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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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- | <li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/pneumosensor/module_two">Expressing</a></li>
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- | <li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/riboregulator/parts">Parts</a></li>
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- | <li><a href="https://igem.org/Team.cgi?year=2014">Official Team Profile</a></li>
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| + | <div class="content_1"> |
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- |
| + | <h3> Medal Requirements</h3> |
- |
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- | <div id="content_container"> | + | |
- | <div class= "banner_area">
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- | <img src= 'https://static.igem.org/mediawiki/2014/archive/3/3c/20140930022303!HKUST_2014_pneumosensor_banner.jpg' />
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| </div> | | </div> |
- | <hr>
| + | <div id="medal_reqire_container" > |
- |
| + | <div class= "each_medal" style="background-color:#bbab8c"> |
- |
| + | <div class= "img_container" ><img src="https://static.igem.org/mediawiki/2014/b/be/HKUST_Bronze_medal.png" ></div> |
- | <!--two row of content, image first row, text second row -->
| + | |
- | <div class='content_1'><h3> Module Two what'sthename Description </h3> | + | |
- | <table class="content_table" align= "center" >
| + | <div class= "medal_text"> |
- |
| + | <h3 class= "What_medal"> Bronze Medal</h3> |
- | <tr class= "content_row" >
| + | <ul> |
- | <td class= "content_cell" colspan= "2"> | + | <li>Team registered</li> |
- | <div class= "content_area_one_row"> | + | <li>Completed <a href="https://igem.org/2014_Judging_Form?id=1379">Judging form</a></li> |
- | <div class="content_image">
| + | <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> |
- | <img src= "https://static.igem.org/mediawiki/2014/thumb/d/d5/HKUST_iGEM_2014_M2abstract1.jpg/688px-HKUST_iGEM_2014_M2abstract1.jpg"/>
| + | </ul> |
- | </div>
| + | |
- | <p>Prof. Donald A. Morrison's research lab in University of Illinois at Chicago published several papers on the competence for genetic transformation
| + | |
- | 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, | + | |
- | processing, and integration. There is a link between this quorum-sensing system and the competence-specific genes, which is an alternative sigma | + | |
- | factor ComX. Two identical copies of gene (<i>comX1</i> and <i>comX2</i>) encode a competence-specific alternative sigma factor, σ<sup>x</sup>.
| + | |
- | Expression of ComX allows the transcription of many genes that are involved in transformation and specifically expressed during competence. These late
| + | |
- | genes share a conserved 8-bp sequence in their promoter regions, TACGAATA (combox) which is specifically induced by σ<sup>x</sup>-containing
| + | |
- | RNA polymerase.
| + | |
- | </p> | + | |
| </div> | | </div> |
- | </td>
| |
- | <td class= "content_cell" colspan= "2">
| |
- | <div class= "content_area_one_row">
| |
- | <p>The combox promoter is a promoter associated with initiating transcription of a class of genes (commonly referred to as late competence genes)
| |
- | coding for bacterial competence proteins and bacterial transformation in <i>Streptococcus Pneumoniae</i> strains. As described before, it contains
| |
- | a general consensus sequence of TACGAATA for recognition by its promoter-specific sigma factor X, which is a protein expressed from the early
| |
- | competence gene <i>comX</i>.
| |
- | </p>
| |
- | <p>In fact, the combox promoter can be found on many different regions within the genomic DNA of <i>Streptococcus
| |
- | Pneumoniae</i> strains. Researches and investigations have also reported variants of the combox promoter having different lengths and consensus
| |
- | sequences, but generally the range of variety has been kept small.
| |
- | </p>
| |
| </div> | | </div> |
- | </td>
| |
- | </tr>
| |
- | </table>
| |
| | | |
- | <table class="content_table" align= "center" >
| + | <div class= "each_medal" style="background-color:#c0c0c0"> |
- | <tr class= "content_row" >
| + | <div class= "img_container"><img src="https://static.igem.org/mediawiki/2014/f/f8/HKUST_Silver_medal.png" ></div> |
- | <td class= "content_cell">
| + | <div class= "medal_text "> |
- | <div class= "content_area_one_row"> | + | <h3 class= "What_medal"> Silver Medal</h3> |
- | <p>Though much information about the combox promoter is readily available nowadays, its full characterization including promoter activity, specificity, | + | <ul> |
- | sequence, as well as the biomolecular mechanism can be greatly enhanced with further investigations and experiments. As part of module II, we were | + | <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> |
- | interested in reproducing this gene circuit with all the associated genes and promoters to be combined into a single transcriptional unit.
| + | |
- | To the best of our knowledge, the genes and promoters of this gene circuit for bacterial transformation are found at different locations of the
| + | <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> |
- | long genomic DNA of many <i>Streptococcus Pneumoniae</i> strains. Despite the suggested susceptibility to leakage and other factors that may
| + | <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> |
- | hinder or interrupt the mechanism, researches have reported that the pathway was highly specific to certain environmental conditions and stress,
| + | </ul> |
- | 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
| + | |
- | the combox promoter, and also identifying the most probable length of the promoter. Characterization of the combox promoter was carried out
| + | |
- | together with the characterization of <i>comX</i> gene. Because combox promoter is highly specific to σ<sup>x</sup> for activation,
| + | |
- | genes downstream of the combox promoter will be translated only if σ<sup>x</sup> are present. Hence, by using fluorescence protein as a
| + | |
- | reporting mechanism, this <i>comX</i>-combox system could be further utilized as a specific reporter device that could be used by the iGEM
| + | |
- | community.
| + | |
- | </p> | + | |
| </div> | | </div> |
- | </td>
| |
- | <td class= "content_cell">
| |
- | <div class= "content_area_one_row">
| |
- | <div class="content_image">
| |
- | <img src= "https://static.igem.org/mediawiki/2014/thumb/1/1b/HKUST_iGEM_2014_M2abstract2.jpg/657px-HKUST_iGEM_2014_M2abstract2.jpg"/>
| |
- | </div>
| |
- | <br>
| |
- | <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
| |
- | 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 σ<sup>x</sup> and the expression of late genes as ComW contributes to the stabilization of the alternative sigma
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- | factor σ<sup>x</sup>against proteolysis and is required for full activity of σ<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.
| |
- | </p>
| |
- | <br>
| |
- | <p>Based on these findings, we would like to integrate this alternative sigma factor system from Gram-positive <i>Streptococcus pneumoniae</i>
| |
- | 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
| |
- | genes. Lastly, we combined these constructs with the combox promoter and GFP generator to check the functionality of the system, and to calculate the
| |
- | relative promoter unit of combox promoter.
| |
- | </p>
| |
| </div> | | </div> |
- | </td> | + | |
- | </tr> | + | <div class= "each_medal" style="background-color:#ffeb7f"> |
- | </table>
| + | <div class= "img_container"><img src="https://static.igem.org/mediawiki/2014/f/fe/HKUST_Gold_medal.png" ></div> |
- |
| + | <div class= "medal_text "> |
- | <div class='content_1'>
| + | <h3 class= "What_medal"> Gold Medal</h3> |
- | <table class="content_table" align= "center" >
| + | <ul> |
- | <tr class= "content_row" >
| + | <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> |
- | <td class= "content_cell" colspan= "2">
| + | |
- | <div class= "content_area_two_row"> | + | </ul> |
- | <p>
| + | |
- | <u>References</u>
| + | |
- | <br>
| + | |
- | <br>
| + | |
- | Andrew Piotrowski et al. (2009) "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" Journal of Bacteriology.
| + | |
- | <br>
| + | |
- | <br>
| + | |
- | Ping Luo et al. (2003) "Transient Association of an Alternative Sigma Factor, ComX, with RNA Polymerase during the Period of Competence for Genetic Transformation in <i>Streptococcus pneumonia</i>" Journal of Bacteriology.
| + | |
- | <br>
| + | |
- | <br>
| + | |
- | Sung CK et al. (2005) "Two distinct functions of ComW in stabilization and activation of the alternative sigma factor ComX in <i>Streptococcus pneumoniae</i>. " Journal of Bacteriology.
| + | |
- | <br>
| + | |
- | <br>
| + | |
- | Haiying Li et al. (2004) "Identification of ComW as a new component in regulation of genetic transformation in <i>Streptococcus Pneumoniae</i>" Molecular Biology.
| + | |
- | <br>
| + | |
- | <br>
| + | |
- | Myeong S.Lee et al. (1999) "Identification of a New Regulator in <i>Streptococcus Pneumoniae</i> Linking Quorum Sensing to Competence for Genetic Transformation"
| + | |
- | </p>
| + | |
| </div> | | </div> |
- | </td> | + | </div> |
- | </tr>
| + | |
- | </table>
| + | |
| </div> | | </div> |
| </div> | | </div> |
- | <hr>
| + | </div> |
- |
| + | |
- | <table class= "site_map_table" align= "center">
| + | |
- | <tr>
| + | |
- | <td class= 'site_map_column'><h4><a href="https://2014.igem.org/Team:Hong_Kong_HKUST">Home</a></h4>
| + | |
- | <ul class= 'site_list'>
| + | |
- | <li class='site_link'><a href="">Pneumosensor</a></li>
| + | |
- | <li class='site_link'><a href="">Riboregulator</a></li>
| + | |
- | <li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/human_practice">Human Practice</a></li>
| + | |
- | <li class='site_link'><a href="">Team</a></li>
| + | |
- | <li class='site_link'><a href="">WetLab</a></li>
| + | |
- | <li class='site_link'><a href="">Achievements</a></li>
| + | |
- | </ul>
| + | |
- |
| + | |
- | </td>
| + | |
- | <td class= 'site_map_column'><h4><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/pneumosensor">Pneumosensor</a></h4>
| + | |
- | <ul class= 'site_list'>
| + | |
- | <li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/pneumosensor/module_one">Sensing</a></li>
| + | |
- | <li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/pneumosensor/module_two">Expressing</a></li>
| + | |
- | <li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/pneumosensor/parts">Parts</a></li>
| + | |
- | <li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/pneumosensor/data">Data</a></li>
| + | |
- | <li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/pneumosensor/characterization">Characterization</a></li>
| + | |
- | <li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/pneumosensor/results">Results</a></li>
| + | |
- | <li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/pneumosensor/future_work">Future Work</a></li>
| + | |
- | </ul>
| + | |
- | </td>
| + | |
- | <td class= 'site_map_column'><h4><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/riboregulator">Riboregulator</a></h4>
| + | |
- | <ul class= 'site_list'>
| + | |
- | <li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/riboregulator/CR_TA_Feature_Page">Feature Page</a></li>
| + | |
- | <li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/riboregulator/RNA_devices_catalog">Catalog Page</a></li>
| + | |
- | <li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/riboregulator/parts">Parts</a></li>
| + | |
- | <li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/riboregulator/data">Data</a></li>
| + | |
- | <li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/riboregulator/characterization">Characterization</a></li>
| + | |
- | <li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/riboregulator/results">Results</a></li>
| + | |
- | <li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/riboregulator/future_work">Future Work</a></li>
| + | |
- | </ul>
| + | |
- | </td>
| + | |
- | <td class= 'site_map_column'><h4><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/human_practice">Human Practice</a></h4>
| + | |
- | <ul class= 'site_list'>
| + | |
- | <li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/human_practice/start-up_kit">Start-up Kit</a>
| + | |
- | <ul>
| + | |
- | <li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/human_practice/start-up_kit/handbook" >--Handbook</a></li>
| + | |
- | <li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/human_practice/start-up_kit/report" >--Report</a></li>
| + | |
- | <li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/human_practice/start-up_kit/database" >--Database</a></li>
| + | |
- | <li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/human_practice/start-up_kit/interview" >--Interview</a></li>
| + | |
- | </ul>
| + | |
- | </li>
| + | |
- | <li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/human_practice/outreach">Outreach</a>
| + | |
- | <ul>
| + | |
- | <li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/human_practice/outreach/Workshop" >--Workshop</a></li>
| + | |
- | <li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/human_practice/outreach/talks" >--Talk</a></li>
| + | |
- | <li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/human_practice/outreach/isf_academy" >--ISF Academy</a></li>
| + | |
- | </ul>
| + | |
- | </li>
| + | |
- | <li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/human_practice/safety_and_ethics">Safety and Ethics</a></li>
| + | |
- | </ul>
| + | |
- | </td>
| + | |
- | <td class= 'site_map_column'><h4><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/team">Team</a></h4>
| + | |
- | <ul class= 'site_list'>
| + | |
- | <li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/team/members">Members</a></li>
| + | |
- | <li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/team/advisers">Advisers</a></li>
| + | |
- | <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>
| + | |
- | <li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/team/acknowledgement">Acknowledgement</a></li>
| + | |
- | <li class='site_link'><a href="https://igem.org/Team.cgi?year=2014">Official Team Profile</a></li>
| + | |
- | </ul>
| + | |
- | </td>
| + | |
- | <td class= 'site_map_column'><h4><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/wetlab">WetLab</a></h4>
| + | |
- | <ul class= 'site_list'>
| + | |
- | <li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/wetlab/notebook">Notebook</a></li>
| + | |
- | <li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/wetlab/protocols">Protocols</a></li>
| + | |
- | <li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/wetlab/safety">Safety</a></li>
| + | |
- | </ul>
| + | |
- | </td>
| + | |
- | <td class= 'site_map_column'><h4><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/Achievements">Achievement</a></h4>
| + | |
- | <ul class= 'site_list'>
| + | |
- | <li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/acheivement/medal_requirement">Medal Requirements</a></li>
| + | |
- | <li class='site_link'><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/acheivement/deliverable">Deliverable</a></li>
| + | |
- | </ul>
| + | |
- | </td>
| + | |
- | | + | |
- | </tr>
| + | |
- |
| + | |
- | </table>
| + | |
- | </div>
| + | |
- |
| + | |
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| + | |
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