Difference between revisions of "Part:BBa K2324004"

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

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 879
Illegal AgeI site found at 306
1000
COMPATIBLE WITH RFC[1000]

@@ Line 2: / Line 2: @@
 __NOTOC__
 <partinfo>BBa_K2324004 short</partinfo>
+<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>
+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>
-This part codes for a FimH adhesin fused with a metallothionein binding domain from <i>Synechococcus elongatus </i> at amino acid residue 1. 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 a number of heavy metal ions.
+<h2>References </h2>
+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.
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