IGEM:IMPERIAL/2008/New/Chassis 2: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
No edit summary
 
(69 intermediate revisions by 5 users not shown)
Line 1: Line 1:
{{Imperial/StartPage}}__NOTOC__
{{Imperial/StartPage2}}
 
<br><br>
=Chassis 2 - Other pros, cons and information...=
&nbsp; <font size=6px color=#E5EBFF><b>Benefits vs Challenges</b></font>
 
<br><br>
http://openwetware.org/wiki/IGEM:IMPERIAL/2008/Prototype/Project
{| cellpadding="5" style="background:#66aadd; border: 4px solid #66aadd; color:black" align="center" width=95%
 
!width="150px" |
Authors: Chris, James
!width="50%"|
 
<font size=4px color=white><center>'''Benefits'''</center></font>
Editors: Wiki team
!width="50%"|<font size=4px color=white><center>'''Challenges'''</center></font>
 
'''James''' - Here is a table of potential advantages and disadvatages of using ''B.subtilis'' for our project and some more general chassis ad/disad. We may wish to think about chassis ad/dis seperatly, for example, in the registry the we will want to put the chassis info up!
 
{| width="70%" cellpadding="5" style="background:#66aadd; border: 4px solid #66aadd; color:black" align="center"
|style="background:#99BBFF" width=50%|<center>'''Advantages'''</center>
|style="background:#99BBFF" width=50% width=50%|<center>'''Disadvantages'''</center>
|-
|-
|style="background:#FFFFFF"| '''Single Membrane''' - ''B.subtilis'' is a gram-positive bacterium and contains only a single membrane. This makes ''B.subtilis'' an ideal organism for secretion of proteins and other biomaterials.
|style="background:#99BBFF"|
|style="background:#FFFFFF"| '''Starting from scratch'''
|style="background:#99BBFF" colspan="2"|<font size=3px color=white><center>'''Protein Expression'''</center></font>
|-
|-
|style="background:#FFFFFF"| '''Available Research''' - Light sensing, clutch and secretion pathways
|style="background:#FFFFFF"| [[Image:Secretion.PNG| 150px]]
|style="background:#FFFFFF"|'''Peptidase activity'''
|style="background:#FFFFFF"| '''Single Membrane''' - ''B. subtilis'' is a gram-positive bacterium and contains only a single membrane. This makes ''B. subtilis'' an ideal chassis for secretion of  biomaterials.
|style="background:#FFFFFF"| '''Peptidase activity''' - ''B. subtilis'' express exopeptidases and so there is a risk that our biomaterial may be cleaved up before it can be secreted.
|-
|-
|style="background:#FFFFFF"| '''Non pathogenic''' -
|style="background:#99BBFF"|
|style="background:#FFFFFF"|'''Stationary Phase and sporulation'''
|style="background:#99BBFF" colspan="2"|<font size=3px color=white><center>'''Foundational Technology'''</center></font>
|-
|-
|style="background:#FFFFFF"| '''Naturally Competent'''
|rowspan=2 style="background:#FFFFFF" |[[Image:Gene.PNG| 150px]]
|style="background:#FFFFFF"|
| rowspan=2 style="background:#FFFFFF"| '''Availability of Potential Parts''' - ''B. subtilis'' is a highly studied organism, with a fully sequenced genome. As a result, ''B. subtilis'' provides many useful potential parts.
|style="background:#FFFFFF"| '''BioBricking''' - Although there is a huge number of potential parts, these still have to be manipulated to conform to existing BioBrick standards.
|-
|-
|style="background:#FFFFFF"| Single membrane
|style="background:#FFFFFF"| '''Starting from scratch''' - Although previous iGEM teams have looked at ''B. subtilis'' as a potential chassis, there are very few working ''B. subtilis'' parts available in the Registry. As a result, we will have to design a variety of promoter, ribosome binding sites and protein coding regions.
|style="background:#FFFFFF"|
|-
|style="background:#99BBFF"|
|style="background:#99BBFF" colspan="2"|<font size=3px color=white><center>'''BioBrick Assembly'''</center></font>
|-
|style="background:#FFFFFF"|[[Image:Integration.PNG| 150px]]
|style="background:#FFFFFF"| '''Natural Competency and Integration''' - ''B. subtilis'' has been noted for its ease and efficieny of transformation. In addition, integration of exogenous DNA into the chromosome has been well studied and provides an alternative to using traditional plasmids.   
|style="background:#FFFFFF"| '''Vector Degradation''' - ''B. subtilis'' does not use all the same vectors as ''E. coli''. One reason for this is that ''B. subtilis'' often recognises vectors grown in ''E. coli'' as foreign and digests them. Vectors and shuttle vectors are thus not reliable carriers of genetic information.
|-
|style="background:#99BBFF"|
|style="background:#99BBFF" colspan="2"|<font size=3px color=white><center>'''Chassis Properties'''</center></font>
|-
| rowspan=3 style="background:#FFFFFF"| [[Image:Chassis.PNG| 150px]]
|style="background:#FFFFFF"| '''Non-pathogenicity''' - ''B. subtilis'' is a non-pathogenic organism that is commonly found in soil. As a result, ''B. subtilis'' has a biological harzardous level of 1 and offers a useful non-pathogenic chassis for synthetic biologists.
| rowspan=3 style="background:#FFFFFF"| 
|-
|style="background:#FFFFFF"|'''High Motility''' - ''B.subtilis'' is often referred to as a highly motile organism in comparison to other bacterium.
|-
|style="background:#FFFFFF"|'''Sporulation: Transport''' - Under stress conditions, ''B. subtilis'' will form spores. These spores are highly resistant versions of single cells, able to withstand extreme temperatures and pH. These spores are capable of growing into new cells once favourable growing conditions are restored. Due to the resistance of spores, there is a great potential for manipulating them for transporting ''B. subtilis'' devices and constructs.
|}
|}
<br>
<br clear="all"><hr>
 
{{Imperial/Box1||Having decided what we're doing and how we plan to do it, the next step is to get started! To see how our lab work went please start with the [[IGEM:IMPERIAL/2008/New/Wet_Lab | '''>>> Wet Lab Hub >>>''']]}}
 
{{Imperial/EndPage|Chassis_1|Wet_Lab}}
 
 
==The Good==
 
 
* Light sensing system
 
 
 
* The Clutch
 
 
 
==The Bad==
 
* Vector digestor
 
 
 
* Starting from scratch
 
 
 
* Stationary Phase and sporulation
 
 
 
* Peptidase activity
 
 
 
==The Ugly==
 
* Take a sniff.... go on, just a quick one!
 
{{Imperial/EndPage}}

Latest revision as of 07:14, 27 September 2008

<html> <style type="text/css"> .firstHeading {display: none;} </style> </html> <html> <style type="text/css"> 

   table.calendar          { margin:0; padding:2px; }

table.calendar td { margin:0; padding:1px; vertical-align:top; } table.month .heading td { padding:1px; background-color:#FFFFFF; text-align:center; font-size:120%; font-weight:bold; } table.month .dow td { text-align:center; font-size:110%; } table.month td.today { background-color:#3366FF } table.month td { 

   border:2px;
   margin:0;
   padding:0pt 1.5pt;
   font-size:8pt;
   text-align:right;
   background-color:#FFFFFF;
   }
  1. bodyContent table.month a { background:none; padding:0 }

.day-active { font-weight:bold; } .day-empty { color:black; } </style> </html>

<html><script language="JavaScript">

var timeout = 250; var closetimer = 0; var ddmenuitem = 0;

// open hidden layer function mopen(id) { // cancel close timer mcancelclosetime(); // close old layer if(ddmenuitem) ddmenuitem.style.visibility = 'hidden'; // get new layer and show it ddmenuitem = document.getElementById(id); ddmenuitem.style.visibility = 'visible'; } // close showed layer function mclose() { if(ddmenuitem) ddmenuitem.style.visibility = 'hidden'; } // go close timer function mclosetime() { closetimer = window.setTimeout(mclose, timeout); } // cancel close timer function mcancelclosetime() { if(closetimer) { window.clearTimeout(closetimer); closetimer = null; } } // close layer when click-out //document.onclick = mclose; </script> <table background="http://i59.photobucket.com/albums/g305/Timpski/ToolbarBackground.png" style="color:#ffffff;" link="#ffffff" cellpadding="0" cellspacing="1" border="0" bordercolor="#ffffff" align="center" width="100%"><tr><td colspan="6" class="wetlab"><br><br><br></td></tr> <tr><td align="center" width="10%" valign="bottom"><ul id="sddm"><a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2008/New/Home"> Home </a></ul> </td><td align="center" width="20%" valign="bottom"><ul id="sddm"><a href="#" 

       onclick="mopen('m1')" 
       onmouseover="mopen('m1')" 
       onmouseout="mclosetime()">Biofabricator Subtilis</a>
       <div id="m1" 
           onmouseover="mcancelclosetime()" 
           onmouseout="mclosetime()">
       <a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2008/New/Project">Project Specifications</a>
       <a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2008/New/Chassis_1">Why B. subtilis?</a>
       <a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2008/New/Chassis_2">B. subtilis: Benefits vs Challenges</a>
       </div></ul>

</td><td align="center" width="18%" valign="bottom"><ul id="sddm"><a href="#" 

       onclick="mopen('m2')" 
       onmouseover="mopen('m2')" 
       onmouseout="mclosetime()">Wet Lab</a>
       <div id="m2" 
           onmouseover="mcancelclosetime()" 
           onmouseout="mclosetime()">
       <a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2008/New/Wet_Lab">Wet Lab Hub</a>
       <a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2008/New/Cloning_Strategy">Cloning Strategy</a>
       <a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2008/New/Protocols">Experiments & Protocols</a>
       <a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2008/New/Major_Results">Experimental Results</a>
       <a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2008/New/BioBricks">BioBricks & Characterisation</a>
       </div></ul>

</td><td align="center" width="18%" valign="bottom"><ul id="sddm"><a href="#" 

       onclick="mopen('m3')" 
       onmouseover="mopen('m3')" 
       onmouseout="mclosetime()">Dry Lab</a>
       <div id="m3" 
           onmouseover="mcancelclosetime()" 
           onmouseout="mclosetime()">
       <a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2008/New/Dry_Lab">Dry Lab Hub</a>
       <a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2008/New/Growth_Curve">Growth Curves</a>
       <a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2008/New/Genetic_Circuit">Genetic Circuits</a>
       <a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2008/New/Motility">Motility Analysis</a>
       <a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2008/New/Appendices">Appendices - Code etc.</a>
       </div></ul>

</td><td align="center" width="17%" valign="bottom"><ul id="sddm"><a href="http://2008.igem.org/Team:Imperial_College/Notebook"> Notebook </a></ul> </td><td align="center" width="17%" valign="bottom"><ul id="sddm"><a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2008/New/Team"> Our Team </a></ul> </td></tr></table></html>

<html><style type="text/css">

div.Section { font:11pt/16pt Calibri, Verdana, Arial, Geneva, sans-serif; background-image: url(http://openwetware.org/images/a/a0/Background.PNG); background-size: 100%; background-origin: content; }

/* Text (paragraphs) */ div.Section p { font:11pt/16pt Calibri, Verdana, Arial, Geneva, sans-serif; text-align:justify; margin-top:0px; margin-left:30px; margin-right:30px; }

/* Headings */ div.Section h1 { font:22pt Calibri, Verdana, Arial, Geneva, sans-serif; text-align:left; color:#3366FF; font-weight:bold; }

/* Subheadings */ div.Section h2 { font:18pt Calibri, Verdana, Arial, Geneva, sans-serif; color:#3366FF; margin-left:5px; font-weight:bold; }

/* Subsubheadings */ div.Section h3 { font:22pt Calibri, Verdana, Arial, sans-serif; color:#E5EBFF; margin-left:10px; font-weight:bold; }

/* Subsubsubheadings */ div.Section h4 { font:22pt Calibri, Verdana, Arial, sans-serif; color:#2B48B3; margin-left:10px; font-weight:bold; }

/* Subsubsubsubheadings */ div.Section h5 { font:12pt Calibri, Verdana, Arial, sans-serif; color:#3366FF; margin-left:20px; }

/* References */ div.Section h6 { font:12pt Calibri, Verdana, Arial, sans-serif; font-weight:bold; font-style:italic; color:#3366FF; margin-left:25px; }

/* Hyperlinks */ div.Section a { }

div.Section a:hover { }

/* Tables */ div.Section td { font:11pt/16pt Calibri, Verdana, Arial, Geneva, sans-serif; text-align:justify; vertical-align:top; padding:2px 4px 2px 4px; }

/* Lists */ div.Section li { font:11pt/16pt Calibri, Verdana, Arial, Geneva, sans-serif; text-align:left; margin-top:0px; margin-left:30px; margin-right:0px; }

/* TOC stuff */ table.toc { margin-left:10px; }

table.toc li { font: 11pt/16pt Calibri, Verdana, Arial, Geneva, sans-serif; text-align: justify; margin-top: 0px; margin-left:2px; margin-right:2px; }

/* [edit] links */ span.editsection { color:#BBBBBB; font-size:10pt; font-weight:normal; font-style:normal; vertical-align:bottom; }

span.editsection a { color:#BBBBBB; font-size:10pt; font-weight:normal; font-style:normal; vertical-align:bottom; }

span.editsection a:hover { color:#3366FF; font-size:10pt; font-weight:normal; font-style:normal; vertical-align:bottom; }

/* Drop-down Menu */

  1. sddm {

margin: 0; padding: 0; z-index: 30 margin: 0; padding: 0; float: center; font: bold 12pt Calibri, Verdana, Arial, Geneva, sans-serif; border: 0px; list-style: none; }

  1. sddm a {

display: block; margin: 0px 0px 0px 0px; padding: 0 0 12px 0; color: #FFFFFF; text-align: center; text-decoration: none; }

  1. sddm a:hover {

border: 0px }

  1. sddm div {

position: absolute; visibility: hidden; margin: 0; padding: 0; background: #66aadd; border: 1px solid #66aadd } #sddm div a { position: relative; left: 0; display: block; margin: 0; padding: 5px 10px; width: auto; white-space: nowrap; text-align: left; text-decoration: none; background: #FFFFFF; color: #2875DE; font: 11pt Calibri, Verdana, Arial, Geneva, sans-serif } #sddm div a:hover { background: #66aadd; color: #FFFFFF } </style></html>



  Benefits vs Challenges

Benefits
Challenges
Protein Expression
Single Membrane - B. subtilis is a gram-positive bacterium and contains only a single membrane. This makes B. subtilis an ideal chassis for secretion of biomaterials. Peptidase activity - B. subtilis express exopeptidases and so there is a risk that our biomaterial may be cleaved up before it can be secreted.
Foundational Technology
Availability of Potential Parts - B. subtilis is a highly studied organism, with a fully sequenced genome. As a result, B. subtilis provides many useful potential parts. BioBricking - Although there is a huge number of potential parts, these still have to be manipulated to conform to existing BioBrick standards.
Starting from scratch - Although previous iGEM teams have looked at B. subtilis as a potential chassis, there are very few working B. subtilis parts available in the Registry. As a result, we will have to design a variety of promoter, ribosome binding sites and protein coding regions.
BioBrick Assembly
Natural Competency and Integration - B. subtilis has been noted for its ease and efficieny of transformation. In addition, integration of exogenous DNA into the chromosome has been well studied and provides an alternative to using traditional plasmids. Vector Degradation - B. subtilis does not use all the same vectors as E. coli. One reason for this is that B. subtilis often recognises vectors grown in E. coli as foreign and digests them. Vectors and shuttle vectors are thus not reliable carriers of genetic information.
Chassis Properties
Non-pathogenicity - B. subtilis is a non-pathogenic organism that is commonly found in soil. As a result, B. subtilis has a biological harzardous level of 1 and offers a useful non-pathogenic chassis for synthetic biologists.
High Motility - B.subtilis is often referred to as a highly motile organism in comparison to other bacterium.
Sporulation: Transport - Under stress conditions, B. subtilis will form spores. These spores are highly resistant versions of single cells, able to withstand extreme temperatures and pH. These spores are capable of growing into new cells once favourable growing conditions are restored. Due to the resistance of spores, there is a great potential for manipulating them for transporting B. subtilis devices and constructs.

Having decided what we're doing and how we plan to do it, the next step is to get started! To see how our lab work went please start with the >>> Wet Lab Hub >>>



<html><center><table style="color:#ffffff;background-color:#66aadd;" cellpadding="3" cellspacing="1" border="0" bordercolor="#ffffff" align="center"> <tr><td><ul id="sddm"></html>< Previous<html></ul> </td><td><ul id="sddm"><a href="#">Back to top</a></ul> </td><td><ul id="sddm"></html>Next ><html></ul> </td></tr></table> </center></html>

Retrieved from "https://openwetware.org/mediawiki/index.php?title=IGEM:IMPERIAL/2008/New/Chassis_2&oldid=246520"