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BBa K338002
From 2010.igem.org
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<partinfo>BBa_K338002 short</partinfo> | <partinfo>BBa_K338002 short</partinfo> | ||
Fused heat shock promoter with lacI regulated promoter. It possesses the characteristics of both a heat shock promoter and LacI. | Fused heat shock promoter with lacI regulated promoter. It possesses the characteristics of both a heat shock promoter and LacI. | ||
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| + | Note: there is an RBS (BBa_B0034) attached to the end of this brick. | ||
===Usage and Biology=== | ===Usage and Biology=== | ||
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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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| + | '''NOTE: All images were captured at 100x magnification.''' | ||
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Latest revision as of 04:02, 27 October 2010
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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 BiologyBioBrick CharacterizationEffect of Heat Shock TemperatureA 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 GlucoseDH5α 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
Image of LacI Cells Grown in 5mM Glucose
NOTE: All images were captured at 100x magnification.
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