Part:BBa_K3128030
OmpX gene without signal peptide
OmpX is an outer membrane protein with the C- and N-termini in the intracellulair domain. (BBa_K1761000) iGEM Grenoble designed this biobrick in order to add a leucine zipper to the C-terminal end of OmpX muted. This basic biobrick is a part of the positive control system of the iGEM Grenoble-Alpes 2019 project (see [http://2019.igem.org/Team:Grenoble-Alpes] for more informations.
Gene Design
In the Eindhoven 2015 team project ([http://2015.igem.org/Team:TU_Eindhoven/Project/Design]), this protein was modified with a Y121F' mutation.
To be able to use OmpX as a scaffold, one needs to integrate the unnatural amino acid. To do so, some modifications must be made to this part. Within the original OmpX part, a codon was mutated into the amber stop codon TAG. This codon will be incorporated in the protruding loops of OmpX, loop 2 and loop 3 (see Figure 1).
Figure 1: A) The OmpX protein structure has been elucidated through NMR and X-ray crystallography, B) The square residues are important for the secondary structure of OmpX. To keep the structure intact, we introduce an amber stop codon in one of the protruding loops. Figure 5B is adapted from [1].
These loops were chosen because they are easily accessible and are not part of the beta-barrel of OmpX [3]. Mutations in these loops may not disturb the secondary structure of OmpX. With a specific tRNA it is possible to implement a unnatural amino acid at the place of the TAG codon.
This biobrick not contains the putative signal peptide. More informations in the design page of the part.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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