BBa K338002

From 2010.igem.org


iGEM 2010



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K338001+R0011: Heat Shock Promoter + LacI Regulated Promoter

Fused heat shock promoter with lacI regulated promoter. It possesses the characteristics of both a heat shock promoter and LacI.

Note: there is an RBS (BBa_B0034) attached to the end of this brick.

Usage and Biology

BioBrick Characterization

Effect of Heat Shock Temperature

A DH5α cell culture containing HSP-LacI-GFP was divided into four samples. At the beginning of the incubation period, IPTG was added to Samples 2 and 3. Then Samples 3 and 4 were heat shocked at 42°C for 2 hours. At the end of the incubation period, measurements of the fluorescence level of GFP were taken using the plate reader at 20 minute intervals and were averaged over a 1 hour interval. The vertical axis is raw fluorescence units normalized by OD600. Normalized value of each sample at t=0 was subtracted from all the values to show a relative difference.

Effect of IPTG and Heat Shock Duraton.png

Experiment suggests that heat shock combined with IPTG creates the highest level of activation.

Effect of Glucose

DH5α cells containing HSP-LacI-GFP were grown in LB solution with 0 mM and 5 mM glucose with a controlled culture containing only HSP. Measurements of fluorescence level of GFP were taken using the plate reader.

Effect of Glucose.png

In non-LacIq cells such as DH5α, LacI is not fully suppressed. Glucose was used to regulate the LacI promoter. As shown above, with the addition of 5 mM glucose, the production of GFP decreased 5 fold compared to cells grown in glucose-free solution.

Image of LacI Cells Grown in 0mM Glucose

LacI noglu 16(FITC).jpgLacI noglu 17(FITC).jpg

Image of LacI Cells Grown in 5mM Glucose

LacI glu 23(FITC).jpgLacI glu 22(FITC).jpg

NOTE: All images were captured at 100x magnification.

 
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