Difference between revisions of "Part:BBa K2324010"
 
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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 1 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 1 of the mature FimH protein.</p>
 
<p>
 
<p>
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This part produces a FimH adhesin protein fused with a metallothionein from <i>Mus musculus</i> at the first amino acid position. The coding sequence is under the control of a rhamnose-inducible promoter, with a B0034 RBS and a B0015 terminator. The part, when induced, produces a metal binding FimH protein that should initiates pilus biosynthesis when co-transformed with a plasmid containing the <i>fim operon</i></p> .
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This part produces a FimH adhesin protein fused with a metallothionein from <i>Mus musculus</i> (Huang et al 1981) at the first amino acid position, after signal peptide cleavage. The coding sequence is under the control of a rhamnose-inducible promoter (BBa_K902065), with the BBa_B0034 RBS and the BBa_B0015 terminator. The part, when induced, produces a metal binding FimH protein that should initiates pilus biosynthesis when co-transformed with a plasmid containing the <i>fim operon</i> Unfortunately, despite best efforts we were unable to characterise this part.</p>
  
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<h2>References </h2>
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Huang, I-Y., Kimura, M., Hata A., Tsunoo H., Yoshida A. (1981) Complete Amino Acid Sequence of Mouse Liver Metallothionein-II. Biochemistry 89, 1839-1845.
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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.
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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.
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Schembri, M. A., Kjaergaard, K., and KLEMM, P. (1999) Bioaccumulation of heavy metals by fimbrial designer adhesins. FEMS Microbiology Letters 170, 363–371.
  
 
<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>

Latest revision as of 22:11, 1 November 2017


pRha_FimH_1MouseMT

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 produces a FimH adhesin protein fused with a metallothionein from Mus musculus (Huang et al 1981) at the first amino acid position, after signal peptide cleavage. The coding sequence is under the control of a rhamnose-inducible promoter (BBa_K902065), with the BBa_B0034 RBS and the BBa_B0015 terminator. The part, when induced, produces a metal binding FimH protein that should initiates pilus biosynthesis when co-transformed with a plasmid containing the fim operon Unfortunately, despite best efforts we were unable to characterise this part.

References

Huang, I-Y., Kimura, M., Hata A., Tsunoo H., Yoshida A. (1981) Complete Amino Acid Sequence of Mouse Liver Metallothionein-II. Biochemistry 89, 1839-1845.

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
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]