Our submitted BioBricks

<groupparts>iGEM015 Berlin</groupparts>

Functionality of our BioBrick Prom110_FliC_MCS (BBa_K1617003)

A = MG1655 z1 ΔFliC B = MG1655 z1 wt C = Prom110_FliC_MCS (BBa_K1617003)

To proof and validate that our designed construct works as expected we performed a motility assay. We have designed the flagelline expressing gene FliC with integrated multiple cloning site and cloned it into PSB1C3 with the constitutive promotor BBa_J23110. The figure shows the comparison regarding the motility between our construct (right) and a negative (left) and positive (middle) control. Since our construct was transformed into a MG1655 z1 ΔFliC strain we used the highly motile wildtype MG1655 z1 as a positive control and as negative control the FliC knockout strain MG1655 z1 ΔFliC.

The pictures of negative control and construct were taken after 14 hours of incubation at 37 degrees. The one of the positive control was taken after 5 hours since after 14 hours this control was not evaluable anymore due to the fact that the plate was completely overgrown. Our motility assay reveals that our designed construct which was transformed into a flagelline gene deficient strain is able to successfully express flagellines which are folded correctly and integrated onto the surface of the cells. Compared to the highly motile wildtype strain the motility of our construct is less pronounced but in contrast to the negative control there is a significant difference in the motility visible Our BioBrick enables the FliC deficient strain to be motile again.