Difference between revisions of "Part:BBa K338001"

Revision as of 02:43, 27 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 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 4°C at t=0 was subtracted from all the values to show a relative difference.

This experiment data suggests that heat shock promoter activity increases 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 the fluorescence level of GFP were taken using the plate reader at 15 minute intervals and were averaged over a 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 increases the production of GFP, while a heat shock period of 5 minutes leads to a similar GFP production level as the untreated 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-visualized by an optical microscope with a GFP-specific filter- were incubated at 42°C for 10 minutes.

Images of Cells Before Heat Shock

and 6 hours later

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 23: / Line 23: @@
 The measurements suggest that a heat shock period of 10 minutes significantly increases the production of GFP, while a heat shock period of 5 minutes leads to a similar GFP production level as the untreated 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 was visualized by an optical microscope with a GFP-specific filter.
+DH5α cells contained in the images below-visualized by an optical microscope with a GFP-specific filter- were incubated at 42°C for 10 minutes.
+======Images of Cells Before Heat Shock======
+[[Image:T0_HSP-GFP_4(FITC).jpg]][[Image:T0_HSP-GFP_6(FITC).jpg]]
+ and 6 hours later
 [[Image:HSPGFP(1).png]][[Image:HSPGFP(2).png]]