Inverter
tetR inver
Part:BBa_Q04401:Experience
Designed by: Josh Michener Group: iGEM04_MIT (2006-05-11)
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how you used this part and how it worked out.
Applications of BBa_Q04401
- Used successfully as a transcriptional inverter in the exponential phase GFP generator (BBa_J45996). See results below.
User Reviews
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In general, I think it's more likely to work when used on a high-copy (1 series) plasmid. I've tried it on a 3 series, using BBa_F2621as the induction system and did not see flipping of the inverter. |
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BBa_Q04401 worked well in the exponential phase GFP generator (BBa_J45996). |
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Characterization
Transfer curve
Transfer curve for BBa_Q04401 measured in the Screening Plasmid (pSB1A10).
Units are arbitrary. The image in top right is a dot plot based on concatenated flow cytometry data from 6 different arabinose induction levels (see pSB1A10for details). Main figure is the same dot plot data averaged to make the transfer curve more clear.
Units are arbitrary. The image in top right is a dot plot based on concatenated flow cytometry data from 6 different arabinose induction levels (see pSB1A10for details). Main figure is the same dot plot data averaged to make the transfer curve more clear.
Use in BBa_J45996: an exponential phase GFP generator composed of BBa_J45992, BBa_Q04401 and BBa_E0840
Growth phase dependent transcriptional control devices
We successfully designed, constructed and tested transcriptional control devices for constitutive, stationary phase dependent and exponential phase dependent protein production (A-C). To test and verify function of our three transcriptional control devices, we assembled each control device with the GFP protein generator BBa_E0840 and monitored the fluorescence of E. coli cultures with each device over time. For each device, we plot the change in fluorescence per unit time (normalized GFP synthesis rate) versus the cell density (OD600nm) (D). The constitutive transcriptional control device produced a high GFP synthesis rate irrespective of cell density. The stationary phase transcriptional control device produced a low initial GFP synthesis rate which increased with culture cell density. The exponential phase transcriptional control device produced an initially high GFP synthesis rate which dropped off as cell density increased. Data shown are averages of triplicate measurements of cultures grown from three individual colonies of each device. Error bars are the standard deviation of the three individual cultures.
We successfully designed, constructed and tested transcriptional control devices for constitutive, stationary phase dependent and exponential phase dependent protein production (A-C). To test and verify function of our three transcriptional control devices, we assembled each control device with the GFP protein generator BBa_E0840 and monitored the fluorescence of E. coli cultures with each device over time. For each device, we plot the change in fluorescence per unit time (normalized GFP synthesis rate) versus the cell density (OD600nm) (D). The constitutive transcriptional control device produced a high GFP synthesis rate irrespective of cell density. The stationary phase transcriptional control device produced a low initial GFP synthesis rate which increased with culture cell density. The exponential phase transcriptional control device produced an initially high GFP synthesis rate which dropped off as cell density increased. Data shown are averages of triplicate measurements of cultures grown from three individual colonies of each device. Error bars are the standard deviation of the three individual cultures.