Results

Plasmid Assembly

The plasmids used as entry vectors and transcriptional units were successfully assembled using golden gate assembly. However, after the downstream testing of transcriptional units used to test the detection mechanism of our genetic circuit, various sequence errors were identified. In order to correct for the mistakes, site-directed mutagenesis was successfully completed to introduce the desired nucleotide changes and restore proper sequence functionality. Please visit Results for more information.

Detection Mechanism Testing

Preliminary trials to determine the functionality of the riboswitch for lead detection was unsuccessful, further experimentation is required. Please visit Results and Future Directions for more information.

Memory System Testing

Using an ortho-Nitrophenyl-β-galactoside (ONPG) assay with yeast expressing the desired serine integrase and reverse-LacZ reporter. Results indicated that the inteagrse is functional in Saccharomyces cerevisiae, however, its efficiency (as determined by relative β-galactosidase activity compared to the positive control) was quite low. Please visit Results and Future Directions for more information.

The filter medium was successfully fabricated and shown to support the growth and adhesion of Saccharomyces cerevisiae in conditions relevant to continuous flow applications. Visual and microscopic analyses confirmed the presence of yeast cells throughout the porous interior of the medium. This verified the structure can sustain biological activity necessary for potential functionality in bioaccumulation-based lead removal applications.

Permeability studies verified that water flows through the rehydrated filter medium at a controllable rate, allowing users to control the contact time between the water and the yeast cells to maximize treatment effectiveness. Additionally, the final 3D printed filter apparatus displayed no leakage or damage, confirming its watertight integrity and compatibility with the filter medium.

The freeze casted filter media were reproducible and demonstrated good structural properties which allow them to retain their structural integrity and shape when subjected to a continuous flow of water.

These results demonstrated the feasibility of the system’s biological, material, and mechanical components, confirming that they can function together as an integrated unit suitable for further development in continuous flow water filtration applications. For detailed experimental methods and results, refer to the Hardware Protocols and Experiments Page.

The BioMap software tool was developed to make computational modelling more accessible to synthetic biologists. The core functionalities including a user-friendly interface, GPT-powered plain language data input capabilities, and interactive simulations were successfully implemented. Visit the Software page to learn more about BioMap and how it supports all synthetic biologists.

Ordinary differential equations (ODEs) for modelling the biological system developed by Wet Lab were formulated. A step-by-step guide to developing ODEs was created and integrated into the BioMap software tool, helping other synthetic biologists generate computational models of their biological systems. Visit the Modelling page to learn more!