Team:UMaryland/project/futureapplications

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<p><img src="http://parts.igem.org/wiki/images/9/99/UMSIgnal_Cascade.jpg" border="0" width="671" height="503" style="line-height: 12.0000009536743px; display: block; margin-left: auto; margin-right: auto; cursor: se-resize !important;" /></p>
<p>One of the strategies we hope to pursue is developing or incorporating a downstream signaling cascade into the cell strain. In this effort, once the E. coli came in contact with the Dermo it would then set off a transcription factor, ultimately producing a signal.  Multiple approaches have been researched, including a Cpx membrane stress cascade, a split GFP cascade, and others.</p>
<p>One of the strategies we hope to pursue is developing or incorporating a downstream signaling cascade into the cell strain. In this effort, once the E. coli came in contact with the Dermo it would then set off a transcription factor, ultimately producing a signal.  Multiple approaches have been researched, including a Cpx membrane stress cascade, a split GFP cascade, and others.</p>
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<p>The Cpx two component system reacts naturally to membrane stress. When the much larger Dermo is bound to the surface receptor expressed on E. coli, the bacterial cell membrane would become deformed. In turn, this would transmit a signal to the nucleus via the Cpx system that could be coupled to GFP production. This assay would require florescence microscopy to detect Dermo in solution.</p>
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<p>The Cpx two component system reacts naturally to membrane stress. When the much larger Dermo is bound to the surface receptor expressed on E. coli, the bacterial cell membrane would become deformed. In turn, this would transmit a signal to the nucleus via the NlpE system that could be coupled to GFP production. This assay would require florescence microscopy to detect Dermo in solution.</p>
<p>Another option is for a split GFP construct.  The idea is two different constructs containing the ligand are present in the cell, each with a different fraction of the GFP gene. When Dermo comes in close contact with the cell, multiple ligands on the cell would bind to the receptors on Dermo, causing the membrane to pinch or come together. This would trigger a spontaneous chance that two complementary pieces of GFP in close contact would bind, producing a fluorescent signal.  This would also need to be observed using microscopy.</p>
<p>Another option is for a split GFP construct.  The idea is two different constructs containing the ligand are present in the cell, each with a different fraction of the GFP gene. When Dermo comes in close contact with the cell, multiple ligands on the cell would bind to the receptors on Dermo, causing the membrane to pinch or come together. This would trigger a spontaneous chance that two complementary pieces of GFP in close contact would bind, producing a fluorescent signal.  This would also need to be observed using microscopy.</p>
<p>One of the ultimate goals from using the downstream signaling approach is to develop a “Kamikaze Bacteria”. By this we hope that the bacteria would respond to the presence of Dermo by eliminating it, possibly through killing itself. Similar work has previously been done with bees and colony collapse.</p>
<p>One of the ultimate goals from using the downstream signaling approach is to develop a “Kamikaze Bacteria”. By this we hope that the bacteria would respond to the presence of Dermo by eliminating it, possibly through killing itself. Similar work has previously been done with bees and colony collapse.</p>

Revision as of 19:30, 17 October 2014

future applications

About Umaryland

UMaryland2014 is University of Maryland, College Parks, inaugural iGEM team. We are a combined effort of several departments and numerous faculty mentors. Although it is only our first year, believe our hard work and dedication has paid off. We can't wait for this years competition! GO TERPS!