Difference between revisions of "Part:BBa K338002"

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.

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

Image of LacI Cells Grown in 5mM Glucose

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]

@@ Line 2: / Line 2: @@
 <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.
 ===Usage and Biology===
@@ Line 8: / Line 10: @@
 ====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.
+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 suggested that heat shock combined with the addition of IPTG creates the highest level of activation.
+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.
+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.
+======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.'''
-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.
 <span class='h3bb'>Sequence and Features</span>