Composite

Part:BBa_K098995:Experience

Designed by: Harvard iGEM 08   Group: iGEM08_Harvard   (2008-10-26)
Revision as of 11:06, 26 July 2011 by Tokn (Talk | contribs)

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Applications of BBa_K098995

User Reviews

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BBa_K098995 Kun Xue

In order to characterize the thermoinducible promoter properly, we have used 3 sets of control cells (not shown in graph):

  • Positive control cells: Containing the BBa_I13522, acting as a baseline comparison by constitutively expressing GFP.
  • Negative control cells: These contain the thermoinducible promoter on its own ( BBa_K098995) with no GFP attached to it. These serve as cells without any fluorescence.
  • Cells shifted from 28 to 42 degrees. This characterises the transition stage.


II09 Thermoconstruct main2.jpg
The characterization of the thermoinducible promoter is as follows:

  • At 28 ºC, there is no fluorescence (GFP) output because its expression is repressed
  • At 42 ºC, we can observe an increase in fluorescence (GFP) output,as its expression is no longer repressed

Hence, we can conclude that an increase in temperature de-represses the expression of the thermoinducible system.

As part of Imperial iGEM 2009 [http://2009.igem.org/Team:Imperial_College_London| The E.ncapsulator] project, we characterised this part using the composite BBa_K200022.



The 2011 iGEM Team of Freiburg tried to use this part. Sequencing showed several inconsistencies compared to the submitted sequence. Especially between the RBS and the start codon of the repressor 6 bases are missing, probably leading to a diminished expression of the repressor.

Freigem added the missing nts between RBS and cI by QuickChange PCR. The functionality of the corrected part is currently under investigation. The new annotated sequence can be downloaded [http://2011.igem.org/Team:Freiburg| not here].


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