Difference between revisions of "Part:BBa K902066"
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[[Image:Calgary_RhaGFPFinal.png|thumb|500px|center|GFP expression for cells expressing Prha-RBS-GFP that were incubated under different conditions. We can see a large increase with 0.2% rhamnose, 0.5% rhamnose whereas there is no GFP expression in the cells incubated with glucose as compaed to our control.]]
 
[[Image:Calgary_RhaGFPFinal.png|thumb|500px|center|GFP expression for cells expressing Prha-RBS-GFP that were incubated under different conditions. We can see a large increase with 0.2% rhamnose, 0.5% rhamnose whereas there is no GFP expression in the cells incubated with glucose as compaed to our control.]]
  
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This results were very promising.  This graph indicates that not only can we induce induce expression rhamnose, but the promoter is quite tightly regulated with little leaky expression over a basal level.  This makes it an excellent candidate for a killswitch construct, as it would allow for veyr little leaky expression of any kill genes when not wanted and an easy way of inducing the kill gene expression.  We later tested this promoter out with an <I>S7 micrococcal nuclease</i>.  This data can be found [[https://parts.igem.org/wiki/index.php?title=Part:BBa_K902084 here]]  
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This results were very promising.  This graph indicates that not only can we induce induce expression rhamnose, but the promoter is quite tightly regulated with little leaky expression over a basal level.  This makes it an excellent candidate for a killswitch construct, as it would allow for very little leaky expression of any kill genes when not wanted and an easy way of inducing the kill gene expression.  We later tested this promoter out with an <I>S7 micrococcal nuclease</i>.  This data can be found [https://parts.igem.org/wiki/index.php?title=Part:BBa_K902084 here]
  
 
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Revision as of 06:49, 28 October 2012

pRha-B0034-K082003

Encodes for the pRha, RBS site and a GFP with an LVA tag. This circuit is used to characterize expression of the rhamnose promoter with GFP containing a degradation tag. Please view our [http://2012.igem.org/Team:Calgary/Project/HumanPractices/Killswitch/Regulation#rhamnose Wiki] for more details about this component of our project.

GFP expression for cells expressing Prha-RBS-GFP that were incubated under different conditions. We can see a large increase with 0.2% rhamnose, 0.5% rhamnose whereas there is no GFP expression in the cells incubated with glucose as compaed to our control.

This results were very promising. This graph indicates that not only can we induce induce expression rhamnose, but the promoter is quite tightly regulated with little leaky expression over a basal level. This makes it an excellent candidate for a killswitch construct, as it would allow for very little leaky expression of any kill genes when not wanted and an easy way of inducing the kill gene expression. We later tested this promoter out with an S7 micrococcal nuclease. This data can be found here

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 BsaI.rc site found at 910