Composite
FimH-Dterm

Part:BBa_K2052014:Design

Designed by: Burak Kızıl, Seniz Yuksel   Group: iGEM16_METU_HS_Ankara   (2016-09-24)


FimH site directed mutated with RPMrel


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1173
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1113
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Our protein coding region (FimH) are digested from our whole construct with the enzymes EcoRI and SpeII to clone into pSB1C3. Since it can be used for targetting teraphy, we thought that pBAD promoter may be useful for further studies of one who wanted to induce the protein expression after obtaining proper binding of FimH and can carry project to next level such as treatment or repression.

Source

Some substrains of E.Coli have a structure called Type 1 pili which is expressed from the Fim gene system. At the end of the pili structure there is a protein called “FimH” which is the structure that allows them to bind to the mannose sugar that is found on the surfaces of eukaryotic cells. (Sauer et al., 2016)

References

Krogfelt KA., Bergmans H., Klemm P. (1990, June) “Direct evidence that the FimH protein is the mannose-specific adhesin of Escherichia coli type 1 fimbriae”

Sauer MM, Jakob RP, Eras J, Baday S, Eriş D, Navarra G, Bernèche S, Ernst B, Maier T, Glockshuber R. (2016 March 7). “Catch-bond mechanism of the bacterial adhesin FimH”

Kelly, K. A., Jones, D. A., (2003). “Isolation of a Colon Tumor Specific Binding Peptide Using Phage Display Selection”

Figure 1. iGEM 2015 Harvard BioDesign . Retrieved from https://parts.igem.org/Part:BBa_K1850010

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1550331/


Induction protocol http://www.embl.de/pepcore/pepcore_services/protein_expression/ecoli/