Difference between revisions of "Part:BBa K338002"

 
 
(4 intermediate revisions by the same user not shown)
Line 1: Line 1:
 
 
__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K338002 short</partinfo>
 
<partinfo>BBa_K338002 short</partinfo>
  
Fused heat shock promoter with lacI regulated promoter. It possesses the characteristic of both 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.
 +
 
 +
Note: there is an RBS (BBa_B0034) attached to the end of this brick.
  
<!-- Add more about the biology of this part here
 
 
===Usage and Biology===
 
===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 OD<sub>600</sub>. Normalized value of each sample at t=0 was subtracted from all the values to show a relative difference.
 +
 
 +
[[Image: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.
 +
 +
[[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.
 +
 
 +
======Image of LacI Cells Grown in 0mM Glucose======
 +
 
 +
[[Image:LacI_noglu_16(FITC).jpg|400px]][[Image:LacI_noglu_17(FITC).jpg|400px]]
 +
 
 +
======Image of LacI Cells Grown in 5mM Glucose======
 +
 
 +
[[Image:LacI_glu_23(FITC).jpg|400px]][[Image:LacI_glu_22(FITC).jpg|400px]]
 +
 
 +
'''NOTE: All images were captured at 100x magnification.'''
 +
 
 +
 
 
<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K338002 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K338002 SequenceAndFeatures</partinfo>

Latest revision as of 04:00, 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.

Note: there is an RBS (BBa_B0034) attached to the end of this brick.

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

NOTE: All images were captured at 100x magnification.


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]