Synthetic Biology: (Re)-programming adhesion properties of bacteria - Group 3

The research team explains the biological problem of bacterial biofilm formation and its medical consequences, then introduces the concept of controlling adhesion protein expression to redirect bacterial behavior for beneficial applications like environmental remediation.

Transform naked bacterial cells with plasmids carrying either GFP or mCherry fluorescent genes using mating and electroporation techniques to create distinguishable green and red fluorescent strains.

Perform colony PCR to confirm that the fluorescence genes have integrated correctly into the bacterial genome at the appropriate locations.

Electroporate plasmids containing different adhesion genes and two anchor system types (autotransporters and intimin) into the fluorescently-labeled bacterial strains.

Verify successful electroporation of adhesion plasmids through plasmid extraction or PCR analysis of the transformed bacterial colonies.

Induce expression of the adhesion proteins in the bacteria and verify correct expression and cell surface localization using western blot analysis and fluorescence microscopy.

Present ongoing research needs including intracellular signaling upon adhesion, integration with biosensors, control of adhesin positioning on the membrane, and testing capacity for multiple adhesin types in single cells.