Part:BBa_K4375012

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 peptide commonly used in biological applications to label proteins. Antibodies targeting this tag can be used in several techniques like ELISA or Western-blotting.

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.