Difference between revisions of "Part:BBa K338002"
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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. | 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. | ||
| − | [[Image:Effect of Glucose.png]] | + | [[Image:Effect of Glucose.png|400px]] |
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. | 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. | ||
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| + | ======Image of LacI Cells Grown in 0mM Glucose====== | ||
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| + | [[Image:LacI_noglu_16(FITC).jpg|400px]][[Image:LacI_noglu_17(FITC).jpg|400px]] | ||
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| + | ======Image of LacI Cells Grown in 5mM Glucose====== | ||
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| + | [[Image:LacI_glu_23(FITC).jpg|400px]][[Image:LacI_glu_22(FITC).jpg|400px]] | ||
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<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> | ||
Revision as of 03:25, 27 October 2010
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.
Image of LacI Cells Grown in 0mM Glucose
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Image of LacI Cells Grown in 5mM Glucose
.jpg)
.jpg)
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]