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{{Caltech_iGEM_10| Content= <br /> 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. [[Image: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 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. [[Image:Effect of Heat Shock Pulse Duration.png]] 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)====== [[Image:T0_HSP-GFP_4(FITC).jpg|400px]][[Image:T0_HSP-GFP_6(FITC).jpg|400px]] Control: cells containing only GFP at t=0 [[Image:T0_GFP_12(FITC).jpg|400px]] ======Images of Cells at t=10 min====== [[Image:T10_HSP-GFP_27(FITC).jpg|400px]][[Image:T10_HSP-GFP_29(FITC).jpg|400px]] Control: cells containing only GFP at t=10 [[Image:T10_GFP_36(FITC).jpg|400px]] ======Images of Cells at t=60 min====== [[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===== 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)====== [[Image:T0_HSP-GFP-glu_8(FITC).jpg|400px]][[Image:T0_HSP-GFP-glu_10(FITC).jpg|400px]] ======Images of Cells at t=10 min====== [[Image:T10_HSP-GFP-glu_30(FITC).jpg|400px]][[Image:T10_HSP-GFP-glu_32(FITC).jpg|400px]] ======Images of Cells at t=60 min====== [[Image:T1hr_HSP-GFP-glu_44(FITC).jpg|400px]] '''NOTE: All images were captured at 100x magnification.''' &nbsp; }}

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