Difference between revisions of "Part:BBa K2324001"
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<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 225 of the mature FimH protein.</p>
 
<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 225 of the mature FimH protein.</p>
 
<p>
 
<p>
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CDS which codes for a FimH protein fused with a sfGFP at residue 225 after cleavage of the signal peptide. This part tests expression and folding of FimH when modified by a large protein position 225, after signal peptide cleavage. 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_K2324011 under the T7 promoter, as well as https://parts.igem.org/Part:BBa_K2324006 under the pRha promoter. </p>
+
CDS which codes for a FimH protein fused with a sfGFP at residue 225 after cleavage of the signal peptide. This part tests expression and folding of FimH when modified by a large protein position 225, after signal peptide cleavage. This part is included in the composite part https://parts.igem.org/Part:BBa_K2324011 under the T7 promoter, as well as https://parts.igem.org/Part:BBa_K2324006 under the pRha promoter. </p>
  
 
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Revision as of 21:17, 1 November 2017


FimH+sfGFP at residue 225

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.

CDS which codes for a FimH protein fused with a sfGFP at residue 225 after cleavage of the signal peptide. This part tests expression and folding of FimH when modified by a large protein position 225, after signal peptide cleavage. This part is included in the composite part https://parts.igem.org/Part:BBa_K2324011 under the T7 promoter, as well as https://parts.igem.org/Part:BBa_K2324006 under the pRha promoter.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 185
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]