Part:BBa_K2324006
pRha_FimH_225sfGFP
The literature has shown that the terminal pili protein FimH (Le Trong et al 2010) can be modified by inserting heterologous sequences at position 225 and 258 (Pallesen et al 1995, Shembri et al 1999). However these two amino acids are in the pilin binding domain which may present difficulties when attempting to introduce large modifications. Harvard iGEM 2015 also introduced modifications at position 225 of the mature FimH protein.
This part produces a FimH adhesin protein fused with sfGFP at its 225th amino acid residue, after cleavage of the signal peptide. The coding sequence is under the control of a rhamnose-inducible promoter, with a B0034 RBS and a B0015 terminator. The part, when induced, should produce a fluorescent FimH protein that should initiate pilus biosynthesis when co-transformed with a plasmid containing the fim operon. Unfortunately despite best efforts (multiple growth temperatures and multiple rhamnose inducing concentrations) no fluorescence has been detected in any of the E. coli strains.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 451
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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