Measurement

Part:BBa_K911003:Experience

Designed by: Oliver Meacock   Group: iGEM12_Cambridge   (2012-09-12)
Revision as of 22:02, 25 September 2012 by Olijme (Talk | contribs) (→‎Applications of BBa_K911003)

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Applications of BBa_K911003

This riboswitch came to us from the Yale team in the following construct:

F-riboswitch.png

The lysine promotor is constitutively active in both e.coli and bacillus, so expression of β-galactosidase should only be affected by the rate of transcriptional attenuation by the fluoride riboswitch.

To verify that this part worked before undertaking more complex assays, we decided to run an assay with X-Gal to give us some qualititative data about its function. The results of this assay are shown below:

FluorideXgal.jpg

The numbers on the tops of the tubes represent the concentration (in mM) of NaF in each culture. The amount of blue is proportional to the amount of β-galactosidase activity present in the culture, which in turn is regulated by the riboswitch. This qualatative assay demonstrated that our part worked.

To get more quantatative data, we performed a Miller assay.

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