Team:LMU-Munich/Inverter

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{! -- hier bitte das Bild vom Schaltkreis hin
{! -- hier bitte das Bild vom Schaltkreis hin
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The Inverter is composed of the promoter one wants to invert, in this proof of principle case pBAD, which is induced by arabinose, which regulates the small RNA RyhB. RyhB itself translationally inhibits uof (upstream of fur) by binding and masking the Shine Dalgarno sequence. If this is translationally fused to a reporter, in this case lacZalpha, this too is translationally repressed. The repression of the reporter is dependent on the concentration of RyhB, and therefore the Induction of the pBAD promoter. The Induction of the pBAD promoter is inverted to a negative output of the reporter.
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The [http://partsregistry.org/wiki/index.php?title=Part:BBa_K823040 Inverter] is composed of the promoter one wants to invert, in this proof of principle case pBAD, which is induced by arabinose, which regulates the small RNA RyhB. RyhB itself translationally inhibits uof (upstream of fur) by binding and masking the Shine Dalgarno sequence. If this is translationally fused to a reporter, in this case lacZalpha, this too is translationally repressed. The repression of the reporter is dependent on the concentration of RyhB, and therefore the Induction of the pBAD promoter. The Induction of the pBAD promoter is inverted to a negative output of the reporter.
The ONPG assay below shows the function of this Inverter. The Signal of the lacZalpha is so low, as the pBAD promoter lacks the repressor binding sites, and is therefore quite leaky, which makes it less titratable. But as this peticuliar Inverter is just a proof of principle, it is not essential. The Inverter for the wanted Promoter and Output has to be constructed by FusionsPCR.
The ONPG assay below shows the function of this Inverter. The Signal of the lacZalpha is so low, as the pBAD promoter lacks the repressor binding sites, and is therefore quite leaky, which makes it less titratable. But as this peticuliar Inverter is just a proof of principle, it is not essential. The Inverter for the wanted Promoter and Output has to be constructed by FusionsPCR.

Revision as of 11:57, 25 September 2012

iGEM Ludwig-Maximilians-Universität München Beadzillus

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The LMU-Munich team is exuberantly happy about the great success at the World Championship Jamboree in Boston. Our project Beadzillus finished 4th and won the prize for the "Best Wiki" (with Slovenia) and "Best New Application Project".

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