Not Released
Experience: Works
Not Used
Get This Part
Coding
SgsE

Part:BBa_K525303

Designed by: Timo Wolf   Group: iGEM11_Bielefeld-Germany   (2011-09-10)

S-Layer SgsE from Geobacillus stearothermophilus with T7/lac

Bielefeld-Germany2011-S-Layer-Geometrien.jpg

S-Layer SgsE from Geobacillus stearothermophilus

S-layers (crystalline bacterial surface layer) are crystal-like layers consisting of multiple protein monomers and can be found in various (archae-)bacteria. They constitute the outermost part of the cell wall. Especially their ability for self-assembly into distinct geometries is of scientific interest. At phase boundaries, in solutions and on a variety of surfaces they form different lattice structures. The geometry and arrangement is determined by the C-terminal self assembly-domain, which is specific for each S-layer protein. The most common lattice geometries are oblique, square and hexagonal. By modifying the characteristics of the S-layer through combination with functional groups and protein domains as well as their defined position and orientation to eachother (determined by the S-layer geometry) it is possible to realize various practical applications (Sleytr et al., 2007).


Usage and Biology

S-layer proteins can be used as scaffold for nanobiotechnological applications and devices by e.g. fusing the S-layer's self-assembly domain to other functional protein domains. It is possible to coat surfaces and liposomes with S-layers. A big advantage of S-layers: after expressing in E. coli and purification, the nanobiotechnological system is cell-free. This enhances the biological security of a device.

This S-layer gene is downsteam of a T7 / lac fusion promoter and in the Freiburg BioBrick Assembly standard so functional protein domains can easily be fused to its N-terminus and expressed and purified afterwards. SgsE was characterized with BBa_K525305.


Important parameters

Experiment Characteristic Result
Expression (E. coli) Localisation Inclusion body
Compatibility E. coli KRX and BL21(DE3)
Induction of expression expression of T7 polymerase + IPTG or lactose
Purification Molecular weight 83.2 kDa
Theoretical pI 5.75
Immobilization behaviour Immobilization time 4 h


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 167
    Illegal BglII site found at 1022
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 76
    Illegal AgeI site found at 2401
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1657


[edit]
Categories
//chassis/prokaryote/ecoli
//proteindomain/internal
//rnap/bacteriophage/t7
Parameters
None