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Team:Cambridge/Project/VI03
From 2009.igem.org
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'''Successful Pigment Production''' | '''Successful Pigment Production''' | ||
| - | We transformed Top10 with pPSX-Vio+. After three colour eventually appeared, as shown below. Interestingly, the pigment appears to remain within the bacteria, with little or no bleeding into the media. We took the violacein pigment bacteria (right plate in photo) out of the fridge to find that the purple colour had started to develop. They were therefore left at room temperature overnight. The colour appears to be within the bacteria, with little or no bleeding into the media. The control plate (left plate) is the untransformed TOP10 E. coli. | + | We transformed Top10 with pPSX-Vio+. After three colour eventually appeared, as shown below. Interestingly, the pigment appears to remain within the bacteria, with little or no bleeding into the media. We took the violacein pigment bacteria (right plate in photo) out of the fridge to find that the purple colour had started to develop. They were therefore left at room temperature overnight. The colour appears to be within the bacteria, with little or no bleeding into the media. The control plate (left plate) is the untransformed TOP10 ''E. coli''. |
[[Image:Cambridge Violacein Pigment.jpg | 300px]] | [[Image:Cambridge Violacein Pigment.jpg | 300px]] | ||
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The Vio operon is currently on a very low copy number plasmid; moving it onto a higher copy number plasmid may accelerate pigment production. | The Vio operon is currently on a very low copy number plasmid; moving it onto a higher copy number plasmid may accelerate pigment production. | ||
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=== Characterisation of colour output === | === Characterisation of colour output === | ||
Revision as of 22:08, 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 :
Violacein Pigments
Characterisation
Proof of pigment production
Successful Pigment Production
We transformed Top10 with pPSX-Vio+. After three colour eventually appeared, as shown below. Interestingly, the pigment appears to remain within the bacteria, with little or no bleeding into the media. We took the violacein pigment bacteria (right plate in photo) out of the fridge to find that the purple colour had started to develop. They were therefore left at room temperature overnight. The colour appears to be within the bacteria, with little or no bleeding into the media. The control plate (left plate) is the untransformed TOP10 E. coli.
Left: control plate - untransformed TOP10 E. coli, Right: Top10 transformed with pPSX-Vio+.
Pigment production efficienty
The Vio operon is currently on a very low copy number plasmid; moving it onto a higher copy number plasmid may accelerate pigment production.
Characterisation of colour output
We characterised the violacein pigment by carrying out the acetone extraction protocol used for carotene. The results were normalised for OD and then plotted as a graph of absorption units against wavelength:
We also looked at absorbance at 584nm (the maximum absorbance for violacein):
