Team:Heidelberg/Templates/Indigoidine week13 overview

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Latest revision as of 15:18, 21 October 2013

Indigoidine Production – Testing PPTases

We wanted to test various PPTases on their functionality in activating the indigoidine synthetases indC and bpsA. The plasmids pRB3-10 represent all possible combinations of either indC or bpsA with the PPTases sfp, svp, svp (pMM65) or entD. We transformed E. coli TOP10 with all the plasmids.

All PPTases are capable of attaching the 4'-Phosphopanthetheinyl residue to the apo-form of the indigoidine synthetases as was observed by the blue phenotype of transformed E. coli TOP10 cells. The main focus was now on exchanging the T-Domain of the indigoidine synthetase indC to further proof the concept of modularity in the field of NRPS domains. We can easily determine which engineered versions of indC remain functionality by screening for blue colonies.

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 ===Indigoidine Production – Testing PPTases===
 <p>
-We wanted to test various PPTases on their functionality in activating the indigoidine synthetases indC and bpsA. The plasmids [[Plasmids#pRB3|pRB3-10]] represent all possible combinations of either indC or bpsA with the PPTases [[Gene#sfp|sfp]], [[Gene#svp|svp]], [[Gene#svp|svp]] ([[Plasmids#pMM65|pMM65]]) or [[Gene#entD|entD]]. We transformed [[Strain#TOP10|''E. coli'' TOP10]] with all the plasmids.
+We wanted to test various PPTases on their functionality in activating the indigoidine synthetases indC and bpsA. The plasmids pRB3-10 represent all possible combinations of either indC or bpsA with the PPTases sfp, svp, svp (pMM65) or entD. We transformed ''E. coli'' TOP10 with all the plasmids.
 </p>
 <p>
-All [[IndPD#PPTase|PPTases]] are capable of attaching the 4'-Phosphopanthetheinyl residue to the apo-form of the indigoidine synthetases as was observed by the blue phenotype of transformed ''E. coli'' TOP10 cells. The main focus was now on exchanging the [[IndPD#T-Domain|T-Domain]] of the indigoidine synthetase [[Gene#indC|indC]] to further proof the concept of modularity in the field of NRPS domains. We can easily determine which engineered versions of indC remain functionality by screening for blue colonies.
+All PPTases are capable of attaching the 4'-Phosphopanthetheinyl residue to the apo-form of the indigoidine synthetases as was observed by the blue phenotype of transformed ''E. coli'' TOP10 cells. The main focus was now on exchanging the T-Domain of the indigoidine synthetase indC to further proof the concept of modularity in the field of NRPS domains. We can easily determine which engineered versions of indC remain functionality by screening for blue colonies.
 </p>