Difference between revisions of "Part:BBa K1741011"
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The sfGFP fluorescence [RFU] was measured using Tecan fluorometer.
 
The sfGFP fluorescence [RFU] was measured using Tecan fluorometer.
  
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As you can see, there is high expression of T7 promoter however there is no inducer (lactose) present in the medium (so it has a tendency to leak). Our promoters don't show any significant expression. Please note that this comparison is simplified, because the transcription from T7 promtoer is performed by strong viral polymerase while the transcription from our other promoters is performed by slower cellular RNA polymerase.
+
As you can see, there is high expression of T7 promoter however there is no inducer (lactose) present in the medium (so it has a tendency to leak). Our promoters don't show any significant expression. Please note that this comparison is simplified, because the transcription from T7 promoter is performed by strong viral polymerase while the transcription from our other promoters is performed by slower cellular RNA polymerase.
  
  

Revision as of 06:52, 27 September 2015

sfGFP under T7 promoter

This part imitates the most popular IPTG/lactose dependent T7 driven expression systems. To express sfGFP or another sequence with which one can substitute sfGFP ORF using Gibson or CPEC assembly, host strain of choice expressing T7 RNA polymerase is necessary, not only DE3.

Results

We compared our promoters to T7 promoter on minimal M9 medium with added glucose.

Teamuampoznandifferentpromotersm9minimal.png

The sfGFP fluorescence [RFU] was measured using Tecan fluorometer.

As you can see, there is high expression of T7 promoter however there is no inducer (lactose) present in the medium (so it has a tendency to leak). Our promoters don't show any significant expression. Please note that this comparison is simplified, because the transcription from T7 promoter is performed by strong viral polymerase while the transcription from our other promoters is performed by slower cellular RNA polymerase.



Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 48
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 48
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 48
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 131