Regulatory
HSP

Part:BBa_K338001

Designed by: 2010 Caltech iGEM Team   Group: iGEM10_Caltech   (2010-10-24)
Revision as of 02:58, 26 October 2010 by Yuehan (Talk | contribs) (Effect of Heat Shock Temperature)

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.

Effect of Heat Shock Temperature.png


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 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.

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.

HSPGFP(1).pngHSPGFP(2).png


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]


[edit]
Categories
//rnap/prokaryote/ecoli/sigma32
//direction/forward
//chassis/prokaryote/ecoli
//promoter
//regulation/positive
Parameters
negative_regulators
positive_regulators