Difference between revisions of "Team:Exeter/Lab Introduction"
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<figcaption>Fig.1 Plasmids</figcaption> | <figcaption>Fig.1 Plasmids</figcaption> | ||
| + | <p>The FimH protein was modified to carry either sfGFP (reporter), 6xHistidine tag (reporter and metal binding) or two different metallothioneins (metal binding) (Fig.2a). See <a href="https://2017.igem.org/Team:Exeter/Basic_Part">Basic Parts</a> page for information. The Fim operon consisting of six proteins with native RBS (Fig.2b). Four different promoters were chosen to allow investigation of expression in a range of <i>E. coli</i> chassis. See Composite Parts page for information. Together this design gives a collection of parts for modified pili production. </p> | ||
<h5 id="h2">Hydrocyclone </h5> | <h5 id="h2">Hydrocyclone </h5> | ||
Revision as of 16:21, 27 October 2017
Pili+
Modified pili expression
We wanted to produce modified pili to primarily bind a variety of metal ions but also with reporters to allow easy verification of protein expression. A two plasmid system was designed where the modified FimH would be expressed from one plasmid and the remaining proteins in the fim operon expressed from the second plasmid. The two plasmids carried compatible origins of replication (pUC and p15A) and different antibiotic resistance genes (AmpR and CmR) to allow for co-transformation in E. coli (Fig.1)
The FimH protein was modified to carry either sfGFP (reporter), 6xHistidine tag (reporter and metal binding) or two different metallothioneins (metal binding) (Fig.2a). See Basic Parts page for information. The Fim operon consisting of six proteins with native RBS (Fig.2b). Four different promoters were chosen to allow investigation of expression in a range of E. coli chassis. See Composite Parts page for information. Together this design gives a collection of parts for modified pili production.
