Part:BBa_K5109023
Deha2 surface display system
This part is a modification of the expression cassette BBa K5109001 which has been improved by incorporating a sequence encoding the anchoring motif BBa K5109004, creating a display system. This expression cassette allows the display of an otherwise intracellular protein (Dehalogenase type II) on the cell surface by fusing it to a carrier.
Carrier of choice was Lpp - OmpT, with the addition of a linker, described as part: BBa K5109004. The choice of this linker compared to others was made based on previous bibliographic research to find out which pattern would offer the best surface expression. We introduced different restriction sites both upstream and downstream the carrier and the dehalogenase sequences, enabling the cassette's use in various projects. Specifically, by cutting with NdeI, the Lpp-OmpT sequence can be removed, in order to either convert the cassette to an intracellular expression one, or to allow the replacement of the carrier. In the latter case, it could be useful to compare the effectiveness of Lpp-OmpT as a carrier against other anchoring motifs. Additionally, the passenger protein can be exchanged by digesting with BsaI and BamHI.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 639
Illegal BamHI site found at 1372 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 667
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