Part:BBa_K302012:Experience

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Applications of BBa_K302012

The filamentous cell part is repressed by LacI. The LacI repression is removed by induction with IPTG. Without LacI repression a higher proportion of the cells are filamentous. The filamentous cell phenotype is achieved by overexpressing yneA which is an indirect inhibitor of cell division.

We characterised the part first without, and then with, LacI repression (using the integration vector pMutin4 to integrate lacI into the Bacillus subtilis 168 chromosome.

Graph1:

Graph 1 shows that overexpression (no Lac repression) of the yneA gene (ΔamyE:pSpac(hy)-oid::yneA) leads to a longer cell length compared with our control Bacillus subtilis 168.

See below: Bacillus subtilis 168 cells (left),Bacillus subtilis expressing yneA(centre) and Bacillus subtilis overexpressing yneA(right)

Graph2:

Graph 2 shows the percentage of cells at different lengths(μm)uninduced

See below: Bacillus subtilis 168 cells (left) and non-induced cells(right)

Graph3:

Graph 3 shows the percentage of cells at different lengths(μm)induced at 0.2mM IPTG

See below: Bacillus subtilis 168 cells (left) and cells induced at 0.2mM IPTG(right)

Graph4:

Graph 4 shows the percentage of cells at different lengths(μm)induced at 1mM IPTG

See below: Bacillus subtilis 168 cells (left) and cells induced at 1mM IPTG(right)

Graphs 2,3 and 4 show a greater proportion of cells at a higher concentration of IPTG(1mM IPTG), compared with Bacillus subtilis 168 our control population.

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