Difference between revisions of "Team:Exeter/Basic Part"

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         <h4 id="h3">Fim Operon</h4>
 
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           <p>The fim operon consists of six proteins with a native RBS, it was produced by a previous iGEM team from Harvard in 2015.</p>  
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The fim operon consists of six proteins and their native RBS sequences. For synthesis the operon was split into three separate parts. It was produced by a previous iGEM team from Harvard in 2015.</p>  
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Revision as of 14:10, 29 October 2017

FimH sfGFP

To easily monitor expression we chose to modify FimH by inserting the super folder green fluorescent protein (sfGFP) (Pedelacq et al 2005). The sfGFP was inserted in three different places in the FimH protein to act as a reporter on the level of expression of the protein and thus giving an indication of its efficiency. The three places of insertion were 22, which is the place where the signal peptide ends in the mannose binding domain, the 225 and 258, which are present in the domain that interacts with FimG.

Name Description Base Pairs
BBa_K2324002 FimH_22_sfGFP 1617
BBa_K2324001 FimH_225_sfGFP 1617
BBa_K2324003 FimH_258_sfGFP 1617

FimH Fusion Proteins

We initially chose three metal binding proteins in order to bind a variety of metal ions: Mouse Metallothionein (you need to find a reference for this) to bind Cd, Cu and Zn; Synechococcus Metallothionein (Blindauer et al 2001) to bind cadmium and zinc; and Synechococcus Plastocyanin (Inoue et al 1999) to bind copper. We also chose to insert a 6x histidine tag to act as both a reporter of expression but also to bind Ni. Unfortunately no construct was successfully built containing plastocyanin.

Name Description Base Pairs
BBa_K2324014 FimH_22_His 921
BBa_K2324004 FimH_22_SynMT 1071
BBa_K2324005 FimH_22_MouseMT 1086

Fim Operon

The fim operon consists of six proteins and their native RBS sequences. For synthesis the operon was split into three separate parts. It was produced by a previous iGEM team from Harvard in 2015.

Name Description Base Pairs
Part:BBa_K1850012 Fim Operon 5744