Biomod/2013/StJohns/results
<html>
<head>
<link href='http://fonts.googleapis.com/css?family=Open+Sans' rel='stylesheet' type='text/css'>
</head>
<style>
body {
font-family: 'Trebuchet MS', 'Open Sans', sans-serif; overflow-y: auto;
}
.container {
background-color: #ffffff; margin-top:0px
} .OWWNBcpCurrentDateFilled { display: none; }
h5 {
font-family: 'Trebuchet MS', 'Open Sans', sans-serif; font-size: 11px; font-style: normal; text-align: center; margin:0px; padding:0px;
}
- column-content
{
width: 0px; float: left; margin: 0 0 0 0; padding: 0;
} .firstHeading {
display:none; width:0px;
}
- column-one
{
display:none; width:0px; background-color: #ffffff;
}
- globalWrapper
{
width: 1024px; background-color: #ffffff; margin-left: auto; margin-right: auto
}
- content
{
margin: 0 0 0 0; align: center; padding: 12px 12px 12px 12px; width: 1000px; background-color: #ffffff; border: 0;
}
- bodyContent
{
width: 974px; align: justify; background-color: #fffffff;
}
- column-content
{
width: 1024px; background-color: #ffffff;
}
- footer
{
position: absolute; bottom: 0; width: 1024px;
}
- menu
{
position: fixed; float: left; width: 180px; padding: 10px; background-color: #FFFFFF;
}
- resolutionwarning
{
display: none; text-align: center;
}
- pagecontent
{
text-align: justify; float: right; width: 744px; margin-left: 300px; min-height: 400px
}
- toc { display: none; }
@media all {
body { background: #ffffff 0 0 no-repeat;}
}
/*Expanding list*/ ul { list-style: none; }
- exp li ul { display: none; }
- exp li:hover ul { display: block; }
- exp li a:active ul { display: block; }
a:link {color:#007788;} /*unvisited link*/ a:visited {color:#007788;} /* visited link */ a:hover {color:#00ccff;} /* mouse over link */ a:active {color:#00ccff;} /* selected link */
</style> </html>
<html><a href="http://lukemanlab.org"><img src="http://openwetware.org/images/thumb/f/f9/Lukemanlab-toehold-conga-nanny-header.jpg/180px-Lukemanlab-toehold-conga-nanny-header.jpg"></a></html>
Summary
- We have synthesized versions of the claw with (‘sticky’) and without (‘blunt’) single-stranded binding elements.
- We have synthesized versions of the claw with and without fluorescent tags for FRET analysis.
- We have visualized the above versions of the claw on AFM to show that they take the predicted shape.
- We have shown that the above versions of the claw form tight bands on a gel, indicating a single primary product of the anneal.
- We have verified that FRET-tagged origami can be visualized in a gel, providing easy visual discrimination between differently-tagged versions of the claw.
- We have demonstrated a binding interaction between the functionalized claw and functionalized capsid as well as a lack of interaction between the nonfunctionalized claw and capsid.
- We have not been able to differentiate bound and unbound complexes via DLS.
- We have generated and isolated FAB fragments for use as binding elements in future claw designs.
- We have demonstrated the potential for selecting claws based on their avidity using a chromatography augmented with photocleavable elements.
Data
Claw Visualization
Below are AFM images of the claws alone, with and without binding elements. <html><center><table><tbody align="center"><tr><td><img src="http://openwetware.org/images/thumb/6/66/Lukemanlab-Bluntclaw_afm.png/200px-Lukemanlab-Bluntclaw_afm.png"></td><td><img src="http://openwetware.org/images/thumb/9/9f/Lukemanlab-Stickyclaw_afm.png/200px-Lukemanlab-Stickyclaw_afm.png"></td></tr><tr><td>"Blunt" claw</td><td>"Sticky" claw</td></tr></tbody></table></center></html>
Binding Interaction
We used gel electrophoresis to characterize the binding interaction between origami structures and capsids with and without binding elements.
‘sticky’ DO bind sticky capsids strongly;
however, blunt DO binds sticky capsids.
FRET
Below is a gel demonstrating that versions of the claw tagged with different fluorescent elements can be visually distinguished on a gel.
The six bands on the left are 'blunt' claw and the six on the right are 'sticky' claw.
Odd-numbered lanes contain 0.1 pmol claw, while even-numbered lanes contain 0.2 pmol.
