1. What is your chassis organism?
Our chassis organism is E.coli . We are using the strain DH5-alpha for cloning and we are going to use the strain BL21(DE3) for expression. These strains are not harmful to humans.
2. What risks does your project pose at the laboratory stage? What actions are you taking to reduce those risks?
The project involves the manipulation of non-pathogenic E.coli (E.coli DH5 alpha and E.coli BL21DE3) so it is currently being developed in safety level 1 in the molecular biology laboratory. In order to respect biosafety rules, we wear lab coats, rubber gloves and sometines cotton masks. All biologial waste has to be treated with chlorine and there is a special section for liquid waste treatment. In molecular Biology organic acids, alkaline solutions and other corrosive reagents will be carefully monitorized and only authorized personal can manipulate it. It is imperative that all the members of the group read the disposal protocols for every kind of solid and liquid waste. We have protocols for acids, alkaline solutions, liquid nitrogen and sterilization of biological material. It is of course forbidden to eat or drink in the laboratory.
3. How would your project be used in the real world?
It could be used in a Factory or by ordinary people as a customer products that they can buy. In a factory, our cells could be used to produce PLA and this PLA could be used to make different customer products like an alternative to conventional plastic products. Industry of packaging, medical devices, fluidized bed reactors and other utilities related to conventional plastics.
4. What risks might your project pose, if it were fully developed into a real product that real people could use? What future work might you do to reduce those risks?
First, people wouldn't use the bacteria as a product but as a way to produce their own PLA. So, before allowing anyone bring at home a bacterial culture we would design protocols to teach people how to safely use the bacteria, how to care them and how to protect themselves and their relatives from them.
Then, people would get the bacteria into a bioreactor that we would have to design too, but which would basically consist in a closed recipient with medium culture, so that any direct contact with people is avoided.
Finally, to prevent bacterial escape and DNA transfer, our bacteria are equipped with a biosafety system, which ensures cell destruction if they leave the biorreactor. This system is arabinose-dependent: the culture medium contains arabinose, which inhibits the apoptosis process. If bacteria escape, the lack of arabinose will trigger the death cell mechanism.
