Difference between revisions of "Part:BBa K4375012"
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<partinfo>BBa_K4375012 short</partinfo>
 
<partinfo>BBa_K4375012 short</partinfo>
  
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This protein contains the N-terminal domain of intimin, which is a bacterial adhesion factor, D0 an ig-like protein, and an E epitope tag. Intimin beta-barrel transmembrane domain anchors the construct into the outer membrane of the bacteria.  
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This protein contains the N-terminal domain of intimin, which is a bacterial adhesion factor, D0 an Ig-like protein, and an E epitope tag. Intimin is a β-barrel transmembrane domain that anchors the construct into the outer membrane of the bacteria.
  
  
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==Usage and Biology==
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<html>
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</p>
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</html>
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__TOC__
  
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Intimin families are large, secreted polypeptides that contain three functional regions: i) a N-terminal signal peptide, that drives their Sec-dependent translocation across the inner membrane; ii) a β-domain, that is anchored into the outer membrane and comprises a 12-stranded β-barrel with a peptide linker running through the lumen of the β-barrel; and iii) a passenger region, that is secreted to the extracellular milieu. Although their mechanism of secretion remains uncertain, Intimin proteins are translocated into the periplasm and then use the β-barrel assembly machine (BAM) complex for insertion into the outer membrane and translocation of the passenger region to the cell surface. The passenger region is found in the C-terminus. The β-barrel of Intimin proteins is followed by a peptide linker that runs through the lumen connecting its C-terminus to the passenger region.
 
  
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==Usage and Biology==
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Intimin, a bacterial adhesin, is an outer membrane protein used in bacterial display systems, and it consists of three regions: signal peptide, passenger domain, and a conserved, C-terminal translocator domain. This has a characteristic 12-stranded β-barrel structure, while the passenger domain acts as an autochaperon, which is absent in this part, except its initial region, D0. The structure of the barrel allows it to be an anchor in the membrane and proteins can be fused to it. Intimin have been used in several E.Coli based display systems.
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In this part, E-tag is fused to this protein, which is a 13 aa long peptide sequence, commonly used in biological applications to label proteins. Therefore, this construct could be labeled by immunostaining.
  
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==Sequence and Features==
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K4375012 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K4375012 SequenceAndFeatures</partinfo>
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==References==
 
==References==
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Leo, J. C.; Oberhettinger, P.; Schütz, M.; Linke, D. The Inverse Autotransporter Family: Intimin, Invasin and Related Proteins. International Journal of Medical Microbiology '''2015''', 305 (2), 276–282. https://doi.org/10.1016/j.ijmm.2014.12.011.
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Wentzel, A.; Christmann, A.; Adams, T.; Kolmar, H. Display of Passenger Proteins on the Surface of Escherichia Coli K-12 by the Enterohemorrhagic E. Coli  Intimin EaeA. Journal of Bacteriology '''2001''', 183 (24), 7273–7284. https://doi.org/10.1128/jb.183.24.7273-7284.2001.
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Rutherford, N., Mourez, M. Surface display of proteins by Gram-negative bacterial autotransporters. Microb Cell Fact 5, 22 '''2006'''. https://doi.org/10.1186/1475-2859-5-22
  
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https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0075126#
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Salema, V.; Marín, E.; Martínez-Arteaga, R.; Ruano-Gallego, D.; Fraile, S.; Margolles, Y.; Teira, X.; Gutierrez, C.; Bodelón, G.; Fernández, L. Á. Selection of Single Domain Antibodies from Immune Libraries Displayed on the Surface of E. Coli Cells with Two β-Domains of Opposite Topologies. PLoS ONE '''2013''', 8 (9), e75126. https://doi.org/10.1371/journal.pone.0075126.
  
  

Latest revision as of 19:45, 10 October 2022


Intimin domain for surface display (with anti-E epitope tag)

This protein contains the N-terminal domain of intimin, which is a bacterial adhesion factor, D0 an Ig-like protein, and an E epitope tag. Intimin is a β-barrel transmembrane domain that anchors the construct into the outer membrane of the bacteria.



Usage and Biology

Intimin, a bacterial adhesin, is an outer membrane protein used in bacterial display systems, and it consists of three regions: signal peptide, passenger domain, and a conserved, C-terminal translocator domain. This has a characteristic 12-stranded β-barrel structure, while the passenger domain acts as an autochaperon, which is absent in this part, except its initial region, D0. The structure of the barrel allows it to be an anchor in the membrane and proteins can be fused to it. Intimin have been used in several E.Coli based display systems. In this part, E-tag is fused to this protein, which is a 13 aa long peptide sequence, commonly used in biological applications to label proteins. Therefore, this construct could be labeled by immunostaining.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1983
    Illegal BamHI site found at 3999
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 2004
    Illegal NgoMIV site found at 4020
  • 1000
    COMPATIBLE WITH RFC[1000]


References

Leo, J. C.; Oberhettinger, P.; Schütz, M.; Linke, D. The Inverse Autotransporter Family: Intimin, Invasin and Related Proteins. International Journal of Medical Microbiology 2015, 305 (2), 276–282. https://doi.org/10.1016/j.ijmm.2014.12.011.

Wentzel, A.; Christmann, A.; Adams, T.; Kolmar, H. Display of Passenger Proteins on the Surface of Escherichia Coli K-12 by the Enterohemorrhagic E. Coli  Intimin EaeA. Journal of Bacteriology 2001, 183 (24), 7273–7284. https://doi.org/10.1128/jb.183.24.7273-7284.2001.

Rutherford, N., Mourez, M. Surface display of proteins by Gram-negative bacterial autotransporters. Microb Cell Fact 5, 22 2006. https://doi.org/10.1186/1475-2859-5-22

Salema, V.; Marín, E.; Martínez-Arteaga, R.; Ruano-Gallego, D.; Fraile, S.; Margolles, Y.; Teira, X.; Gutierrez, C.; Bodelón, G.; Fernández, L. Á. Selection of Single Domain Antibodies from Immune Libraries Displayed on the Surface of E. Coli Cells with Two β-Domains of Opposite Topologies. PLoS ONE 2013, 8 (9), e75126. https://doi.org/10.1371/journal.pone.0075126.