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|colspan="2" style="background-color: #F2F2F2;" align="right"|[[/Entry_Base|Customize your entry pages]] [[Help:Notebook/Project_Base/Customize_entry_page|<html><img src="/images/a/aa/Help.png" border="0" /></html>]]
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=Aim=
To create a system which responds to ligand binding with a detectable voltage caused by a K<sup>+</sup> flux.


[[IGEM:Cambridge/2008/Notebook/Voltage/Progress | Progress]]
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=Background=
  <tr>
[[Media:Voltage_project.ppt | Presentation]]
    <td><a href="http://www.medical-solutions.co.uk/"><img src="http://www.gen.cam.ac.uk/Images/logos/iGEMsponsors/geneservice.jpg" alt="geneservice" width="280" height="53"></a></td>
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=Experiments=
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[[IGEM:Cambridge/2008/Notebook/Voltage/Mutant Strains | Mutant Strains]]
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[[IGEM:Cambridge/2008/Notebook/Voltage/Flame Photometer Calibration|Flame Photometer Calibration]]
    <td><a href="http://www.fisher.co.uk/"><img src="http://www.gen.cam.ac.uk/Images/logos/iGEMsponsors/fisherscientific.jpg" alt="Fisher Scientific" width="265" height="66" /></a></td>
 
    <td>&nbsp;</td>
[[IGEM:Cambridge/2008/Notebook/Voltage/K+ Concentrations|K+ Concentrations]]
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[[IGEM:Cambridge/2008/Notebook/Voltage/BioBrick Manipulation|BioBrick Manipulation]]
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[[IGEM:Cambridge/2008/Notebook/Voltage/OD600 Calibration|OD600 Calibration]]
 
=Next Steps=
 
<u>Characterise promoters</u>
 
1. Simulate and design    ‘reporter plasmids’ with correct biobricks restriction enzyme sites
 
2. For each strain:    2 plasmid backbones
 
::* PSC101 (A) with    OsmY (promoter) + RFP + Stop
 
::* Another (B), not    yet defined, with Bba_J23100 + YFP + Stop
 
: Tests for each strain :  plasmid A, plasmid B, plasmid A + plasmid B (so 15 tests)
 
3. Quantify transformation    efficiency (colony counter)
 
4. Quantify promoter    strength (light intensity, expression levels)
 
=Useful Links=
 
[http://expasy.org/tools/ Protein prediction tools]
 
[http://www.uniprot.org/ Uniprot database]
 
=Literature=
 
[http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=1214631 Kdp operon diagram]
 
[http://www.jbc.org/cgi/content/abstract/276/13/9590 Kdp plasmid]
 
[http://www.springerlink.com/content/6042632827845551/ The Kdp-ATPase system and its regulation]
 
Potential Chassis: [http://cgsc.biology.yale.edu/Strain.php?ID=107402 Strain JW1242-1]
[http://cgsc.biology.yale.edu/Strain.php?ID=107065 Strain JW0710-1]
 
[http://www.ncbi.nlm.nih.gov/pubmed/4942756 Kdp mutant - paper from 1971]
 
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[http://www.shigen.nig.ac.jp/WGR/link/link_E.coli_e.html Worldwide E.coli Databases]
 
[http://jb.asm.org/cgi/content/abstract/188/5/1950 Characterisation of kdpD - 2005]
 
[http://jb.asm.org/cgi/content/full/180/19/5102 Investigations on Kdp Operon exp. & flux]
 
[http://dx.doi.org/10.1006/jmbi.2001.4884 Very interesting 2001 paper concerning Glutamate Channels]
 
[http://www.nature.com/nature/journal/v402/n6763/full/402817a0.html 1999 paper on functional characterization of prokaryote Glu Channels]
 
[http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=BA000022 Sequenced Synechocystis PCC 6803 genome]
 
[http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?val=47118304&from=1401809&to=1403002&view=gbwithparts Glutamate-gated K+ channel GluR0]
 
[http://redpoll.pharmacy.ualberta.ca/CCDB/cgi-bin/STAT_NEW.cgi Link to E.coli statistics page (CCDB Database)]
 
 
|}

Revision as of 08:14, 1 September 2008


<html> <td class="tdboxes"><a href="http://openwetware.org/wiki/iGEM:Cambridge/2008/Notebook/Voltage"><img border="0" src="http://openwetware.org/images/0/00/Vol_logo.JPG" width="256" height="201"></a></td> <td class="tdboxes"><a href="http://openwetware.org/wiki/iGEM:Cambridge/2008/Notebook/Voltage"><img border="0" src="http://openwetware.org/images/d/d1/Mag_logo.JPG" width="210" height="201"></a></td> <td class="tdboxes"><a href="http://openwetware.org/wiki/iGEM:Cambridge/2008/Notebook/Voltage"><img border="0" src="http://openwetware.org/images/f/fe/Tur_logo.JPG" width="201" height="201"></a></td> </html>


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