Difference between revisions of "Part:BBa K2324004"
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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 1 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 1 of the mature FimH protein.</p> | ||
<p> | <p> | ||
| − | This part codes for a FimH adhesin fused with a metallothionein protein from <i>Synechococcus elongatus </i> ( | + | This part codes for a FimH adhesin fused with a metallothionein protein from <i>Synechococcus elongatus </i> (Blindauer et al 2001) at amino acid residue 1, after signal peptide cleavage. It is designed to be placed on the end of a type 1 pilus structure in <i> E. coli </i> with a view to binding cadmium and zinc metal ions. This part is included in the composite part https://parts.igem.org/Part:BBa_K2324009 under the pRha promoter. </p> |
<h2>References </h2> | <h2>References </h2> | ||
Latest revision as of 22:05, 1 November 2017
FimH+SynMT
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 1 of the mature FimH protein.
This part codes for a FimH adhesin fused with a metallothionein protein from Synechococcus elongatus (Blindauer et al 2001) at amino acid residue 1, after signal peptide cleavage. It is designed to be placed on the end of a type 1 pilus structure in E. coli with a view to binding cadmium and zinc metal ions. This part is included in the composite part https://parts.igem.org/Part:BBa_K2324009 under the pRha promoter.
References
Blindauer, C. A., Harrison, M. D., Parkinson, J. A., Robinson, A. K., Cavet, J. S., Robinson, N. J., and Sadler, P. J. (2001) A metallothionein containing a zinc finger within a four-metal cluster protects a bacterium from zinc toxicity. Proc. Natl. Acad. Sci. U.S.A. 98, 9593–9598.
Le Trong, I., Aprikian, P., Kidd, B. A., Forero-Shelton, M., Tchesnokova, V., Rajagopal, P., Rodriguez, V., Interlandi, G., Klevit, R., Vogel, V., Stenkamp, R. E., Sokurenko, E. V., and Thomas, W. E. (2010) Structural Basis for Mechanical Force Regulation of the Adhesin FimH via Finger Trap-like Sheet Twisting. Cell 141, 645–655.
Pallesen, L., Poulsen, L. K., Christiansen, G., and Klemm, P. (1995) Chimeric Fimh Adhesin of Type-1 Fimbriae - a Bacterial Surface Display System for Heterologous Sequences. Microbiology 141, 2839–2848.
Schembri, M. A., Kjaergaard, K., and KLEMM, P. (1999) Bioaccumulation of heavy metals by fimbrial designer adhesins. FEMS Microbiology Letters 170, 363–371.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 879
Illegal AgeI site found at 306 - 1000COMPATIBLE WITH RFC[1000]