Part:BBa_K4722012
BrkA
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 313
Illegal NgoMIV site found at 742
Illegal NgoMIV site found at 1378 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1346
Usage and Biology
BrkA, short for Bordetella Serum Killer Resistance Protein A, represents an autotransporter protein originating from the pertussis bacterium[1]. Within the repertoire of virulence factors produced by pertussis, BrkA plays a prominent role. This protein functions as a safeguard against cleavage by the canonical complement pathway, thereby contributing significantly to resistance. In a notable development, a novel autotransporter, designated as BrkAu, has emerged as an anchor for a bacterial surface display system. This system, termed BrkAutoDisplay, draws inspiration from the structural framework of BrkA. A distinguishing feature of BrkA is its capacity to retain the cleaved passenger domain firmly affixed to the bacterial surface. This characteristic facilitates the stable association of exogenous proteins with the bacterial cell, thereby enabling the convenient recycling and reutilization of functionalized bacteria.
Design Consideration
The genetic construct was ligated into a pET28a plasmid vector and subsequently introduced into Escherichia coli strain BL21 (DE3). NicX was genetically connected with BrkA to enable its direct translation onto the surface of BL21. This innovation eliminated the need for protein purification steps, allowing for the direct utilization of E. coli as a host for enzymes in various applications.
References
- ↑ Sun, F., Pang, X., Xie, T., Zhai, Y., Wang, G., & Sun, F. (2015). BrkAutoDisplay: functional display of multiple exogenous proteins on the surface of Escherichia coli by using BrkA autotransporter. Microbial Cell Factories, 14, 1-12. https://doi.org/10.1186/s12934-015-0316-3
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