Reporter
GFP genera

Part:BBa_E0840:Experience

Designed by: Jennifer Braff   Group: Antiquity   (2004-10-18)
Revision as of 02:29, 23 September 2009 by Calum (Talk | contribs)

This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_E0840

BBa_E0840 is commonly used to quantify the function of transcriptional control devices.

User Reviews

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Smelissali

Very visible even without UV excitation on plate and in solution after 24 hours incubation. Not as visible as Part:BBa_I13521 (mRFP), however.

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Reshma Shetty

BBa_E0840 was successfully used to quantify the behavior of BBa_R0040, BBa_J45992 and BBa_J45994.

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Aberdeen_Scotland 2009

Our miniprep, double and single digests worked as expected. We used this biobrick for building two of our parts (BBa_K182100 and 'BBa_K182101). We experienced very satisfactory cloning with high efficiency of transformants. The sequencing was correct. We discovered that all the bacteria containing this biobrick were releasing fluorescence even though there was no promoter upstream. The level of fluorescence observed was dependent on the plasmid copy number. We noticed that colonies displayed a visible, yellow-green colour, therefore selection was very easy.

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Characterization

Transcriptional control of GFP generator

All three transcriptional control devices were quantified using 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.