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Team:Edinburgh/Bacterial/Blue light producer
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Revision as of 21:51, 24 October 2010
Overview: The blue light producer
The construction of a blue light producer requires the combination of luxAB with an extra protein called lumazine (lumP). This will shift the wavelength of the bacterial luciferase towards the blue spectrum, giving us a light which can activate our blue light sensor. Last year's Edinburgh team already made both of these parts so all we had to do was fuse them together and add a constitutive promoter before determining whether the wavelength of the light output had been altered.
Strategy
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Problems
The luxAB which last year's team made and stored is failing to transform correctly. Colonies that ought to be growing white are growing blue (luxAB should have replaced the lacZ' in the vector, EdinbrickI). A similar thing is happening with lumP and RFP, where cells are still growing red, despite the fact that the RFP ought to have been removed when lumP was inserted last year.
We have come across several PCR products not yet inserted into vectors with which we are hoping to start this part of the project (i.e. the bit which should already have been done by last year according to their lab notes) again.
We eventually resolved these problems by using the alternative PCR products which were perfectly capable of being cloned into pSB1C3.
Biobricks
Both lumP and luxAB have already been submitted as biobricks in pSB1A3, but if they give everyone else as much trouble as they've given us, we may have to resubmit them.
We are also submitting a luxAB-lumP fusion construct in pSB1C3 with a constitutive promoter which can be used for blue light production. We have confirmation of working luxAB with the promoter in question. Characterisation details are to come.
References
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