Difference between revisions of "Part:BBa K216007:Experience"

Latest revision as of 22:30, 27 October 2010

This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Edinburgh 2010 Experience and Improvements

We needed this protein to shift our luxAB spectrum towards the blue in order to activate our blue light sensor. Unfortunately, when we tried to transform E.coli with the version last year's team submitted to the registry we could not get positive transformants, even in the absence of luxAB. When we examined the DNA sequence we found that the DNA was incorrect and that the wrong version had been submitted to the registry. We tried our transformations again with a different version and had more success. We then created a luxAB/lumP fusion which produced blue light in the presence of decanal. The lumazine protein, the fusion construct and the fusion construct with added promoter have been added to the registry by this year's team (BBa_K322007, BBa_K322149 and BBa_K322150 respectively) along with an image to indicate the blue shift of the fusion.

Applications of BBa_K216007

Lumazine protein (LumP) of Photobacterium phosphoreum is a small protein which binds a fluorescent pteridine, 6,7-dimethyl-8-(1'-D-ribityl)lumazine, as cofactor. In P. phosphoreum, it acts to shift the wavelength of light emitted by luciferase, LuxAB, from 495 nm to a shorter wavelength, around 475 nm (Lee et al, 1991; O'Kane et al, 1990 and references cited therein). It may be partially responsible for the much higher perceived brightness of P. phosphoreum luminescence compared to other bioluminescent bacteria such as Xenorhabdus luminescens. To find out, we hope to compare bioluminescence of E. coli expressing X. luminescens luxAB (BBa_K216008) with and without lumazine protein.

User Reviews

UNIQae28f5c5f227e4a8-partinfo-00000000-QINU UNIQae28f5c5f227e4a8-partinfo-00000001-QINU

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 This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
 how you used this part and how it worked out.
+===Edinburgh 2010 Experience and Improvements===
+We needed this protein to shift our ''luxAB'' spectrum towards the blue in order to activate our blue light sensor. Unfortunately, when we tried to transform ''E.coli'' with the version last year's team submitted to the registry we could not get positive transformants, even in the absence of ''luxAB''. When we examined the DNA sequence we found that the DNA was incorrect and that the wrong version had been submitted to the registry. We tried our transformations again with a different version and had more success. We then created a ''luxAB/lumP'' fusion which produced blue light in the presence of decanal. The lumazine protein, the fusion construct and the fusion construct with added promoter have been added to the registry by this year's team (BBa_K322007, BBa_K322149 and BBa_K322150 respectively) along with an image to indicate the blue shift of the fusion.
 ===Applications of BBa_K216007===
-Lumazine protein (LumP) of ''Photobacterium phosphoreum'' is a small protein which binds a fluorescent pteridine, 6,7-dimethyl-8-(1'-D-ribityl)lumazine, as cofactor. In ''P. phosphoreum'', it acts to shift the wavelength of light emitted by luciferase, LuxAB, from 495 nm to a shorter wavelength, around 475 nm (O'Kane et al, 1990 and references cited therein). It may be partially responsible for the much higher perceived brightness of ''P. phosphoreum'' luminescence compared to other bioluminescent bacteria such as ''Xenorhabdus luminescens''. To find out, we hope to compare bioluminescence of ''E. coli'' expressing ''X. luminescens luxAB'' (BBa_K216008) with and without lumazine protein. At the time of writing, days before the freezing of the iGEM 2009 information for judging, we have not yet been able to perform this exciting experiment, but we hope to post the results here in the near future (possibly shortly after the Jamboree).
+Lumazine protein (LumP) of ''Photobacterium phosphoreum'' is a small protein which binds a fluorescent pteridine, 6,7-dimethyl-8-(1'-D-ribityl)lumazine, as cofactor. In ''P. phosphoreum'', it acts to shift the wavelength of light emitted by luciferase, LuxAB, from 495 nm to a shorter wavelength, around 475 nm (Lee et al, 1991; O'Kane et al, 1990 and references cited therein). It may be partially responsible for the much higher perceived brightness of ''P. phosphoreum'' luminescence compared to other bioluminescent bacteria such as ''Xenorhabdus luminescens''. To find out, we hope to compare bioluminescence of ''E. coli'' expressing ''X. luminescens luxAB'' (BBa_K216008) with and without lumazine protein.
 ===User Reviews===