Difference between revisions of "Part:BBa K2324003"
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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 258 of the mature FimH protein.</p>
 
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 258 of the mature FimH protein.</p>
 
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<p>
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This part tests expression and folding of FimH. Fluorescence suggests both that the protein is sufficiently folded such that the sfGFP works. Other techniques can then be used to demonstrate successful pilus biosynthesis. This part is included in the composite part https://parts.igem.org/Part:BBa_K2324008 under the pRha promoter.</p>
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This part tests expression and folding of FimH when modified by a large protein at position 258. Unfortunately despite best efforts (multiple growth temperatures and multiple rhamnose inducing concentrations) no fluorescence has been detected in any of the <i>E. coli</i> strains. This part is included in the composite part https://parts.igem.org/Part:BBa_K2324008 under the pRha promoter.</p>
  
 
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Revision as of 21:17, 1 November 2017


FimH+sfGFP at residue 258

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 258 of the mature FimH protein.

This part tests expression and folding of FimH when modified by a large protein at position 258. Unfortunately despite best efforts (multiple growth temperatures and multiple rhamnose inducing concentrations) no fluorescence has been detected in any of the E. coli strains. This part is included in the composite part https://parts.igem.org/Part:BBa_K2324008 under the pRha promoter.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]