Difference between revisions of "Part:BBa K606040:Experience"

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The Hyperspank promoter was designed to be compatible for B. subtilis. It is shorter thant the classic one, but is also recognised by E. coli polymerases.
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The Hyperspank promoter was designed to be compatible for B. subtilis. It is shorter than the classic one, but is also recognised by E. coli polymerases.
  
We cloned it first behind the amber tRNA that would induce a GFP amber on an other plasmid. Then to characterise it we cloned it behind an RFP, into TURBO cells that are LacIq to allow the promoter to be the less leacky possible.
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We characterised it with an RFP expression system (PHs-RBS-RFP), into TURBO cells that express LacI to allow the promoter to be the less leacky possible. This system is inducible by IPTG.
  
 
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Revision as of 18:26, 23 September 2011


The Hyperspank promoter was designed to be compatible for B. subtilis. It is shorter than the classic one, but is also recognised by E. coli polymerases.

We characterised it with an RFP expression system (PHs-RBS-RFP), into TURBO cells that express LacI to allow the promoter to be the less leacky possible. This system is inducible by IPTG.



Without IPTG


E.coli containing Hyperspank+RBS+RFP - IPTG at 37°C
-IPTG E.coli at 37°C (rfp image)
-IPTG E.coli at 37°C (trans image)
-IPTG E.coli at 37°C (rfp image)
-IPTG E.coli at 37°C (trans image)


With IPTG :

E.coli containing Hyperspank+RBS+RFP + IPTG at 37°C
+IPTG E.coli at 37°C (rfp image)
+IPTG E.coli at 37°C (trans image)
+IPTG E.coli at 37°C (rfp image)
+IPTG E.coli at 37°C (trans image)

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