Figure. 1 The laser confocal microscopy result of JM109 with no plasmid. There is no GFP gathering in view.

Figure. 2 The laser confocal microscopy results of JM109 with MamC::GFP expression plasmid(the left) and the JM109 strain with both MamC::GFP expression plasmid and artificial gene cluster(the right).

As we see there are obvious GFP signals generally equally distributed in JM109 with MamC::GFP expression plasmid, only a few bacteria contain the GFP gatherings at both ends of cells. And there no GFP signals in middle of the cell. There are GFP gatherings at both ends of cells. Compared with the JM109 with MamC::GFP expression plasmid, the figure(on the right) of JM109 strain with both MamC::GFP expression plasmid and artificial gene cluster shows that there are clear MamC::GFP fluorescence gatherings at both ends and in middle area of the cell, which is different from the results of the other two samples. Additionally, we observed that the shape of the strain with both plasmids has changed. It has become much longer than normal E.coli, as the figure you can see(on the right). And the length of strain with both plasmids is almost three times longer than normal E.coli.

We cannot be completely certain that the protein is actually attached to the membrane as we assumed, because the mechanism of the membrane formation is still not studied thoroughly. We cannot do TEM for the engineered bacteria at the moment due to forces beyond our control. You are welcome to follow us in our future work!