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Team:Cambridge/Project/Amplification/Characterisation
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== Introduction == | == Introduction == | ||
| - | The Cambridge 2007 iGEM team build 15 "amplifiers," constructs with RFP and GFP reporters that amplified the PoPS output of the promoter pBad/AraC(<partinfo>BBa_I0500</partinfo>, as described below: | + | The Cambridge 2007 iGEM team build 15 "amplifiers," constructs with RFP and GFP reporters that amplified the PoPS output of the promoter pBad/AraC(<partinfo>BBa_I0500</partinfo>), as described below: |
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== Characterisation == | == Characterisation == | ||
Revision as of 20:43, 21 October 2009
Categories :
Project :
-
Overview
Sensitivity Tuner
--- Characterisation
--- Modelling
Colour Generators
--- Carotenoids (Orange/Red)
--- Melanin (Brown)
--- Violacein (Purple/Green)
The Future
Safety
Notebook :
Team Logistics :
The Sensitivity Tuner
Introduction
The Cambridge 2007 iGEM team build 15 "amplifiers," constructs with RFP and GFP reporters that amplified the PoPS output of the promoter pBad/AraC(), as described below:
We re-designed these constructs to be PoPS converters as follows...
...and generated our own set of Sensitivity Tuners:
| P2 ogr activator | PSP3 pag activator | phiR73 delta activator | |
|---|---|---|---|
| PF promoter | |||
| PO promoter | |||
| PP promoter | |||
| Psid promoter | |||
| PLL promoter |
In order to characterize these phage activator/promoter constructs, we used the corresponding Cambridge 2007 amplifier as an illustration of how our Sensitivity Tuners alter the behaviour of pBad/AraC. These parts are very useful for characterisation as they contain fluorescent reporters; the parts we designed, which lack an input promoter and fluorescent reporters, are more useful parts for other iGEM teams to incorporate into their own projects. For characterisation, we moved the Cambridge 2007 amplifiers onto a low copy plasmid in order to make meaningful comparisons with , the standard promoter. We looked at four major characteristics relating input (arabinose) to output (GFP) and how they are modified compared to pBad/AraC on its own.
Characterisation
We moved all 15 activator constructs onto pSB3K3, a low copy plasmid. The standard promoter for 1 RPU, J69591, is also on pSB3K3 and has a GFP reporter, so we can make meaningful comparisons on the plate reader.
Maximum Rates against Arabinose Concentrations
80
81
82
84
85
90
92
94
95
Reconstruction
Below is a table listing the sensitivity tuners as they appear in the registry as biobricks:
