Part:BBa_K2871001
EspD Translocon protein. Membrane protein for T3SS docking.
A Translocon is a complex of proteins that assemble a pore-like structure in membranes. EspD is part of a group of proteins used for assembly of a translocon that works like a docking site for the bacterial type-3-secretion system (T3SS). This docking site is also the site of protein injection by the T3SS.
Usage and Biology
Gene expression and Protein purification
EspD was PCR from SIEC E. coli strain and insert into IPTG inducible BBa_K731500 vector under pTac promoter and BBa_B0034 ribosome binding site by USER cloning. The expression vector map is shown in Figure 1. The expression vector is transformed into E. coli and the expression is induced with 1mM IPTG in LB broth. The cell was lysed by sonication and centrifugation. Western blot experiment shows that His-tagged EspD protein is only present in the pellet part of the lysed cell as shown in Figure 2. However, the amount of EspD protein is very low, which prohibit purification.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 422
Illegal XhoI site found at 1045 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 739
- 1000COMPATIBLE WITH RFC[1000]
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