Part:BBa_K3786000

Filamenting temperature-sensitive mutant Z

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.

Characterization

Studies that use FtsZ to induce cellular division for cell biology are very important for projects. By using this protein as a standardized coding sequence, we could evaluate minicell production through differentes periods of time and under regulation of different promoters. Depending on the simplified use of these parts, standardization allows the use of a single FtsZ gene in a coding sequence to a compound of synthetic biology in the form of biobricks. To produce minicells, Paris_Bettencourt 2021[1]team used this gene as one of it's approaches to induce minicell formation.

In the gel image, we can see successful amplification of FtsZ protein, expected to have a lenght of 1.2kb. The ladder used was 1kb Promega ladder. The gel was 1% agarose.

Figure 1. Gel Imaging with Syber Safe

Additionally, for understanding FtsZ role in production of minicells, we conducted experiments where this gene is over expressed. This data was gathered by microscope images.

Figure 2. Live cell imaging of FtsZ being over expressed in NEB turbo cells. Conditions: under a 100x Oil Ph3 Objective Lens and TRANS wavelength in a Zeiss microscope with camera focus. Image is representative of measurements done in triplicate.