IGEM:Cambridge/2008/Notebook/Voltage

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=Aim=
=Aim=
To create a system which responds to ligand binding with a detectable voltage caused by a K<sup>+</sup> flux.
To create a system which responds to ligand binding with a detectable voltage caused by a K<sup>+</sup> flux.
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[[IGEM:Cambridge/2008/Notebook/Voltage/Progress |<font style="color:#f3ff1f">Progress</font>]]
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[[IGEM:Cambridge/2008/Notebook/Voltage/Progress |Progress]]
=Background=
=Background=
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[[Media:Voltage_project.ppt |<font style="color:#f3ff1f">Presentation</font>]]
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[[Media:Voltage_project.ppt |Presentation]]
=Experiments=
=Experiments=
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[[IGEM:Cambridge/2008/Notebook/Voltage/Mutant Strains |<font style="color:#f3ff1f"> Mutant Strains</font> ]]
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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|<font style="color:#f3ff1f">Flame Photometer Calibration</font>]]
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[[IGEM:Cambridge/2008/Notebook/Voltage/Flame Photometer Calibration|Flame Photometer Calibration]]
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[[IGEM:Cambridge/2008/Notebook/Voltage/K+ Concentrations|<font style="color:#f3ff1f">K+ Concentrations</font>]]
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[[IGEM:Cambridge/2008/Notebook/Voltage/K+ Concentrations|K+ Concentrations]]
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[[IGEM:Cambridge/2008/Notebook/Voltage/BioBrick Manipulation|<font style="color:#f3ff1f">BioBrick Manipulation</font>]]
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[[IGEM:Cambridge/2008/Notebook/Voltage/BioBrick Manipulation|BioBrick Manipulation]]
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[[IGEM:Cambridge/2008/Notebook/Voltage/OD600 Calibration|<font style="color:#f3ff1f">OD600 Calibration </font> ]]
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[[IGEM:Cambridge/2008/Notebook/Voltage/OD600 Calibration|OD600 Calibration]]
=Next Steps=
=Next Steps=
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=Useful Links=
=Useful Links=
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[http://expasy.org/tools/ <font style="color:#f3ff1f">Protein prediction tools</font>]
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[http://expasy.org/tools/ Protein prediction tools]
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[http://www.uniprot.org/ <font style="color:#f3ff1f">Uniprot database</font>]
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[http://www.uniprot.org/ Uniprot database]
=Literature=
=Literature=
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[http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=1214631|<font style="color:#f3ff1f"> Kdp operon diagram</font>]
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[http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=1214631| Kdp operon diagram]
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[http://www.jbc.org/cgi/content/abstract/276/13/9590|<font style="color:#f3ff1f"> Kdp plasmid</font>]
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[http://www.jbc.org/cgi/content/abstract/276/13/9590|Kdp plasmid]
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[http://www.springerlink.com/content/6042632827845551/ <font style="color:#f3ff1f"> The Kdp-ATPase system and its regulation</font>]
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[http://www.springerlink.com/content/6042632827845551/ The Kdp-ATPase system and its regulation]
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Potential Chassis: [http://cgsc.biology.yale.edu/Strain.php?ID=107402 |<font style="color:#f3ff1f">Strain JW1242-1</font>]
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Potential Chassis: [http://cgsc.biology.yale.edu/Strain.php?ID=107402 |Strain JW1242-1]
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[http://cgsc.biology.yale.edu/Strain.php?ID=107065 <font style="color:#f3ff1f">Strain JW0710-1</font>]
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[http://cgsc.biology.yale.edu/Strain.php?ID=107065 Strain JW0710-1]
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[http://www.ncbi.nlm.nih.gov/pubmed/4942756 <font style="color:#f3ff1f">Kdp mutant - paper from 1971</font>]
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[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 <font style="color:#f3ff1f"> Worldwide E.coli Databases</font>]
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[http://www.shigen.nig.ac.jp/WGR/link/link_E.coli_e.html Worldwide E.coli Databases]
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[http://jb.asm.org/cgi/content/abstract/188/5/1950 <font style="color:#f3ff1f">Characterisation of kdpD - 2005</font>]
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[http://jb.asm.org/cgi/content/abstract/188/5/1950 Characterisation of kdpD - 2005]
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[http://jb.asm.org/cgi/content/full/180/19/5102 <font style="color:#f3ff1f">Investigations on Kdp Operon exp. & flux</font>]
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[http://jb.asm.org/cgi/content/full/180/19/5102 Investigations on Kdp Operon exp. & flux]
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[http://dx.doi.org/10.1006/jmbi.2001.4884 <font style="color:#f3ff1f"> Very interesting 2001 paper concerning Glutamate Channels</font>]
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[http://dx.doi.org/10.1006/jmbi.2001.4884 Very interesting 2001 paper concerning Glutamate Channels]
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[http://www.nature.com/nature/journal/v402/n6763/full/402817a0.html <font style="color:#f3ff1f">1999 paper on functional characterization of prokaryote Glu Channels</font>]
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[http://www.nature.com/nature/journal/v402/n6763/full/402817a0.html 1999 paper on functional characterization of prokaryote Glu Channels]
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[http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=BA000022 <font style="color:#f3ff1f"> Sequenced Synechocystis PCC 6803 genome</font>]
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[http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=BA000022 Sequenced Synechocystis PCC 6803 genome]
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[http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?val=47118304&from=1401809&to=1403002&view=gbwithparts <font style="color:#f3ff1f"> Glutamate-gated K+ channel GluR0</font>]
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[http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?val=47118304&from=1401809&to=1403002&view=gbwithparts   Glutamate-gated K+ channel GluR0]
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[http://redpoll.pharmacy.ualberta.ca/CCDB/cgi-bin/STAT_NEW.cgi|<font style="color:#f3ff1f"> Link to E.coli statistics page (CCDB Database)</font>]
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[http://redpoll.pharmacy.ualberta.ca/CCDB/cgi-bin/STAT_NEW.cgi| Link to E.coli statistics page (CCDB Database)]
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Revision as of 11:46, 1 September 2008

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Aim

To create a system which responds to ligand binding with a detectable voltage caused by a K+ flux.

Progress

Background

Presentation

Experiments

Mutant Strains

Flame Photometer Calibration

K+ Concentrations

BioBrick Manipulation

OD600 Calibration

Next Steps

1. Ligate OsmY, RBS, and KDP in sequence into a vector. Possible difficulty due to the size of the KDP gene.

2. Determine the correct mutant to use:

  • Determine transformation efficiency of mutants.
  • Plot growth curves of mutants in normal/varying K+ conditions.
  • Estimate K+ uptake in varying K+ conditions.

3. Acquire ligand gated ion channel (GluR0)

Useful Links

Protein prediction tools

Uniprot database

Literature

Kdp operon diagram

plasmid

The Kdp-ATPase system and its regulation

Potential Chassis: |Strain JW1242-1 Strain JW0710-1

Kdp mutant - paper from 1971

Worldwide E.coli Databases

Characterisation of kdpD - 2005

Investigations on Kdp Operon exp. & flux

Very interesting 2001 paper concerning Glutamate Channels

1999 paper on functional characterization of prokaryote Glu Channels

Sequenced Synechocystis PCC 6803 genome

Glutamate-gated K+ channel GluR0

Link to E.coli statistics page (CCDB Database)

Recent changes



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