Part:BBa_K3786006
FtsZ regulated by pLac
Usage and Biology
The ftsZ (filamenting temperature-sensitive mutant Z) gene is encoding an essential cell division protein: FtsZ. FtsZ is a tubulin-like protein that has a crucial role during bacterial division as this is the first cell-division protein to arrive on the division site (the septum of dividing cells). Moreover, its GTPase activity enables the contractile movement to promote cell division. FtsZ expression regulates the formation of a contractile ring structure also called ftsZ ring or Z ring, it consists of the assembly of protofilaments at the future cell division site.
We noticed that min mutant strains struggle to survive and therefore would not produce an optimum amount of minicells. Whereas by observing their limited lifespan, we could see that by not having a tight regulation of their expression we wouldn’t have clear knowledge of their age and therefore could not know how long they would still produce our proteins.
This led to the design of a tightly regulated minicell production loop. Our hypothesis was that by having a plasmid switching the bacteria on to produce minicells under specific conditions would increase minicells yield and ensure they would be still metabolically active to express protein of interest after purification assays.
Characterization
By adding 0.5mM of IPTG in the media, it was possible to observe a difference in the amount of minicells, from TB43 strain, transformed with the plasmid of FtsZ expression under IPTG promoter regulation. See Paris_Bettencourt 2021 wiki [1].
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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