Difference between revisions of "Part:BBa K1741006"

 
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<partinfo>BBa_K1741006 short</partinfo>
 
<partinfo>BBa_K1741006 short</partinfo>
  
 
From rhaBAD1 promoter we removed both CRP binding sites. It seems likely thatt RhaS binding site has been also affected, so however the resulting promoter is still induced by rhamnose, the induction is very weak.
 
From rhaBAD1 promoter we removed both CRP binding sites. It seems likely thatt RhaS binding site has been also affected, so however the resulting promoter is still induced by rhamnose, the induction is very weak.
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===Characteristic and Results===
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Legend:
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RhaWT - rhamnose wild type
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Rha1 - [[https://parts.igem.org/Part:BBa_K1741005 BBa_K1741005]]
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Rha2 - [[https://parts.igem.org/Part:BBa_K1741006 BBa_K1741006]]
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https://static.igem.org/mediawiki/parts/9/92/Teamuampoznanrhamnoselbgraph.png
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https://static.igem.org/mediawiki/parts/0/0d/Teamuampoznanrhamnosem9graph.png
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The sfGFP fluorescence [RFU] was measured using Tecan fluorometer.
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All three rhamnose promoters are induced by rhamnose and repressed by glucose.
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Turns out that single point mutation in EcoRI restriction site in Rha1 significantly increased the activity of the promoter as comapred to the wild type construct.
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Rha2 is a weak promoter, perfect for people who would like to manufacture small amount of the protein of interest.
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We also checked the tightness of our promoters.
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All of our promoters are induced only by their respective sugars with a small exception of xylose promoters being slightly induced by arabinose and arabinose promoters being slightly induced by xylose. All of our promoters are repressed by glucose.
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https://static.igem.org/mediawiki/parts/4/43/Teamuampoznanpromoterscomparisongrpah.png
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 05:47, 27 September 2015

sfGFP under short rhamnose promoter (very weak)

From rhaBAD1 promoter we removed both CRP binding sites. It seems likely thatt RhaS binding site has been also affected, so however the resulting promoter is still induced by rhamnose, the induction is very weak.

Characteristic and Results

Legend:

RhaWT - rhamnose wild type

Rha1 - [BBa_K1741005]

Rha2 - [BBa_K1741006]

Teamuampoznanrhamnoselbgraph.png Teamuampoznanrhamnosem9graph.png


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

All three rhamnose promoters are induced by rhamnose and repressed by glucose. Turns out that single point mutation in EcoRI restriction site in Rha1 significantly increased the activity of the promoter as comapred to the wild type construct. Rha2 is a weak promoter, perfect for people who would like to manufacture small amount of the protein of interest.

We also checked the tightness of our promoters.

All of our promoters are induced only by their respective sugars with a small exception of xylose promoters being slightly induced by arabinose and arabinose promoters being slightly induced by xylose. All of our promoters are repressed by glucose. Teamuampoznanpromoterscomparisongrpah.png



Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 148