Difference between revisions of "Part:BBa K091112:Experience"

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This part worked as expected in HB101 cells.<br>
 
This part worked as expected in HB101 cells.<br>
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New information provided by the MoWestern/Davidson team in 2009. See note at the bottom of the page. <br>
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[[Image:Trial1.png]]<br>
 
[[Image:Trial1.png]]<br>
 
[[Image:Trial2.png]]
 
[[Image:Trial2.png]]
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===User Reviews===
 
===User Reviews===
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The MWSU/Davdison iGEM 2009 team sequenced this promoter after there was little to no expression using this it to express GFP and after obtaining what seemed like faulty results during gel verification of its size. The team found that there was a 33 bp insertion within this promoter before the suffix. We believe that the  
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The MWSU/Davidson iGEM 2009 team sequenced this promoter isolated from cells that had grown for several weeks when we obtained gel results indicating that the part might be too large. The team found that there was a 35 bp spontaneous insertion within this promoter directly before the suffix. We believe that the ''E.coli'' cells may have mutated the promoter to silence its activity in an effort to conserve energy and select against an attribute that did not necessarily improve its fitness. Here is the 35 bp insertion:
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E.coli cells may have mutated the promoter to silence its activity in an effort to conserve energy and selection against an attribute that did not necessarily improve its fitness.
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  TGTGTGGAATTGTGAGCGGATAACAATTTCACACA
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Our experience with this part and our conclusion concerning the mutation have been added to the promoter description in the Parts Registry so that other teams will be aware of this unique possibility. Users are encouraged to sequence parts using this promoter during the construction process. We have some versions that are wild-type and others that are mutated. If you build a new version yourself, do not maintain the plasmid in cultures for very long. Freeze down a stock as soon as possible to avoid the introduction of a silencing mutation in the promoter.
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Latest revision as of 12:58, 31 October 2009

This part worked as expected in HB101 cells.
New information provided by the MoWestern/Davidson team in 2009. See note at the bottom of the page.

Trial1.png
Trial2.png


Applications of BBa_K091112

User Reviews

The MWSU/Davidson iGEM 2009 team sequenced this promoter isolated from cells that had grown for several weeks when we obtained gel results indicating that the part might be too large. The team found that there was a 35 bp spontaneous insertion within this promoter directly before the suffix. We believe that the E.coli cells may have mutated the promoter to silence its activity in an effort to conserve energy and select against an attribute that did not necessarily improve its fitness. Here is the 35 bp insertion: 

 TGTGTGGAATTGTGAGCGGATAACAATTTCACACA

Our experience with this part and our conclusion concerning the mutation have been added to the promoter description in the Parts Registry so that other teams will be aware of this unique possibility. Users are encouraged to sequence parts using this promoter during the construction process. We have some versions that are wild-type and others that are mutated. If you build a new version yourself, do not maintain the plasmid in cultures for very long. Freeze down a stock as soon as possible to avoid the introduction of a silencing mutation in the promoter.


UNIQd706eaa41f853f43-partinfo-00000000-QINU UNIQd706eaa41f853f43-partinfo-00000001-QINU