Part:BBa_K2324015
Arabinose inducible fim operon
This part contains the fim operon under the control of an inducible arabinose promoter. The operon contains six Fim proteins, including FimA, FimI, FimC, FimD, FimF and FimG. When co-transformed with FimH constructs from our project, which have rhamnose-inducible promoters, modified type I pili may be produced. The production of the operon must be initiated by inducing the culture overnight at 0.6 OD at 2% arabinose.
Previous iGEM team, Harvard 2015 has synthesised a very similar part, however we used the modular cloning strategy, which has produced scar-sites at various sequences and it has also been codon optimised by the IDT E. coli codon optimisation software.
Results
Figure 1 Top10 with the fim operon under the control of promoter P_Ara showed no visible signs of pili expression with insertion of the operon alone.
Figure 2 We transformed a plasmid containing the fim operon under the control of the P_Ara promoter into ΔFimB. It exhibits flagella on their cell surface, but the electron micrograph also shows a suggestion of pili.
Results
This suggests that the P_Ara construct was successful in synthesising pili, without the co-transformation with a FimH construct.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 125
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 65
Illegal BamHI site found at 2743 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1026
Illegal AgeI site found at 1057 - 1000COMPATIBLE WITH RFC[1000]
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