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)
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