Part:BBa_K257005
TolR domain II N-term fusion
TolR is known to play a role in the Tol/Pal membrane integrity system TolRII is a short part of the TolRII coding sequenced and it destabilize the membrane integrity that allow us to see vesicles formation
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]
Sequence Update by UNSW (2016)
Like Paris (2009), we were also interested in using Outer Membrane Vesicles (OMVs) in our project. We wanted to express this part to stimulate OMV production in E. coli, but came across an issue - this part lacks a start codon. While this may be optimal for use with many expression plasmids, it makes 3A assembly it bit trickier as an ATG has to be incorporated in somehow.
A second issue was that there's only a single stop codon, whereas double stop codons can be favoured for more certainty in terminating translation.
We thus redesigned the sequence to introduce these two new features - an extra start, and an extra stop - and submitted it as part BBa_K2022002. We also created BBa_K2022003, which adds a his-tag just prior to the double stop, enabling easier antibody-based recognition
We then characterised this part, and have evidence to suggest this part works as advertised, i.e. that it helps induce vesicle formation
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