Team:Hong Kong HKUST/Achievements/medal requirement

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<a href="https://2014.igem.org/Team:Hong_Kong_HKUST/pneumosensor">Pneumosensor</a>
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<li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/pneumosensor/module_one">Sensing</a></li>
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<a href="https://2014.igem.org/Team:Hong_Kong_HKUST/Achievements">Achievements</a>
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<li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/acheivement/medal_requirement">Medal Requirements</a></li>
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<h3> Medal Requirements</h3>
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<div class='content_1'><h3> Module Two what'sthename Description </h3>
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<h3 class= "What_medal"> Bronze Medal</h3>
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  <li>Team registered</li>
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  <li>Completed <a href="https://igem.org/2014_Judging_Form?id=1379">Judging form</a></li>
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  <li>We have documented more than one new BioBrick Parts that we used in our project. This part is submitted to iGEM registry. Check out the <a href="https://2014.igem.org/Team:Hong_Kong_HKUST/pneumosensor/data">Data page</a> for more exciting details.</li>
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<img src= "https://static.igem.org/mediawiki/2014/thumb/d/d5/HKUST_iGEM_2014_M2abstract1.jpg/688px-HKUST_iGEM_2014_M2abstract1.jpg"/>
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<p>Prof. Donald A. Morrison&#39;s research lab in University of Illinois at Chicago published several papers on the competence for genetic transformation
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in <i>Streptococcus pneumoniae</i> which depends on a quorum-sensing system to control many competence-specific genes which play a role in DNA uptake,
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processing, and integration. There is a link between this quorum-sensing system and the competence-specific genes, which is an alternative sigma
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factor ComX. Two identical copies of gene (<i>comX1</i> and <i>comX2</i>) encode a competence-specific alternative sigma factor, &sigma;<sup>x</sup>.
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Expression of ComX allows the transcription of many genes that are involved in transformation and specifically expressed during competence. These late
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genes share a conserved 8-bp sequence in their promoter regions, TACGAATA (combox) which is specifically induced by &sigma;<sup>x</sup>-containing
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RNA polymerase.  
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<p>The combox promoter is a promoter associated with initiating transcription of a class of genes (commonly referred to as late competence genes)
 
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coding for bacterial competence proteins and bacterial transformation in <i>Streptococcus Pneumoniae</i> strains. As described before, it contains
 
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a general consensus sequence of TACGAATA for recognition by its promoter-specific sigma factor X, which is a protein expressed from the early
 
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competence gene <i>comX</i>.
 
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</p>
 
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<p>In fact, the combox promoter can be found on many different regions within the genomic DNA of <i>Streptococcus
 
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Pneumoniae</i> strains. Researches and investigations have also reported variants of the combox promoter having different lengths and consensus
 
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sequences, but generally the range of variety has been kept small.
 
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<h3 class= "What_medal"> Silver Medal</h3>
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<p>Though much information about the combox promoter is readily available nowadays, its full characterization including promoter activity, specificity,
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sequence, as well as the biomolecular mechanism can be greatly enhanced with further investigations and experiments. As part of module II, we were
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  <li>We have characterized <a href="http://parts.igem.org/Part:BBa_K1379004">BBa_K1379004</a>, <a href="http://parts.igem.org/Part:BBa_K1379000">BBa_K1379000</a> and <a href="http://parts.igem.org/Part:BBa_K1379001">BBa_K1379001</a> and they function as expected. Check out <a href="https://2014.igem.org/Team:Hong_Kong_HKUST/pneumosensor/characterization">CHARACTERIZATION</a> for more exciting details.</li>
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interested in reproducing this gene circuit with all the associated genes and promoters to be combined into a single transcriptional unit.  
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To the best of our knowledge, the genes and promoters of this gene circuit for bacterial transformation are found at different locations of the
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  <li>We also submit new parts to the iGEM part Registry. They are listed in <a href="https://2014.igem.org/Team:Hong_Kong_HKUST/pneumosensor/parts">Part page for Pneumosensor</a> as well as in <a href="https://2014.igem.org/Team:Hong_Kong_HKUST/riboregulator/parts">Part page for Riboregulator</a> </li>
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long genomic DNA of many <i>Streptococcus Pneumoniae</i> strains. Despite the suggested susceptibility to leakage and other factors that may
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  <li>We also consider the safety issues that we concern about our project. We interviewed expert, Read  <a href="https://2014.igem.org/Team:Hong_Kong_HKUST/human_practice/safety_and_ethics">the transcript of the interview</a>.</li>
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hinder or interrupt the mechanism, researches have reported that the pathway was highly specific to certain environmental conditions and stress,
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suggesting minimal or no leakage in the entire process. With part I of module to focusing on ComX, part II of module II focused mainly on isolating
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the combox promoter, and also identifying the most probable length of the promoter. Characterization of the combox promoter was carried out
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together with the characterization of <i>comX</i> gene. Because combox promoter is highly specific to &sigma;<sup>x</sup> for activation,
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genes downstream of the combox promoter will be translated only if &sigma;<sup>x</sup> are present. Hence, by using fluorescence protein as a  
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reporting mechanism, this <i>comX</i>-combox system could be further utilized as a specific reporter device that could be used by the iGEM
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community.  
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<img src= "https://static.igem.org/mediawiki/2014/thumb/1/1b/HKUST_iGEM_2014_M2abstract2.jpg/657px-HKUST_iGEM_2014_M2abstract2.jpg"/>
 
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<br>
 
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<p> Besides ComX, another positive factor involved in competence regulation was later found out to be ComW. The <i>comW</i> gene (SP0018) is regulated
 
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by the quorum-sensing system and is required for a high-level of competence. Coexpression of <i>comW</i> with <i>comX </i>restores the
 
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accumulation of &sigma;<sup>x</sup> and the expression of late genes as ComW contributes to the stabilization of the alternative sigma
 
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factor &sigma;<sup>x</sup>against proteolysis and is required for full activity of &sigma;<sup>x</sup> in directing transcription of late
 
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competence genes. ComW functions to act as a barrier which covers comX protein from being degraded by the ClpXP degradation enzyme. Hence, ComW
 
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will be degraded instead of ComX, and the production of ComX protein will be increased.
 
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</p>
 
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<br>
 
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<p>Based on these findings, we would like to integrate this alternative sigma factor system from Gram-positive <i>Streptococcus pneumoniae</i>
 
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into Gram-negative <i>Escherichia coli</i>. Firstly, we cloned out the <i>comX</i> and <i>comW</i> genes from the genomic DNA of <i>S. pneumoniae</i>
 
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NCTC 7465 strain. We then used BioBrick BBa_K880005 (consisting of constitutive promoter J23100 and strong RBS B0034) to drive the expression of those
 
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genes. Lastly, we combined these constructs with the combox promoter and GFP generator to check the functionality of the system, and to calculate the
 
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relative promoter unit of combox promoter.
 
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<h3 class= "What_medal"> Gold Medal</h3>
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  <li>We found out <a href="http://parts.igem.org/Part:BBa_I0500">BBa_I0500</a> P<sub>bad</sub> promoter has inconsistent sequence. We send another functional P<sub>bad</sub> promoter to Part registry; We also measure <a href ="https://2014.igem.org/Team:Hong_Kong_HKUST/riboregulator/characterization">the RPU of P<sub>bad</sub> across different strain of <i>E. coli</i></a></li>
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<u>References</u>
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<br>
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<br>
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Andrew Piotrowski et al. (2009) &quot;Competence for Genetic Transformation in <i>Streptococcus pneumoninae</i>: Termination of Activity of the Alternative Sigma Factor ComX Is Independent of Proteolysis of ComX and ComW&quot; Journal of Bacteriology.
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<br>
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<br>
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Ping Luo et al. (2003) &quot;Transient Association of an Alternative Sigma Factor, ComX, with RNA Polymerase during the Period of Competence for Genetic Transformation in <i>Streptococcus pneumonia</i>&quot; Journal of Bacteriology.
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<br>
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<br>
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Sung CK et al. (2005) &quot;Two distinct functions of ComW in stabilization and activation of the alternative sigma factor ComX in <i>Streptococcus pneumoniae</i>. &quot; Journal of Bacteriology.
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<br>
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<br>
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Haiying Li et al. (2004) &quot;Identification of ComW as a new component in regulation of genetic transformation in <i>Streptococcus Pneumoniae</i>&quot; Molecular Biology.
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<br>
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<br>
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Myeong S.Lee et al. (1999) &quot;Identification of a New Regulator in <i>Streptococcus Pneumoniae</i> Linking Quorum Sensing to Competence for Genetic Transformation&quot;
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<td class= 'site_map_column'><h4><a href="https://2014.igem.org/Team:Hong_Kong_HKUST">Home</a></h4>
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<ul class= 'site_list'>
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<li class='site_link'><a href="">Pneumosensor</a></li>
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<li class='site_link'><a href="">Riboregulator</a></li>
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<li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/human_practice">Human Practice</a></li>
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<li class='site_link'><a href="">Team</a></li>
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<li class='site_link'><a href="">WetLab</a></li>
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<li class='site_link'><a href="">Achievements</a></li>
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<td class= 'site_map_column'><h4><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/pneumosensor">Pneumosensor</a></h4>
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<li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/pneumosensor/module_one">Sensing</a></li>
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<li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/pneumosensor/module_two">Expressing</a></li>
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<li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/pneumosensor/parts">Parts</a></li>
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<li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/pneumosensor/data">Data</a></li>
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<li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/pneumosensor/characterization">Characterization</a></li>
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<li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/pneumosensor/results">Results</a></li>
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<li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/pneumosensor/future_work">Future Work</a></li>
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<td class= 'site_map_column'><h4><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/riboregulator">Riboregulator</a></h4>
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<li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/riboregulator/RNA_devices_catalog">Catalog Page</a></li>
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<li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/riboregulator/parts">Parts</a></li>
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<li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/riboregulator/data">Data</a></li>
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<li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/riboregulator/characterization">Characterization</a></li>
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<li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/riboregulator/results">Results</a></li>
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<li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/riboregulator/future_work">Future Work</a></li>
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</ul>
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</td>
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<td class= 'site_map_column'><h4><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/human_practice">Human Practice</a></h4>
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<ul class= 'site_list'>
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<li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/human_practice/start-up_kit">Start-up Kit</a>
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<ul>
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<li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/human_practice/start-up_kit/handbook" >--Handbook</a></li>
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<li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/human_practice/start-up_kit/report" >--Report</a></li>
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<li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/human_practice/start-up_kit/database" >--Database</a></li>
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<li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/human_practice/start-up_kit/interview" >--Interview</a></li>
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</ul>
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</li>
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<li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/human_practice/outreach">Outreach</a>
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<ul>
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<li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/human_practice/outreach/Workshop" >--Workshop</a></li>
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<li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/human_practice/outreach/talks" >--Talk</a></li>
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<li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/human_practice/outreach/isf_academy" >--ISF Academy</a></li>
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</ul>
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</li>
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<li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/human_practice/safety_and_ethics">Safety and Ethics</a></li>
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</ul>
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</td>
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<td class= 'site_map_column'><h4><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/team">Team</a></h4>
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<ul class= 'site_list'>
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<li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/team/members">Members</a></li>
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<li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/team/advisers">Advisers</a></li>
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<li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/team/instructors">Instructors</a></li>
+
-
<li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/team/attribution">Attributions</a></li>
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-
<li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/team/acknowledgement">Acknowledgement</a></li>
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<li class='site_link'><a href="https://igem.org/Team.cgi?year=2014">Official Team Profile</a></li>
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</ul>
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</td>
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-
<td class= 'site_map_column'><h4><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/wetlab">WetLab</a></h4>
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<ul class= 'site_list'>
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-
<li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/wetlab/notebook">Notebook</a></li>
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-
<li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/wetlab/protocols">Protocols</a></li>
+
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<li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/wetlab/safety">Safety</a></li>
+
-
</ul>
+
-
</td>
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<td class= 'site_map_column'><h4><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/Achievements">Achievement</a></h4>
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<ul class= 'site_list'>
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<li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/acheivement/medal_requirement">Medal Requirements</a></li>
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<li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/acheivement/deliverable">Deliverable</a></li>
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</ul>
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</td>
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Latest revision as of 23:48, 17 October 2014



Medal Requirements

Bronze Medal

  • Team registered
  • Completed Judging form
  • We have documented more than one new BioBrick Parts that we used in our project. This part is submitted to iGEM registry. Check out the Data page for more exciting details.

Silver Medal

Gold Medal

Home

Pneumosensor

Riboregulator

Human Practice

Team

WetLab

Achievement