Difference between revisions of "IGEM:Cambridge/2008/Notebook/Voltage"

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Revision as of 14:29, 19 August 2008



<!-- sibboleth --><div id="lncal1" style="border:0px;"><div style="display:none;" id="id">lncal1</div><div style="display:none;" id="dtext">07/15/2008,07/16/2008,07/17/2008,07/18/2008,07/21/2008,07/22/2008,07/23/2008,07/24/2008,07/25/2008,07/28/2008,07/29/2008,07/30/2008,07/31/2008,08/01/2008,08/04/2008,08/05/2008,08/06/2008,08/07/2008,08/08/2008,08/11/2008,08/12/2008,08/13/2008,08/14/2008,08/15/2008,08/18/2008,08/19/2008,08/20/2008,08/21/2008,08/22/2008,08/25/2008,08/26/2008,08/27/2008,08/28/2008,08/29/2008,09/02/2008,09/03/2008,09/04/2008,09/05/2008,09/08/2008,09/09/2008,09/10/2008,09/11/2008,09/12/2008,09/15/2008,09/16/2008,09/17/2008,09/18/2008,09/19/2008,09/22/2008,09/23/2008,09/24/2008,09/25/2008,09/26/2008,09/29/2008,09/30/2008</div><div style="display:none;" id="page">IGEM:Cambridge/2008/Notebook/Voltage</div><div style="display:none;" id="fmt">yyyy/MM/dd</div><div style="display:none;" id="css">OWWNB</div><div style="display:none;" id="month"></div><div style="display:none;" id="year"></div><div style="display:none;" id="readonly">Y</div></div>

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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

Kdp 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

16 October 2017

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