Part:BBa_K338002
K338001+R0011: Heat Shock Promoter + LacI Regulated Promoter
Fused heat shock promoter with lacI regulated promoter. It possesses the characteristic of both 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 incubation period, IPTG was added to Sample 2 and 3, then sample 3 and 4 were heat shocked at 42°C for 2 hours. At the end of the incubation period, measurements of fluorescence level of GFP were taken using the plate reader at 20 minutes interval and were averaged over 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 suggested that heat shock combined with the addition of 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 comparing to cells grown in glucose free solution decreased 5 folds.
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 |