Difference between revisions of "Part:BBa K338001"
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[[Image:T0_HSP-GFP_4(FITC).jpg|400px]][[Image:T0_HSP-GFP_6(FITC).jpg|400px]] | [[Image:T0_HSP-GFP_4(FITC).jpg|400px]][[Image:T0_HSP-GFP_6(FITC).jpg|400px]] | ||
| − | Control: cells containing only GFP | + | Control: cells containing only GFP at t=0 |
[[Image:T0_GFP_12(FITC).jpg|400px]] | [[Image:T0_GFP_12(FITC).jpg|400px]] | ||
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[[Image:T10_HSP-GFP_27(FITC).jpg|400px]][[Image:T10_HSP-GFP_29(FITC).jpg|400px]] | [[Image:T10_HSP-GFP_27(FITC).jpg|400px]][[Image:T10_HSP-GFP_29(FITC).jpg|400px]] | ||
| + | |||
| + | Control: cells containing only GFP at t=10 | ||
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| + | [[Image:T10_GFP_36(FITC).jpg|400px]] | ||
======Images of Cells at t=60 min====== | ======Images of Cells at t=60 min====== | ||
[[Image:T1hr_HSP-GFP_39(FITC).jpg|400px]][[Image:T1hr_HSP-GFP_40(FITC).jpg|400px]] | [[Image:T1hr_HSP-GFP_39(FITC).jpg|400px]][[Image:T1hr_HSP-GFP_40(FITC).jpg|400px]] | ||
| + | |||
| + | Control: cells containing only GFP at t=60 | ||
| + | |||
| + | [[Image:T1hr_GFP_47(FITC).jpg|400px]] | ||
=====Glucose===== | =====Glucose===== | ||
Revision as of 03:16, 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 at t=0 min (Before Heat Shock)
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Control: cells containing only GFP at t=0
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Images of Cells at t=10 min
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Control: cells containing only GFP at t=10
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Images of Cells at t=60 min
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Control: cells containing only GFP at t=60
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Glucose
An interesting observation was made: when adding 20 mM of Glucose to the media at the time of growth,the cells are no longer morbidly elongated at the time of imaging.
Images of Cells at t=0 min (Before Heat Shock)
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Images of Cells at t=10 min
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Images of Cells at t=60 min
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Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]