Difference between revisions of "Part:BBa I7100:Experience"

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This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
 
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--[[User:Kahaynes|Kahaynes]] 15:45, 23 October 2006 (EDT)
 
--[[User:Kahaynes|Kahaynes]] 15:45, 23 October 2006 (EDT)
 
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<partinfo>BBa_I7100 AddReview 5</partinfo><br>
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<I>[[User:Rshetty|Reshma Shetty]]</I>
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<partinfo>BBa_I7100</partinfo> produced constitutive fluorescence in the absence of TetR. 
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==Characterization==
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===Transcriptional control of GFP generator===
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[Note: <partinfo>BBa_I7100</partinfo> is a composite part of <partinfo>BBa_R0040</partinfo> and <partinfo>BBa_E0840</partinfo>.]
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{{:iGEM:MIT/2006/Transcriptional control devices}}
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Latest revision as of 01:11, 8 March 2008

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_I7100

User Reviews

UNIQ4e402d1850232e54-partinfo-00000000-QINU

Antiquity

This review comes from the old result system and indicates that this part did not work in some test.

Kahaynes

This part is supposed to be in well 10K of iGEM 2006 DNA-2, but what is actually in this well is probably a completely different part. We cloned this part in E. coli strain JM109 and saw no GFP expression. The vector is supposed to be pSB3K3 (2750 bp) and the insert should be ~950 bp, but a restriction digest showed the vector to be > 3000 bp and the insert is > 1000 bp. This plasmid should convey Kan resistance only, but the clones are both Amp and Kan resistant.

The digests used to confirm the plasmids at each step looked clean (two clear bands) so the content of well 10K of iGEM 2006 DNA-2 is probably a BioBrick part, just not the right vector or part. The vector is probably pSB*AK*.

--Kahaynes 15:45, 23 October 2006 (EDT)

•••••

Reshma Shetty

BBa_I7100 produced constitutive fluorescence in the absence of TetR.

UNIQ4e402d1850232e54-partinfo-00000005-QINU

Characterization

Transcriptional control of GFP generator

[Note: BBa_I7100 is a composite part of BBa_R0040 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.