes to part of the normal flora in the gut (Collin and Gibson 1999). Regarded as a probiotic, it has been proved to have positive effects on patients suffering from functional abdominal bloating (Corazza et al. 1992). Using <i>B. subtilis</i> as our chassis will in theory mean we will not be introducing any exotic species into the gut. <br></div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><br><b>References.</<del class="diffchange diffchange-inline">strong</del>><br></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><br><b>References.</<ins class="diffchange diffchange-inline">b</ins>><br></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>Collins M. D. and Gibson G. R.. 1999. Probiotics, prebiotics and synbiotics: Approaches for modulating the microbial ecology of the gut. Am. J. Clin. Nutr. 69:1052S–1057S </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>Collins M. D. and Gibson G. R.. 1999. Probiotics, prebiotics and synbiotics: Approaches for modulating the microbial ecology of the gut. <ins class="diffchange diffchange-inline"><i></ins>Am. J. Clin. Nutr.<ins class="diffchange diffchange-inline"><i> </ins>69:1052S–1057S </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>Corazza G.R., Benati G., Strocchi A., Sorge M. & Gasbarrini G.. 1992. Treatment with Bacillus subtilis reduces intestinal hydrogen production in patients with gaseous symptoms. Current therapeutic research. 52.1: 144-151</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>Corazza G.R., Benati G., Strocchi A., Sorge M. & Gasbarrini G.. 1992. Treatment with <ins class="diffchange diffchange-inline"><i></ins>Bacillus subtilis<ins class="diffchange diffchange-inline"></i> </ins>reduces intestinal hydrogen production in patients with gaseous symptoms. <ins class="diffchange diffchange-inline"><i></ins>Current therapeutic research<ins class="diffchange diffchange-inline"></i></ins>. 52.1: 144-151</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>Edberg, S.C.. 1991. US EPA human health assessment: Bacillus subtilis. Unpublished, U.S. Environmental Protection Agency, Washington, D.C. </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>Edberg, S.C.. 1991. US EPA human health assessment: <ins class="diffchange diffchange-inline"><i></ins>Bacillus subtilis<ins class="diffchange diffchange-inline"></i></ins>. Unpublished, U.S. Environmental Protection Agency, Washington, D.C. </div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>Pooley H. M., Merchante R. and Karamata D.. 1996. Overall protein content and induced enzyme components of the periplasm of Bacillus subtilis. Microb. Drug Resist.2:9–15. </p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>Pooley H. M., Merchante R. and Karamata D.. 1996. Overall protein content and induced enzyme components of the periplasm of <ins class="diffchange diffchange-inline"><i></ins>Bacillus subtilis<ins class="diffchange diffchange-inline"></i></ins>. <ins class="diffchange diffchange-inline"><i></ins>Microb. Drug Resist<ins class="diffchange diffchange-inline"></i></ins>.2:9–15. </p></div></td></tr>
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</table>Scarimhttp://2012.igem.org/wiki/index.php?title=Team:HKUST-Hong_Kong/Design_Chassis&diff=208123&oldid=prevScarim at 18:06, 26 September 20122012-09-26T18:06:24Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>We chose <i>Bacillus subtilis</i> - a model Gram-positive bacterial species often used in laboratory study - as our chassis. It is ideal as a base for designing mechanisms to fulfill our project aims for reasons we mention below.<br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>We chose <i>Bacillus subtilis</i> - a model Gram-positive bacterial species often used in laboratory study - as our chassis. It is ideal as a base for designing mechanisms to fulfill our project aims for reasons we mention below.<br></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><br><b>Safety.</b<del class="diffchange diffchange-inline">></p</del>></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><br><b>Safety.</b></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>The safety issues concerning synthetic biology are numerous, particularly when it comes to the area of medical treatment. To reduce the chance of recombinant bacteria causing harm, synthetic biologists try to develop and utilize solutions that minimize the potential hazards of their products. Regarding safety issues, we the HKUST iGEM 2012 team chose to use <i>B. subtilis</i>, a non-pathogenic chassis.<br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>The safety issues concerning synthetic biology are numerous, particularly when it comes to the area of medical treatment. To reduce the chance of recombinant bacteria causing harm, synthetic biologists try to develop and utilize solutions that minimize the potential hazards of their products. Regarding safety issues, we the HKUST iGEM 2012 team chose to use <i>B. subtilis</i>, a non-pathogenic chassis.<br></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><br>Low degree of virulence.</<del class="diffchange diffchange-inline">p</del>></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><br<ins class="diffchange diffchange-inline">><b</ins>>Low degree of virulence.</<ins class="diffchange diffchange-inline">b</ins>></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>As a natural member of the human gut microbiome, <i>B. subtilis</i> is considered to be non-pathogenic to humans. There are very few cases of humans being infected by <i>B. subtilis</i>, and of those who have been infected the vast majority had severe immune deficiency. According to Edberg (1991), <i>B. subtilis</i> does not produce significant quantities of extracellular enzymes or possess other virulence factors that would predispose it to cause infection. In other word, <i>B. subtilis</i> possesses low virulence to humans and has a low risk of adverse effects to human health. <del class="diffchange diffchange-inline"></p></del><br></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>As a natural member of the human gut microbiome, <i>B. subtilis</i> is considered to be non-pathogenic to humans. There are very few cases of humans being infected by <i>B. subtilis</i>, and of those who have been infected the vast majority had severe immune deficiency. According to Edberg (1991), <i>B. subtilis</i> does not produce significant quantities of extracellular enzymes or possess other virulence factors that would predispose it to cause infection. In other word, <i>B. subtilis</i> possesses low virulence to humans and has a low risk of adverse effects to human health. <br></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><br><<del class="diffchange diffchange-inline">p</del>>Establishment of integration vectors.</p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><br><<ins class="diffchange diffchange-inline">b</ins>>Establishment of integration vectors.</<ins class="diffchange diffchange-inline">b></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><</ins>p<ins class="diffchange diffchange-inline">>The discovery of natural integration in <i>B. subtilis</i> raised great attention in the field of molecular genetics in 1978. A series of integration vectors for <i>B. subtilis</i> were designed later for different purposes. For our project we employed an integration vector not only for its stability in <i>B. subtilis</i>, but also its advantages in safety. The employment of the integration vector to some extent minimizes the risk of antibiotic resistance spreading within the normal flora in gut. In addition, since the exposure of BMP2 to normal tissues can induce adverse effects, integrating target genes into the genome can reduce the chance of spreading the <i>Bmp2</i> gene through horizontal gene transfer and avoid non-specific drug release in gut.<br</ins>></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><<del class="diffchange diffchange-inline">p align="left"</del>><del class="diffchange diffchange-inline">The discovery of natural integration in </del><<del class="diffchange diffchange-inline">em</del>><del class="diffchange diffchange-inline">Bacillus subtilis</del></<del class="diffchange diffchange-inline">em</del>> <del class="diffchange diffchange-inline">raised great attention in the field of molecular genetics in 1978. Due to its natural integration, a series of integration vectors for </del><<del class="diffchange diffchange-inline">em</del>><del class="diffchange diffchange-inline">B</del>. <del class="diffchange diffchange-inline">subtilis</del></<del class="diffchange diffchange-inline">em</del>> <del class="diffchange diffchange-inline">were designed later for different purposes. In our project</del>, <del class="diffchange diffchange-inline">we employed an integration vector not only for its stability </del>in <<del class="diffchange diffchange-inline">em</del>>B. subtilis</<del class="diffchange diffchange-inline">em</del>><del class="diffchange diffchange-inline">, but also </del>its <del class="diffchange diffchange-inline">advantages </del>in <del class="diffchange diffchange-inline">safety. The employment of integration vector </del>to <del class="diffchange diffchange-inline">some extent minimizes </del>the <del class="diffchange diffchange-inline">risk of antibiotic resistance spreading within the normal flora in gut</del>. <del class="diffchange diffchange-inline">In addition</del>, <del class="diffchange diffchange-inline">since the exposure of BMP2 to normal tissues can induce adverse effect, integrating target genes into genome can reduce the chance of spreading of </del><i><del class="diffchange diffchange-inline">Bmp2</del></i> <del class="diffchange diffchange-inline">gene through horizontal gene transfer and avoid non</del>-<del class="diffchange diffchange-inline">specific drug </del> <del class="diffchange diffchange-inline">release </del>in <del class="diffchange diffchange-inline">gut</del>. <<del class="diffchange diffchange-inline">/p></ol</del>></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><<ins class="diffchange diffchange-inline">br</ins>><<ins class="diffchange diffchange-inline">b</ins>><ins class="diffchange diffchange-inline">Protein secretion.</ins></<ins class="diffchange diffchange-inline">b</ins>></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><<ins class="diffchange diffchange-inline">p</ins>><ins class="diffchange diffchange-inline">Compared with <i>E</ins>. <ins class="diffchange diffchange-inline">coli</ins></<ins class="diffchange diffchange-inline">i</ins>>, <ins class="diffchange diffchange-inline">another commonly used chassis </ins>in <ins class="diffchange diffchange-inline">iGEM, </ins><<ins class="diffchange diffchange-inline">i</ins>>B. subtilis</<ins class="diffchange diffchange-inline">i</ins>> <ins class="diffchange diffchange-inline">is preferred because of </ins>its <ins class="diffchange diffchange-inline">reliability </ins>in <ins class="diffchange diffchange-inline">secreting proteins directly </ins>to the <ins class="diffchange diffchange-inline">extracellular environment</ins>. <ins class="diffchange diffchange-inline">As a Gram-positive eubacteria</ins>, <i><ins class="diffchange diffchange-inline">B. subtilis</ins></i> <ins class="diffchange diffchange-inline">lacks an outer membrane, featuring only a 10</ins>-<ins class="diffchange diffchange-inline">50nm peptidoglycan layer beyond its the plasma membrane. </ins> <ins class="diffchange diffchange-inline">Since we are aiming for bacterial secretion of BMP2 out into the digestive tract </ins>in <ins class="diffchange diffchange-inline">the vicinity of colon cancer cells, this property suits our purposes well</ins>.<<ins class="diffchange diffchange-inline">br</ins>></div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"> </del><<del class="diffchange diffchange-inline">li</del>><del class="diffchange diffchange-inline">Protein secretion:</del></<del class="diffchange diffchange-inline">li</del>></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><<ins class="diffchange diffchange-inline">br</ins>><ins class="diffchange diffchange-inline"><b>Peptide display.</ins></<ins class="diffchange diffchange-inline">b></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><p>Without an outer membrane, <i>B. subtilis</i> is ideal for displaying items on its surface. The cell wall of <i>B. subtilis</i> is the surface of the bacterium and contains approximately 9% of the total protein in the cell. The discovery and study of cell wall binding devices produced from cell wall bound proteins provides an attractive tool for surface display of peptide (Pooley et al. 1996). Aiming to display the tumor-binding peptide, RPMrel, on bacteria surface, we decided to take advantage of the cell wall display system, LytC, in <i>B. subtilis</i>. Thus, the ease of peptide display serves as one of our reasons to choose <i>B. subtilis</i> as our chassis. <br</ins>></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><<del class="diffchange diffchange-inline">p</del>><del class="diffchange diffchange-inline">Comparing with </del><<del class="diffchange diffchange-inline">em</del>><del class="diffchange diffchange-inline">E</del>. <del class="diffchange diffchange-inline">coli</del></<del class="diffchange diffchange-inline">em</del>><del class="diffchange diffchange-inline">, another commonly used chassis in iGEM, </del><<del class="diffchange diffchange-inline">em</del>><del class="diffchange diffchange-inline">Bacillus subtilis</del><<del class="diffchange diffchange-inline">/em> is preferred because of its high ability in secreting proteins to extracellular environment. As a Gram-positive eubacteria, <em</del>>B. <del class="diffchange diffchange-inline"> </del>subtilis</<del class="diffchange diffchange-inline">em</del>> <del class="diffchange diffchange-inline">lacks an outer membrane</del>, <del class="diffchange diffchange-inline">only with </del>a <del class="diffchange diffchange-inline">layer of 10 </del>to <del class="diffchange diffchange-inline">50nm peptidoglycan outside cytoplasmic membrane</del>. <del class="diffchange diffchange-inline">Thus </del><<del class="diffchange diffchange-inline">em</del>><del class="diffchange diffchange-inline">Bacillus </del>subtilis</<del class="diffchange diffchange-inline">em</del>> <del class="diffchange diffchange-inline">can secrete proteins directly </del>into the <del class="diffchange diffchange-inline">extracellular environment more easily</del>. <del class="diffchange diffchange-inline">Since we are aiming for bacterial secretion of BMP2 out to the colon, this property suits our purposes well. </del><<del class="diffchange diffchange-inline">/p</del>></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><<ins class="diffchange diffchange-inline">br</ins>><<ins class="diffchange diffchange-inline">b</ins>><ins class="diffchange diffchange-inline">Part of gut commensal flora</ins>.</<ins class="diffchange diffchange-inline">b</ins>></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><<ins class="diffchange diffchange-inline">p</ins>><<ins class="diffchange diffchange-inline">i</ins>>B. subtilis</<ins class="diffchange diffchange-inline">i</ins>> <ins class="diffchange diffchange-inline">does not only exist widely in nature</ins>, <ins class="diffchange diffchange-inline">but also contributes to part of the normal flora in the gut (Collin and Gibson 1999). Regarded as </ins>a <ins class="diffchange diffchange-inline">probiotic, it has been proved </ins>to <ins class="diffchange diffchange-inline">have positive effects on patients suffering from functional abdominal bloating (Corazza et al</ins>. <ins class="diffchange diffchange-inline">1992). Using </ins><<ins class="diffchange diffchange-inline">i</ins>><ins class="diffchange diffchange-inline">B. </ins>subtilis</<ins class="diffchange diffchange-inline">i</ins>> <ins class="diffchange diffchange-inline">as our chassis will in theory mean we will not be introducing any exotic species </ins>into the <ins class="diffchange diffchange-inline">gut</ins>. <<ins class="diffchange diffchange-inline">br</ins>></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"> </del><<del class="diffchange diffchange-inline">li</del>><del class="diffchange diffchange-inline">Peptide displaying:</del></<del class="diffchange diffchange-inline">li</del>></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><<ins class="diffchange diffchange-inline">br</ins>><ins class="diffchange diffchange-inline"><b>References.</ins></<ins class="diffchange diffchange-inline">strong</ins>><ins class="diffchange diffchange-inline"><br></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><p>Collins M. D. and Gibson G. R.. 1999. Probiotics, prebiotics and synbiotics: Approaches for modulating the microbial ecology of the gut. Am. J. Clin. Nutr. 69:1052S–1057S </ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p><del class="diffchange diffchange-inline">Without the outer membrane</del>, <del class="diffchange diffchange-inline"><em>Bacillus subtilis</em> is ideal for peptide displaying</del>. <del class="diffchange diffchange-inline">The cell wall of <em>B</del>. <del class="diffchange diffchange-inline">subtilis</em> is the surface of the bacterium and contains approximately 9% of the total protein</del>. <del class="diffchange diffchange-inline">The discovery and study in cell wall binding modules from cell wall bound proteins provides an attractive tool for surface display of peptide</del>. <del class="diffchange diffchange-inline">(Pooley et al</del>. <del class="diffchange diffchange-inline">1996) Aiming to display the tumor-binding peptide, RPMrel, on bacteria surface, we decided to take advantage of the cell wall displaying system, LytC, in <em></del>Bacillus subtilis<del class="diffchange diffchange-inline"></em></del>. <del class="diffchange diffchange-inline">Thus, the ease of peptide displaying serves as one of our reasons to choose <em>B</del>. <del class="diffchange diffchange-inline">subtilis</em> as our chassis</del>. <del class="diffchange diffchange-inline"></p></del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p><ins class="diffchange diffchange-inline">Corazza G.R.</ins>, <ins class="diffchange diffchange-inline">Benati G</ins>.<ins class="diffchange diffchange-inline">, Strocchi A</ins>.<ins class="diffchange diffchange-inline">, Sorge M</ins>. <ins class="diffchange diffchange-inline">& Gasbarrini G</ins>.. <ins class="diffchange diffchange-inline">1992. Treatment with </ins>Bacillus subtilis <ins class="diffchange diffchange-inline">reduces intestinal hydrogen production in patients with gaseous symptoms</ins>. <ins class="diffchange diffchange-inline">Current therapeutic research</ins>. <ins class="diffchange diffchange-inline">52</ins>.<ins class="diffchange diffchange-inline">1: 144-151</ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"> </del><<del class="diffchange diffchange-inline">li</del>><del class="diffchange diffchange-inline">Normal flora in gut</del>:<del class="diffchange diffchange-inline"></li></del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><<ins class="diffchange diffchange-inline">p</ins>><ins class="diffchange diffchange-inline">Edberg, S.C.. 1991. US EPA human health assessment</ins>: <ins class="diffchange diffchange-inline">Bacillus subtilis. Unpublished, U.S. Environmental Protection Agency, Washington, D.C. </ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p><del class="diffchange diffchange-inline"><em>B. subtilis</em> not only exists widely in the nature, but also contributes to part of the normal flora in gut. (Collin and Gibson 1999) Regarded as a probiotic, it has been proved to have positive effects on patients suffering from gaseous symptom (Corazza et al. 1992). Using <em>B. subtilis</em> as our chassis, in theory, will not introduce any exotic species into gut either. </p></ol></del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>Pooley H. M., Merchante R. and Karamata D.. 1996. Overall protein content and induced enzyme components of the periplasm of Bacillus subtilis. Microb. Drug Resist.2:9–15.<ins class="diffchange diffchange-inline"> </ins></<ins class="diffchange diffchange-inline">p</ins>></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"><p><strong>References: </strong><br /></del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"> Edberg,S.C.. 1991. US EPA human health assessment: <i>Bacillus subtilis</i>. Unpublished, U.S. Environmental Protection Agency, Washington, D.C.<br /></del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"> </del>Pooley H. M., Merchante R. and Karamata D.. 1996. Overall <del class="diffchange diffchange-inline"> </del>protein content and induced enzyme components of the periplasm of <del class="diffchange diffchange-inline"><em></del>Bacillus subtilis<del class="diffchange diffchange-inline"></em></del>. <del class="diffchange diffchange-inline"><em></del>Microb. Drug Resist<del class="diffchange diffchange-inline"></em></del>.2:9–15.<<del class="diffchange diffchange-inline">br </del>/></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"> Collins M. D. and Gibson G. R.. 1999. Probiotics, prebioticsand synbiotics: Approaches for modulating the microbial ecology of the gut. <i>Am. J. Clin. Nutr.</i> 69:1052S–1057S<br /></del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"> Corazza G.R., Benati G., Strocchi A., Sorge M. &amp; Gasbarrini G.. 1992. Treatment with <i>Bacillus subtilis</i> reduces intestinal hydrogen production in patients with gaseous symptoms. <em>Current therapeutic research. </em>52.1: 144-151</p></del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"> </del></div></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
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</table>Scarimhttp://2012.igem.org/wiki/index.php?title=Team:HKUST-Hong_Kong/Design_Chassis&diff=207809&oldid=prevScarim at 17:58, 26 September 20122012-09-26T17:58:46Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>We chose <i>Bacillus subtilis</i> - a model Gram-positive bacterial species often used in laboratory study - as our chassis. It is ideal as a base for designing mechanisms to fulfill our project aims for reasons we mention below.<br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>We chose <i>Bacillus subtilis</i> - a model Gram-positive bacterial species often used in laboratory study - as our chassis. It is ideal as a base for designing mechanisms to fulfill our project aims for reasons we mention below.<br></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><<del class="diffchange diffchange-inline">p</del>><b>Safety.</b></p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><<ins class="diffchange diffchange-inline">br</ins>><b>Safety.</b></p></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>The safety issues concerning synthetic biology are numerous, particularly when it comes to the area of medical treatment. To reduce the chance of recombinant bacteria causing harm, synthetic biologists try to develop and utilize solutions that minimize the potential hazards of their products. Regarding safety issues, we the HKUST iGEM 2012 team chose to use <i>B. subtilis</i>, a non-pathogenic chassis.<del class="diffchange diffchange-inline"></p></del><br></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>The safety issues concerning synthetic biology are numerous, particularly when it comes to the area of medical treatment. To reduce the chance of recombinant bacteria causing harm, synthetic biologists try to develop and utilize solutions that minimize the potential hazards of their products. Regarding safety issues, we the HKUST iGEM 2012 team chose to use <i>B. subtilis</i>, a non-pathogenic chassis.<br></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><br<del class="diffchange diffchange-inline">><p</del>>Low degree of virulence.</p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><br>Low degree of virulence.</p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>As a natural member of the human gut microbiome, <i>B. subtilis</i> is considered to be non-pathogenic to humans. There are very few cases of humans being infected by <i>B. subtilis</i>, and of those who have been infected the vast majority had severe immune deficiency. According to Edberg (1991), <i>B. subtilis</i> does not produce significant quantities of extracellular enzymes or possess other virulence factors that would predispose it to cause infection. In other word, <i>B. subtilis</i> possesses low virulence to humans and has a low risk of adverse effects to human health. </p><br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>As a natural member of the human gut microbiome, <i>B. subtilis</i> is considered to be non-pathogenic to humans. There are very few cases of humans being infected by <i>B. subtilis</i>, and of those who have been infected the vast majority had severe immune deficiency. According to Edberg (1991), <i>B. subtilis</i> does not produce significant quantities of extracellular enzymes or possess other virulence factors that would predispose it to cause infection. In other word, <i>B. subtilis</i> possesses low virulence to humans and has a low risk of adverse effects to human health. </p><br></div></td></tr>
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</table>Scarimhttp://2012.igem.org/wiki/index.php?title=Team:HKUST-Hong_Kong/Design_Chassis&diff=207777&oldid=prevScarim at 17:58, 26 September 20122012-09-26T17:58:00Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <h1></h1></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <h1></h1></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </div></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </div></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>We chose <i>Bacillus subtilis</i> - a model Gram-positive bacterial species often used in laboratory study - as our chassis. It is ideal as a base for designing mechanisms to fulfill our project aims for reasons we mention below.<del class="diffchange diffchange-inline"></p></del><br></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>We chose <i>Bacillus subtilis</i> - a model Gram-positive bacterial species often used in laboratory study - as our chassis. It is ideal as a base for designing mechanisms to fulfill our project aims for reasons we mention below.<br></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p><b>Safety</b></p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p><b>Safety<ins class="diffchange diffchange-inline">.</ins></b></p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>The safety issues concerning synthetic biology are numerous, particularly when it comes to the area of medical treatment. To reduce the chance of recombinant bacteria causing harm, synthetic biologists try to develop and utilize solutions that minimize the potential hazards of their products. Regarding safety issues, we the HKUST iGEM 2012 team chose to use <i>B. subtilis</i>, a non-pathogenic chassis.</p><br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>The safety issues concerning synthetic biology are numerous, particularly when it comes to the area of medical treatment. To reduce the chance of recombinant bacteria causing harm, synthetic biologists try to develop and utilize solutions that minimize the potential hazards of their products. Regarding safety issues, we the HKUST iGEM 2012 team chose to use <i>B. subtilis</i>, a non-pathogenic chassis.</p><br></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;"><br></del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;"> <li>Low degree of virulence:</li></del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p><del class="diffchange diffchange-inline">Inhabiting gut as </del>a <del class="diffchange diffchange-inline">part </del>of <del class="diffchange diffchange-inline">normal flora</del>, <<del class="diffchange diffchange-inline">em</del>><del class="diffchange diffchange-inline">Bacillus </del>subtilis</<del class="diffchange diffchange-inline">em</del>> is considered to be <del class="diffchange diffchange-inline">a </del>non-pathogenic <del class="diffchange diffchange-inline">organism </del>to <del class="diffchange diffchange-inline">human</del>. There <del class="diffchange diffchange-inline">is few case that human beings </del>are infected by <<del class="diffchange diffchange-inline">em</del>>B. subtilis</<del class="diffchange diffchange-inline">em</del>>. According <del class="diffchange diffchange-inline"> </del>to Edberg (<del class="diffchange diffchange-inline">Edberg </del>1991), <<del class="diffchange diffchange-inline">em</del>>B. subtilis</<del class="diffchange diffchange-inline">em</del>> does not produce significant quantities of extracellular enzymes or possess other virulence factors that would predispose it to cause infection. In other word, <<del class="diffchange diffchange-inline">em</del>>B. subtilis</<del class="diffchange diffchange-inline">em</del>> possesses low virulence to <del class="diffchange diffchange-inline">human </del>and <del class="diffchange diffchange-inline">presents </del>low risk of adverse effects to human health. <del class="diffchange diffchange-inline"> </del></p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><br></ins><p><ins class="diffchange diffchange-inline">Low degree of virulence.</p></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><p>As </ins>a <ins class="diffchange diffchange-inline">natural member </ins>of <ins class="diffchange diffchange-inline">the human gut microbiome</ins>, <<ins class="diffchange diffchange-inline">i</ins>><ins class="diffchange diffchange-inline">B. </ins>subtilis</<ins class="diffchange diffchange-inline">i</ins>> is considered to be non-pathogenic to <ins class="diffchange diffchange-inline">humans</ins>. There are <ins class="diffchange diffchange-inline">very few cases of humans being </ins>infected by <<ins class="diffchange diffchange-inline">i</ins>>B. subtilis</<ins class="diffchange diffchange-inline">i</ins>><ins class="diffchange diffchange-inline">, and of those who have been infected the vast majority had severe immune deficiency</ins>. According to Edberg (1991), <<ins class="diffchange diffchange-inline">i</ins>>B. subtilis</<ins class="diffchange diffchange-inline">i</ins>> does not produce significant quantities of extracellular enzymes or possess other virulence factors that would predispose it to cause infection. In other word, <<ins class="diffchange diffchange-inline">i</ins>>B. subtilis</<ins class="diffchange diffchange-inline">i</ins>> possesses low virulence to <ins class="diffchange diffchange-inline">humans </ins>and <ins class="diffchange diffchange-inline">has a </ins>low risk of adverse effects to human health. </p<ins class="diffchange diffchange-inline">><br</ins>></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"> </del><<del class="diffchange diffchange-inline">li</del>>Establishment of <del class="diffchange diffchange-inline"> Integration </del>vectors<del class="diffchange diffchange-inline">:</del></<del class="diffchange diffchange-inline">li</del>></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><<ins class="diffchange diffchange-inline">br><p</ins>>Establishment of <ins class="diffchange diffchange-inline">integration </ins>vectors<ins class="diffchange diffchange-inline">.</ins></<ins class="diffchange diffchange-inline">p</ins>></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p align="left">The discovery of natural integration in <em>Bacillus subtilis</em> raised great attention in the field of molecular genetics in 1978. Due to its natural integration, a series of integration vectors for <em>B. subtilis</em> were designed later for different purposes. In our project, we employed an integration vector not only for its stability in <em>B. subtilis</em>, but also its advantages in safety. The employment of integration vector to some extent minimizes the risk of antibiotic resistance spreading within the normal flora in gut. In addition, since the exposure of BMP2 to normal tissues can induce adverse effect, integrating target genes into genome can reduce the chance of spreading of <i>Bmp2</i> gene through horizontal gene transfer and avoid non-specific drug release in gut. </p></ol></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p align="left">The discovery of natural integration in <em>Bacillus subtilis</em> raised great attention in the field of molecular genetics in 1978. Due to its natural integration, a series of integration vectors for <em>B. subtilis</em> were designed later for different purposes. In our project, we employed an integration vector not only for its stability in <em>B. subtilis</em>, but also its advantages in safety. The employment of integration vector to some extent minimizes the risk of antibiotic resistance spreading within the normal flora in gut. In addition, since the exposure of BMP2 to normal tissues can induce adverse effect, integrating target genes into genome can reduce the chance of spreading of <i>Bmp2</i> gene through horizontal gene transfer and avoid non-specific drug release in gut. </p></ol></div></td></tr>
</table>Scarimhttp://2012.igem.org/wiki/index.php?title=Team:HKUST-Hong_Kong/Design_Chassis&diff=207672&oldid=prevScarim at 17:55, 26 September 20122012-09-26T17:55:20Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>We chose <i>Bacillus subtilis</i> - a model Gram-positive bacterial species often used in laboratory study - as our chassis. It is ideal as a base for designing mechanisms to fulfill our project aims for reasons we mention below.</p><br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>We chose <i>Bacillus subtilis</i> - a model Gram-positive bacterial species often used in laboratory study - as our chassis. It is ideal as a base for designing mechanisms to fulfill our project aims for reasons we mention below.</p><br></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p><b>Safety</b></p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p><b>Safety</b></p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>The safety issues concerning synthetic biology are numerous, particularly when it comes to the area of medical treatment. To reduce the chance of recombinant bacteria causing harm, synthetic biologists try to develop and utilize solutions that minimize the potential hazards of their products. Regarding safety issues, we the HKUST iGEM 2012 team chose to use <i>B. subtilis</i>, a non-pathogenic chassis.</p><br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>The safety issues concerning synthetic biology are numerous, particularly when it comes to the area of medical treatment. To reduce the chance of recombinant bacteria causing harm, synthetic biologists try to develop and utilize solutions that minimize the potential hazards of their products. Regarding safety issues, we the HKUST iGEM 2012 team chose to use <i>B. subtilis</i>, a non-pathogenic chassis.</p><br></div></td></tr>
</table>Scarimhttp://2012.igem.org/wiki/index.php?title=Team:HKUST-Hong_Kong/Design_Chassis&diff=207644&oldid=prevScarim at 17:54, 26 September 20122012-09-26T17:54:43Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <h1></h1></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <h1></h1></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </div></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </div></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>We chose Bacillus subtilis - a model Gram-positive bacterial species often used in laboratory study - as our chassis. It is ideal as a base for designing mechanisms to fulfill our project aims for reasons we mention below.</p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>We chose <ins class="diffchange diffchange-inline"><i></ins>Bacillus subtilis<ins class="diffchange diffchange-inline"></i> </ins>- a model Gram-positive bacterial species often used in laboratory study - as our chassis. It is ideal as a base for designing mechanisms to fulfill our project aims for reasons we mention below.</p><<ins class="diffchange diffchange-inline">br</ins>></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><<del class="diffchange diffchange-inline">ol></del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"> <li>Safety</li</del>></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>Safety issues concerning synthetic biology are <del class="diffchange diffchange-inline">always challenged</del>, <del class="diffchange diffchange-inline">especially </del>when it comes to the area of medical treatment<del class="diffchange diffchange-inline">. From bacteria hosts to biowastes, the public has never ceased to raise worries</del>. To <del class="diffchange diffchange-inline">ease concerns from </del>the <del class="diffchange diffchange-inline">public</del>, <del class="diffchange diffchange-inline">researchers </del>try <del class="diffchange diffchange-inline">their best </del>to minimize the <del class="diffchange diffchange-inline">biohazardous effect involved in every step </del>of their <del class="diffchange diffchange-inline">experiments</del>. Regarding safety issues, we<del class="diffchange diffchange-inline">, 2012 </del>HKUST iGEM team<del class="diffchange diffchange-inline">, therefore </del>chose to use <<del class="diffchange diffchange-inline">em</del>>B. subtilis</<del class="diffchange diffchange-inline">em</del>>, a non-pathogenic chassis <del class="diffchange diffchange-inline">in our project</del>.</p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><br></ins></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><<del class="diffchange diffchange-inline">ol</del>></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p<ins class="diffchange diffchange-inline">><b</ins>>Safety<ins class="diffchange diffchange-inline"></b></p></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><p>The safety </ins>issues concerning synthetic biology are <ins class="diffchange diffchange-inline">numerous</ins>, <ins class="diffchange diffchange-inline">particularly </ins>when it comes to the area of medical treatment. To <ins class="diffchange diffchange-inline">reduce </ins>the <ins class="diffchange diffchange-inline">chance of recombinant bacteria causing harm</ins>, <ins class="diffchange diffchange-inline">synthetic biologists </ins>try to <ins class="diffchange diffchange-inline">develop and utilize solutions that </ins>minimize the <ins class="diffchange diffchange-inline">potential hazards </ins>of their <ins class="diffchange diffchange-inline">products</ins>. Regarding safety issues, we <ins class="diffchange diffchange-inline">the </ins>HKUST iGEM <ins class="diffchange diffchange-inline">2012 </ins>team chose to use <<ins class="diffchange diffchange-inline">i</ins>>B. subtilis</<ins class="diffchange diffchange-inline">i</ins>>, a non-pathogenic chassis.</p<ins class="diffchange diffchange-inline">><br</ins>></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><<ins class="diffchange diffchange-inline">br</ins>></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <li>Low degree of virulence:</li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <li>Low degree of virulence:</li></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
</table>Scarimhttp://2012.igem.org/wiki/index.php?title=Team:HKUST-Hong_Kong/Design_Chassis&diff=207036&oldid=prevScarim at 17:39, 26 September 20122012-09-26T17:39:25Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <h1></h1></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <h1></h1></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </div></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </div></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p><del class="diffchange diffchange-inline">Considering the requirements of our project, <em></del>Bacillus <del class="diffchange diffchange-inline"> </del>subtilis<del class="diffchange diffchange-inline"></em>, </del>a Gram-positive <del class="diffchange diffchange-inline">model organism </del>in <del class="diffchange diffchange-inline">lab </del>study<del class="diffchange diffchange-inline">, </del>is <del class="diffchange diffchange-inline">chosen </del>to <del class="diffchange diffchange-inline">be the chassis in </del>our project. <del class="diffchange diffchange-inline">Its ideality can be illustrated in the following aspects: </del></p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p><ins class="diffchange diffchange-inline">We chose </ins>Bacillus subtilis <ins class="diffchange diffchange-inline">- </ins>a <ins class="diffchange diffchange-inline">model </ins>Gram-positive <ins class="diffchange diffchange-inline">bacterial species often used </ins>in <ins class="diffchange diffchange-inline">laboratory </ins>study <ins class="diffchange diffchange-inline">- as our chassis. It </ins>is <ins class="diffchange diffchange-inline">ideal as a base for designing mechanisms </ins>to <ins class="diffchange diffchange-inline">fulfill </ins>our project <ins class="diffchange diffchange-inline">aims for reasons we mention below</ins>.</p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><ol></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><ol></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <li>Safety</li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <li>Safety</li></div></td></tr>
</table>Scarim