Part:BBa_K3788027
NOTOC pelB_sfGFP (without stop) _ aida-I (D33:N)
BBa_K3788026 is coding for pelB, sfGFP and AidaA. pelB is a leader sequence, it allows the protein to join the periplasm via the SEC system (unfolded proteins). Aida-I, on the other hand, is an autotransporter from the T5SS; it allow the addressing to the extracellular medium, the mutation of the 846 – 847 amino acid permit to no have protein cleavage, and attach this one to the extracellular membrane(1). In this contruction sfGFP permit to understand the mechanism, this protein can at the end be replaced by an other protein. sfGFP is supposed to be address to the preriplasm thanks to pelB leader and the T5SS system will address sfGFP to the extracellular membrane. Sequence was optimised for E. coli K12.
Figure coming from The Autodisplay Story, from Discovery to Biotechnical and Biomedical Applications
A. representation of the mRNA coding for an autotransporter system.
B. representation of the events of translation, folding in the periplasm and export of the passenger sequence to the extracellular medium.
This figure explains how is constituted a gene coding for an autotransporter such as aidaI. We replaced the signal peptide (SP) by PelB leader sequence and the passenger sequence by sfGFP sequence so that this protein should be transported to the periplasm and then pass to the extracellular medium by the autotransporter AidaI.
(1) : https://www.sciencedirect.com/science/article/pii/S0021925818665512
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 54
Illegal AgeI site found at 1864 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 79
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