Difference between revisions of "Part:BBa K338001"

Revision as of 23:54, 24 October 2010

Heat Shock Promoter (HSP)

This is a modified K112400 heat shock promoter in BBa standard.

Usage and Biology

BioBrick Characterization

Effect of Heat Shock Temperature

DH5α cells containing HSP-GFP were incubated at 4°C, 25°C, 42°C, and 45°C separately for two hours. 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 4°C at t=0 was subtracted from all the values to show a relative difference.

This experiment data suggested that heat shock promoter activity increased with heat shock temperature

Effect of Heat Shock Duration

DH5α cells containing HSP-GFP were heat shocked at 42°C for 0 min (kept at room temperature), 5 min and 10 min separately. Measurements of fluorescence level of GFP were taken using the plate reader at 15 minutes interval and were averaged over 1 hour interval. The vertical axis is raw fluorescence units normalized by OD₆₀₀.

The measurements suggest that a heat shock period of 10 minutes significantly increase the production of GFP, while a heat shock period of 5 minutes leads to a similar GFP production level as the not heat shocked cells. Furthermore, the production of GFP shows a decreasing trend after 3 hours suggesting that the heat shock promoter was only activated for a certain time period. DH5α Cells contained in the images below were incubated at 42°C for 10 minutes, and 6 hours later the fluorescence visualized by an optical microscope with a GFP-specific filter.

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 1: / Line 1: @@
 __NOTOC__
 <partinfo>BBa_K338001 short</partinfo>
@@ Line 5: / Line 4: @@
 This is a modified K112400 heat shock promoter in BBa standard.
-<!-- Add more about the biology of this part here
 ===Usage and Biology===
-<!-- -->
+====BioBrick Characterization====
+=====Effect of Heat Shock Temperature=====
+DH5α cells containing HSP-GFP were incubated at 4°C, 25°C, 42°C, and 45°C separately for two hours. 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 4°C at t=0 was subtracted from all the values to show a relative difference.
+[[Image:Effect of Heat Shock Temperature.png]]
+This experiment data suggested that heat shock promoter activity increased with heat shock temperature
+=====Effect of Heat Shock Duration=====
+DH5α cells containing HSP-GFP were heat shocked at 42°C for 0 min (kept at room temperature), 5 min and 10 min separately. Measurements of fluorescence level of GFP were taken using the plate reader at 15 minutes interval and were averaged over 1 hour interval. The vertical axis is raw fluorescence units normalized by OD<sub>600</sub>.
+[[Image:Effect of Heat Shock Pulse Duration.png]]
+The measurements suggest that a heat shock period of 10 minutes significantly increase the production of GFP, while a heat shock period of 5 minutes leads to  a similar GFP production level as the not heat shocked cells. Furthermore, the production of GFP shows a decreasing trend after 3 hours suggesting that the heat shock promoter was only activated for a certain time period.
+DH5α Cells contained in the images below were incubated at 42°C for 10 minutes, and 6 hours later the fluorescence visualized by an optical microscope with a GFP-specific filter.
+[[Image:HSPGFP(1).png]][[Image:HSPGFP(2).png]]
 <span class='h3bb'>Sequence and Features</span>
 <partinfo>BBa_K338001 SequenceAndFeatures</partinfo>