Difference between revisions of "Part:BBa K606036:Experience"

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This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
 
how you used this part and how it worked out.
 
  
</html>
 
{| border="1" class="wikitable" style="text-align: center;"
 
|+T7 emitter RFP + Control in E.coli at 37°C
 
|-
 
|[[File:T7 autoloop ctrl-7.jpg|450px|thumb|center|E.coli  at 37°C  (trans image)]]
 
|[[File:T7 autoloop ctrl-9.jpg|450px|thumb|center|E.coli  at 37°C  (rfp image)]]
 
|-
 
|[[File:T7 autoloop ctrl-31.jpg|450px|thumb|center|E.coli  negative control at 37°C  (trans image)]]
 
|[[File:T7 autoloop ctrl-33.jpg|450px|thumb|center|E.coli  negative control at 37°C  (rfp image)]]
 
|}
 
 
 
The first pictures show that the RFP construct is working efficiently since some cells are glowing with RFP fluorescence. This also shows the system is not as leaky as we expected.  Indeed, the promoter regulating RFP expression is 'pVeg' which is a constitutive promoter. Finally, the RFP system is working very well because when leak occurs, cells glow very strongly.
 
 
 
{| border="1" class="wikitable" style="text-align: center;"
 
|+T7 autoloop in E.coli at 37°C
 
|-
 
|[[File:T7 autoloop ctrl-25.jpg|450px|thumb|center|E.coli T7 autoloop without terminator at 37°C  (trans image)]]
 
|[[File:T7 autoloop ctrl-26.jpg|450px|thumb|center|E.coli T7 autoloop without terminator at 37°C  (gfp image)]]
 
|-
 
|[[File:T7_loop_1.jpg|450px|thumb|center|E.coli T7 autoloop with terminator  at 37°C  (trans image)]]
 
|[[File:T7 loop 1 gfp.jpg|450px|thumb|center|E.coli T7 autoloop with terminator at 37°C  (gfp image)]]
 
|-
 
|[[File:T7 autoloop ctrl-10.jpg|450px|thumb|center|E.coli negative control  at 37°C  (trans image)]]
 
|[[File:T7 autoloop ctrl-11.jpg|450px|thumb|center|E.coli negative control at 37°C  (gfp image)]]
 
|}
 
 
These pictures show that the T7 GFP autoloop system is efficient since some cells are glowing with GFP fluorescence. Thus, we can conclude that the T7 polymerase is activated.
 
Finally, we notice there is no difference between the GFP autoloop with and without terminator before the T7 promoter, we can interpret this due to this terminator is a B.subtilis terminator, moreover, we know that this E.coli strain has 4 terminators.
 
 
===User Reviews===
 
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|-
 
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<I>Username</I>
 
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Enter the review inofrmation here.
 
|};
 
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Revision as of 03:21, 22 September 2011