IGEM:Cambridge/2008/Notebook/Voltage

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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



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