Team:Hong Kong HKUST/riboregulator/future work

From 2014.igem.org

(Difference between revisions)
 
(15 intermediate revisions not shown)
Line 1: Line 1:
-
<html>
+
{{Team:Hong_Kong_HKUST/shell|
-
<head>
+
<html><head>
-
+
-
<link rel="stylesheet" href="https://2014.igem.org/wiki/index.php?title=Template:Team:Hong_Kong_HKUST/anti-main.css&action=raw&ctype=text/css" type="text/css" >
+
-
<link rel="stylesheet" href="https://2014.igem.org/wiki/index.php?title=Template:Team:Hong_Kong_HKUST/indexpage.css&action=raw&ctype=text/css" type="text/css" >
+
-
<script src="http://ajax.aspnetcdn.com/ajax/jQuery/jquery-1.11.1.min.js"></script>
+
-
<script src="https://2014.igem.org/wiki/index.php?title=Template:Team:Hong_Kong_HKUST/access-menu.js&action=raw&ctype=text/css"></script>
+
-
+
-
</head>
+
-
+
-
<body>
+
-
<nav role="navigation">
+
-
<ul class="access-menu">
+
-
<li><a href="#">Home</a></li>
+
-
<li>
+
-
<a href="https://2014.igem.org/Team:Hong_Kong_HKUST/pneumosensor">Pneumosensor</a>
+
-
<ul class="access-submenu">
+
-
<li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/pneumosensor/module_one">Sensing</a></li>
+
-
<li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/pneumosensor/module_two">Expressing</a></li>
+
-
<li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/pneumosensor/parts">Parts</a></li>
+
-
<li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/pneumosensor/data">Data</a></li>
+
-
<li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/pneumosensor/characterization">Characterization</a></li>
+
-
<li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/pneumosensor/results">Results</a></li>
+
-
<li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/pneumosensor/future_work">Future Work</a></li>
+
-
</ul>
+
-
</li>
+
-
<li>
+
-
<a href="https://2014.igem.org/Team:Hong_Kong_HKUST/riboregulator">Riboregulator</a>
+
-
<ul class="access-submenu">
+
-
<li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/riboregulator/CR_TA_Feature_Page">Feature Page</a></li>
+
-
<li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/riboregulator/RNA_devices_catalog">Catalog Page</a></li>
+
-
<li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/riboregulator/parts">Parts</a></li>
+
-
<li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/riboregulator/data">Data</a></li>
+
-
<li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/riboregulator/characterization">Characterization</a></li>
+
-
<li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/riboregulator/results">Results</a></li>
+
-
<li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/riboregulator/future_work">Future Work</a></li>
+
-
</ul>
+
-
</li>
+
-
<li>
+
-
<a href="https://2014.igem.org/Team:Hong_Kong_HKUST/human_practice">Human Practice</a>
+
-
<ul class="access-submenu">
+
-
<li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/human_practice/start-up_kit">Start-up Kit</a></li>
+
-
<li class= "indent_list"><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/human_practice/start-up_kit/handbook" >--Handbook</a></li>
+
-
<li class= "indent_list"><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/human_practice/start-up_kit/report" >--Report</a></li>
+
-
<li class= "indent_list"><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/human_practice/start-up_kit/database" >--Database</a></li>
+
-
<li class= "indent_list"><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/human_practice/start-up_kit/interview" >--Interview</a></li>
+
-
<li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/human_practice/outreach">Outreach</a></li>
+
-
<li class= "indent_list"><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/human_practice/outreach/Workshop" >--Workshop</a></li>
+
-
<li class= "indent_list"><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/human_practice/outreach/talks" >--Talk</a></li>
+
-
<li class= "indent_list"><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/human_practice/outreach/isf_academy" >--ISF Academy</a></li>
+
-
<li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/human_practice/safety_and_ethics">Safety and Ethics</a></li>
+
-
</ul>
+
-
</li>
+
-
<li>
+
-
<a href="https://2014.igem.org/Team:Hong_Kong_HKUST/team">Team</a>
+
-
<ul class="access-submenu">
+
-
<li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/team/members">Members</a></li>
+
-
<li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/team/advisers">Advisers</a></li>
+
-
<li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/team/instructors">Instructors</a></li>
+
-
<li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/team/attribution">Attributions</a></li>
+
-
<li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/team/acknowledgement">Acknowledgement</a></li>
+
-
<li><a href="https://igem.org/Team.cgi?year=2014">Official Team Profile</a></li>
+
-
</ul>
+
-
</li>
+
-
<li>
+
-
<a href="https://2014.igem.org/Team:Hong_Kong_HKUST/wetlab">Wetlab</a>
+
-
<ul class="access-submenu">
+
-
<li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/wetlab/notebook">Notebook</a></li>
+
-
<li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/wetlab/protocols">Protocols</a></li>
+
-
<li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/wetlab/safety">Safety</a></li>
+
-
</ul>
+
-
</li>
+
-
<li>
+
-
<a href="https://2014.igem.org/Team:Hong_Kong_HKUST/Achievements">Achievements</a>
+
-
<ul class="access-submenu">
+
-
<li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/acheivement/medal_requirement">Medal Requirements</a></li>
+
-
<li><a href="https://2014.igem.org/Team:Hong_Kong_HKUST/acheivement/deliverable">Deliverable</a></li>
+
-
</ul>
+
-
</li>
+
-
<li class="access_logo">
+
-
<a href="https://2014.igem.org/Main_Page"><img src= "https://static.igem.org/mediawiki/2014/5/55/Hkust_logo.gif"></a>
+
-
</li>
+
-
<li class="access_logo">
+
-
<a href="http://www.ust.hk/"><img src= "https://static.igem.org/mediawiki/igem.org/6/60/Igemlogo_300px.png"></a>
+
-
</li>
+
-
+
-
</ul>
+
</head></html>
-
</nav>
+
|
-
<!-- ================ do not touch any thing above this, dont even think about it =========================-->
+
<html><body>
-
+
<div id="content_container">
-
+
-
+
-
+
-
<div id="content_container">
+
-
<div class= "banner_area">
+
-
<img src= 'https://static.igem.org/mediawiki/2014/archive/3/3c/20140930022303!HKUST_2014_pneumosensor_banner.jpg' />
+
-
<!--banner should be larger and height should be more than 400px-->
+
-
</div>
+
-
+
<div id="description_area">
<div id="description_area">
<h2>Riboregulator Future Work</h2>
<h2>Riboregulator Future Work</h2>
Line 104: Line 11:
<!-- one row of content , two column one picture right-->
<!-- one row of content , two column one picture right-->
-
<div class='content_1'><h3> Lysis Module </h3>
+
<div class='content_1'><h3>Catalog Page</h3>
<table class="content_table" align= "center" >
<table class="content_table" align= "center" >
-
<tr class= "content_row">
+
<tr class= "content_row" >
-
<td class= "content_cell">
+
<td class= "content_cell" colspan= "2">
-
<div class= "content_area_one_row">
+
<div class= "content_area_two_row">
-
<p>
+
<p>
-
Our team designed but was unable to test this module due to the limitation of not being able to work directly with <i>Streptococcus
+
                                                Since the number of regulatory RNAs in part registry have been increasing over the years and they have not been well curated, our team wishes to solve this problem by creating a list of category tags and guidelines so that one can search these parts efficiently. Therefore, we will be continuing our work in testing and characterizing the regulatory RNAs for the ease of the synthetic biology community's use in the future.
-
Pneumoniae</i> (Biosafety level 2) in our lab. This module proposes to kill <i>Streptococcus Pneumoniae</i> upon detection when coupled
+
        </p><br><br>
-
with the detection and regulation modules by releasing specific bacteriophage lytic enzymes, Cpl-1 and Pal.
+
<p> Please visit our <a href= "https://2014.igem.org/Team:Hong_Kong_HKUST/riboregulator/RNA_devices_catalog">prototype of the catalog page</a> and stay tuned with our updates.</p>
-
</p>
+
-
<p>
+
-
<br>
+
-
<br>
+
-
Amidase (PAL)- cleaves the peptidoglycan between N-acetylmuramic acid and L-alanine
+
-
<br>
+
-
<br>
+
-
Lysozyme (CPl-1)- cleaves the glycosidic bond between N-acetylmuramic acid and N-acetylglucosamine
+
-
<br>
+
-
<br>
+
-
</p>
+
</div>
</div>
</td>
</td>
-
+
</tr>
-
<td class= "content_cell">
+
</table>
-
<div class= "content_area_one_row">
+
</div>
-
<div class="content_image">
+
<div class='content_1'><h3>Re-characterization of Riboregulator under different genetic context</h3>
-
<img src= "https://static.igem.org/mediawiki/2014/thumb/d/d6/HKUST_iGEM_2014_FutureWork1.jpg/800px-HKUST_iGEM_2014_FutureWork1.jpg"/>
+
<table class="content_table" align= "center" >
-
<h5>Fig ? . The lysis Module.</h5>
+
-
</div>
+
-
<p> The enzymes are tagged with osmY (Washington 2012) via a linker to be exported out of <i>Escherichia coli</i>. Both enzymes have
+
-
very different N-terminal catalytic sites and share a similar C-terminal cell wall attachment site, which binds to choline in both cases.
+
-
Cleavage with either of these enzymes results in a weakening in the cell wall, which leads to the externalization of the cytoplasmic
+
-
membrane and ultimate lysis of <i>S. pneumoniae</i>.
+
-
</p>
+
-
</div>
+
-
</td>
+
-
+
<tr class= "content_row" >
<tr class= "content_row" >
<td class= "content_cell" colspan= "2">
<td class= "content_cell" colspan= "2">
<div class= "content_area_two_row">
<div class= "content_area_two_row">
<p>
<p>
-
<u>References</u>
+
                                                The characterization result of riboregulators were out of our expectation - the taRNAs were unable to efficiently unlock their cognate crRNAs. We now realized that our genetic context used was different from how riboregulators were characterized initially. Expression of TAs were supposed to be driven from high copy plasmids while CRs were supposed to be driven from low copy vectors. Thus, we will progressively change the conditions used to characterize the constructs into a new genetic context. We hope that we can replicate others' results soon.</p><br><br>
-
<br>
+
-
<br>
+
-
J.M. Loeffler et al &quot;Rapid Killing of <i>Streptococcus pneumoniae</i> with a Bacteriophage Cell Wall Hydrolase&quot; 2001
+
-
<br>
+
-
<br>
+
-
J.M. Loeffler et al &quot;Phage Lytic Enzyme Cpl-1 as a Novel Antimicrobial for Pneumococcal Bacteremia&quot; 2003
+
-
<br>
+
-
<br>
+
-
J.M. Loeffler et al &quot;Synergistic Lethal Effect of a Combination of Phage Lytic Enzymes with Different Activities on Penicillin-Sensitive
+
-
and -Resistant <i>Streptococcus pneumoniae</i> Strains&quot; 2003
+
-
</p>
+
</div>
</div>
</td>
</td>
Line 165: Line 40:
</table>
</table>
</div>
</div>
-
<!-- one row of content , two column one picture right-->
+
<div class='content_1'><h3>Characterization of Arabinose inducible pBad/araC promoter (<a href="http://parts.igem.org/Part:BBa_I0500">BBa_I0500</a>) in high copy plasmid</h3>
-
+
<table class="content_table" align= "center" >
-
<hr>
+
<tr class= "content_row" >
-
+
<td class= "content_cell" colspan= "2">
-
<table class= "site_map_table" align= "center">
+
<div class= "content_area_two_row">
-
<tr>
+
<p>We observed a transfer function with an all-or-none response in the pBad/araC promoter on a low copy plasmid during measurement, which matched the experimental result from that of iGEM 2011 Cambridge team, who also used the same genetic context. We are now speculating that the discrepancy between results from iGEM 2011 Cambridge and Groningen was a result due to different copy number plasmids used in carrying the promoter. However, due to lack of sufficient time and manpower, we were unable to test that out in time. Our future work will involve resumption of this characterization, and hopefully, we can explain the discrepancy observed through a mathematical model.</p><br><br>
-
<td class= 'site_map_column'><h4><a href="https://2014.igem.org/Team:Hong_Kong_HKUST">Home</a></h4>
+
</div>
-
<ul class= 'site_list'>
+
</td>
-
<li class='site_link'><a href="">Pneumosensor</a></li>
+
</tr>
-
<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>
</tr>
-
+
</div>
</table>
</table>
</div>
</div>
-
+
</div>
-
+
</body></html>
-
</body>
+
}}
-
</html>
+

Latest revision as of 23:49, 17 October 2014



Riboregulator Future Work

Catalog Page

Since the number of regulatory RNAs in part registry have been increasing over the years and they have not been well curated, our team wishes to solve this problem by creating a list of category tags and guidelines so that one can search these parts efficiently. Therefore, we will be continuing our work in testing and characterizing the regulatory RNAs for the ease of the synthetic biology community's use in the future.



Please visit our prototype of the catalog page and stay tuned with our updates.

Re-characterization of Riboregulator under different genetic context

The characterization result of riboregulators were out of our expectation - the taRNAs were unable to efficiently unlock their cognate crRNAs. We now realized that our genetic context used was different from how riboregulators were characterized initially. Expression of TAs were supposed to be driven from high copy plasmids while CRs were supposed to be driven from low copy vectors. Thus, we will progressively change the conditions used to characterize the constructs into a new genetic context. We hope that we can replicate others' results soon.



Characterization of Arabinose inducible pBad/araC promoter (BBa_I0500) in high copy plasmid

We observed a transfer function with an all-or-none response in the pBad/araC promoter on a low copy plasmid during measurement, which matched the experimental result from that of iGEM 2011 Cambridge team, who also used the same genetic context. We are now speculating that the discrepancy between results from iGEM 2011 Cambridge and Groningen was a result due to different copy number plasmids used in carrying the promoter. However, due to lack of sufficient time and manpower, we were unable to test that out in time. Our future work will involve resumption of this characterization, and hopefully, we can explain the discrepancy observed through a mathematical model.



Home

Pneumosensor

Riboregulator

Human Practice

Team

WetLab

Achievement