Fusion Protein of S-Layer SbpA and Firefly-Luciferase
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.
|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|
|Immobilization behaviour||Immobilization time||4 h|
Sequence and Features
- 10COMPATIBLE WITH RFC
- 12COMPATIBLE WITH RFC
- 21Illegal BglII site found at 104
Illegal BglII site found at 221
Illegal XhoI site found at 1996
- 23COMPATIBLE WITH RFC
- 25Illegal NgoMIV site found at 76
Illegal AgeI site found at 4846
- 1000Illegal BsaI.rc site found at 493
Illegal BsaI.rc site found at 622