Coding
SbpA | Luc

Part:BBa_K525411

Designed by: Anna Drong   Group: iGEM11_Bielefeld-Germany   (2011-09-13)

Fusion Protein of S-Layer SbpA and Firefly-Luciferase

Bielefeld-Germany2011-S-Layer-Geometrien.jpg

Fusion protein of S-layer SbpA and firefly luciferase.

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 fusion protein is used to characterize purification methods and immobilization behaviour of enzymes fused to an S-layer.


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 170.6 kDa
Theoretical pI 5.14
Reporter Luminescence
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 104
    Illegal BglII site found at 221
    Illegal XhoI site found at 1996
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 76
    Illegal AgeI site found at 4846
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 493
    Illegal BsaI.rc site found at 622


[edit]
Categories
//chassis/prokaryote/ecoli
//function/reporter/light
//proteindomain/internal
Parameters
biologyS-Layer
chassisE. coli
controlPT7
originLysinibacillus sphaericus