Part:BBa_K338002
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.
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.
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.
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.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
//direction/forward
//chassis/prokaryote/ecoli
//promoter
//regulation/positive
| negative_regulators | 1 |
| positive_regulators |