Module 6: Containment

Description | Design | Construction | Characterisation | Modelling | Results | Conclusions

Characterisation

"The days of the head-shaking dismissal of the questions concerning risks, and the easy answers referring to the vast experience from the laboratory experiments are over; they have been replaced by scientific investigations of the questions and increased activity in the field of ecology. This is an interesting example of scientific choice being influenced by events in the rest of the society." (Molin et al. 1993)

In order to determine the activity of the periplasmic nuclease, we will be utilising DNAse agar. This agar contains DNA fragments that would be cleaved by the nuclease, leaving the regions surrounding the colonies free of genetic material. The agar plates can then be stained with a DNA indicator such as Hydrochloric Acid or Toludine Blue, allowing the DNA voids in the agar to be visible.

We further characterise the nuclease Biobrick BBa_K729004/BBa_K729004, showing that material from cells containing our Biosafety BioBrick is not able to transform commercial competent cells. We suggest periplasmic nuclease expression provides a promising strategy for preventing transfer of genetically modified DNA. This is particularly valuable for the application of synthetic biology in environmental contexts.

For the toxin/anti-toxin pairs characterisation, we will be transforming our toxic plasmids into cell both with and without the genomic anti-toxin genes. By comparing the ratio of cell viability in each case, we will be able to identify how effective our containment system is.