Regulatory

Part:BBa_K875001:Experience

Designed by: Federico Colombo, Elisa Clagnan   Group: iGEM12_Trieste   (2012-09-23)
Revision as of 15:42, 24 September 2012 by Colombofederico (Talk | contribs)


Trieste Team 2012

To test this promoter we performed two different assays. First of all we cloned this promoter in pSB1C3 and then we inserted downstream the GFP BBa_I13504. In the same plasmid we cloned J23100-CymR-B0015 in order to repress the promoter. To test this system we used different concentrations of cumate which binds CymR preventing its repression, thus allowing the GFP expression.

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In liquid assay:

We inoculated a 20ml culture. After overnight growth, we diluted the culture to OD600 = 0.2. Then we aliquoted 200 μl in 8 replicates in a microtiter plate at different concentrations of cumate. The reading was performed in a monochromator at 485-510nm.

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In the plate assay:


We streaked the culture on LB Agar plates containing different p-cumate concentrations.

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