Difference between revisions of "Part:BBa K338002"

(Effect of Glucose)
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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]]
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[[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.

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


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]