|
|
| (5 intermediate revisions not shown) |
| Line 1: |
Line 1: |
| - | {{Team:Hong_Kong_HKUST/shell|
| |
| - | <html><head>
| |
| | | | |
| - | </head></html>
| |
| - | |
| |
| - | <html><body>
| |
| - | <div id="content_container">
| |
| - | <hr>
| |
| - |
| |
| - |
| |
| - | <!-- one row of content , two column-->
| |
| - | <div class='content_1'><h3>Overview</h3>
| |
| - | <table class="content_table" align= "center" valign= "top">
| |
| - | <tr class= "content_row">
| |
| - | <td class= "content_cell">
| |
| - | <div class= "content_area_one_row">
| |
| - | <p>Although there is a significant number of regulatory RNAs available in the registry, a comprehensive characterization information that the iGEM community can
| |
| - | use to compare and contrast different regulatory RNAs (especially CR-TA riboregulators) is missing.Therefore we wanted to provide characterization information of
| |
| - | regulatory RNAs so teams and labs will be confident in using these devices. </p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | </table>
| |
| - | </div>
| |
| - | <!-- end of one row of content , two column-->
| |
| - |
| |
| - | <!-- one row of content , two column one picture right-->
| |
| - | <div class='content_1'><h3>Construct</h3>
| |
| - | <table class="content_table" align= "center" >
| |
| - | <tr class= "content_row">
| |
| - | <td class= "content_cell">
| |
| - | <div class= "content_area_one_row">
| |
| - | <p>In order to provide reliable characterisation data which will help iGEM teams to identify which CR-TA riboregulator works best for their projects. We have built
| |
| - | constructs like the diagram provided below:
| |
| - | (provide diagram)
| |
| - | <p>Cis-repressing part which was generated by using the Lock and Key algorithm generator from TU Delft 2009 iGEM team and followed by de novo synthesis from oligo
| |
| - | annealing. After annealing, these dsDNA was directly inserted into the vector containing <a href="http://parts.igem.org/Part:BBa_I13401">BBa_I13401</a> ( GFP reporter for RHS of library test constructs). A promoter,
| |
| - | <a href="http://parts.igem.org/Part:BBa_J23102">BBa_J23102</a> was placed upstream by the scar formation of cis-repressing part therefore cis-repressing part can be
| |
| - | constitutively transcribed.Trans-activating part was produced by the same method as cis-repressing part but an inducible promoter, P<sub>bad</sub>, <a href="http://parts.igem.org/Part:BBa_I13401">BBa_I0500 </a>and double terminator,
| |
| - | <a href="http://parts.igem.org/Part:BBa_BBa_B0015">BBa_B0015 </a>were used to replace <a href="http://parts.igem.org/Part:BBa_I13401">BBa_J23102 </a>and
| |
| - | <a href="http://parts.igem.org/Part:BBa_I13401">BBa_I13401 </a>respectively. A low copy plasmid, pSB3K3 was used for GFP expression measurements. For the control of our characterisation, construct containing only RBS,
| |
| - | instead of CR and another construct missing TA were built. To obtain the fluorescence level, autofluorescence had to be subtracted so bacteria containing pSB3K3-E0240
| |
| - | plasmid were used. We have submitted Cis-repressing and Trans-activating parts as well as some of intermediary parts. </p>
| |
| - | </div>
| |
| - | </td>
| |
| - |
| |
| - | </tr>
| |
| - | </table>
| |
| - | </div>
| |
| - | <!-- end of one row of content , two column-->
| |
| - | <br><br>
| |
| - |
| |
| - | <!-- one row of content , two column one picture left-->
| |
| - | <div class='content_1'><h3> Results</h3>
| |
| - | <table class="content_table" align= "center" >
| |
| - |
| |
| - | <tr class= "content_row">
| |
| - | <td class= "content_cell">
| |
| - | <div class= "content_area_one_row">
| |
| - | <div class="content_image">
| |
| - |
| |
| - | </div>
| |
| - | <p>
| |
| - | <img style= "width:50%" src= "#"/>
| |
| - | <br>
| |
| - | The above bar graphs represents the level of fluorescence measured by ? and it was plotted as fluorescence/OD600 on y-axis. Culture containing bacteria was
| |
| - | incubated for X hours. (From left to right) For controls of each set of an experiment, (1) a construct without trans-activating part (2) a construct without
| |
| - | cis-repressing part but with a ribosomal binding site (3) a construct without both cis-repressing and trans-activating parts. For our experiment, a construct
| |
| - | containing both cis-repressing and trans-activating parts was used. arabinose concentration of 0%, 1% and 2.5% were used to induce the inducible promoter, P<sub>bad</sub>
| |
| - | (<a href="https://2014.igem.org/Team:Hong_Kong_HKUST/riboregulator/characterization">P<sub>bad</sub> characterisation</a>). </p>
| |
| - |
| |
| - | <br><br>
| |
| - | <img style= "width:50%" src= "#"/>
| |
| - | <br>
| |
| - | <p>Furthermore the orthogonality of five different sets of riboregulators were tested in parallel to the repression and activation strength of riboregulators.
| |
| - | The increase in fluorescence level was calculated using the fluorescence level obtained by plate reader at 0% and 2.5% of arabinose concentrations.</p>
| |
| - | </div>
| |
| - | </td>
| |
| - |
| |
| - |
| |
| - | </tr>
| |
| - | </div>
| |
| - | </table>
| |
| - | </div>
| |
| - |
| |
| - | <div class='content_1'><h3> Discussion</h3>
| |
| - | <table class="content_table" align= "center" >
| |
| - |
| |
| - | <tr class= "content_row">
| |
| - | <td class= "content_cell">
| |
| - |
| |
| - | <div class= "content_area_one_row">
| |
| - |
| |
| - | <p>
| |
| - | </p>
| |
| - | </div>
| |
| - | </td>
| |
| - |
| |
| - |
| |
| - | </tr>
| |
| - | </table>
| |
| - | </div>
| |
| - | </div>
| |
| - |
| |
| - | <footer>
| |
| - | <script>
| |
| - | $(document).ready(function(){
| |
| - |
| |
| - | //Check to see if the window is top if not then display button
| |
| - | $(window).scroll(function(){
| |
| - | if ($(this).scrollTop() > 100) {
| |
| - | $('.scrollToTop').fadeIn();
| |
| - | } else {
| |
| - | $('.scrollToTop').fadeOut();
| |
| - | }
| |
| - | });
| |
| - |
| |
| - | //Click event to scroll to top
| |
| - | $('.scrollToTop').click(function(){
| |
| - | $('html, body').animate({scrollTop : 0},800);
| |
| - | return false;
| |
| - | });
| |
| - |
| |
| - | });
| |
| - | </script>
| |
| - | <a href="#" class="scrollToTop">
| |
| - | <div style= "text-align:center" class="scrollToTop">
| |
| - | <br>
| |
| - | Back to top
| |
| - | </div>
| |
| - | </a>
| |
| - | </footer>
| |
| - | </body></html>
| |
| - | }}
| |
"
