Composite

Part:BBa_K200022:Experience

Designed by: Royah Vaezi   Group: iGEM09_Imperial College London   (2009-10-15)

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Applications of BBa_K200022

User Reviews

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BBa_K200022 Review 1 Not understood Kun

Cells were grown at the 2 different temperatures of 28°C and 42°C. There was in addition a set of cells that were shifted from 28°C to 42°C so that we could characterise the change in GFP fluorescence during the transition between temperatures.

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.

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