Team:Edinburgh/Safety
Safety
Whole Organisms:
We have only used chassis organisms belonging to Risk group 1 for protein expression, including Bacillus subtilis (168, IIG-Bs27-31, SCK6) and disabled Escherichia coli strains including BL21 (DE3), DH5 alpha and Rosetta in our project. These strains are not expected to survive in the environment in competition with wild type strains.
All strains transformed with plasmids, as well as solids and liquids that were in contact with DNA in general, and constructs carrying the genetic information for antibiotic resistance, are inactivated and disposed of to mitigate the risk of introducing resistance genes to the environment. Therefore, our organisms are not expected to possess any particular environmentally harmful properties.
Parts:
The DNA constructs transformed to the recipient strains will not introduce harmful genes, or gene products that may be of risk to Human Health and Safety. Furthermore, the genes introduced will offer no survival advantage over the systems currently being used by the host. Therefore resulting genetically modified organisms should pose no greater threat to the environment than the unmodified host organisms, and gene transfer will be minimised.
Vectors:
Vectors used in our project are transmissible, but carry no dangerous cargo apart from antibiotic resistance such as kanamycin. These are routinely used in labs for selection purposes and resistance determinants for these antibiotics occur naturally in the environment. Resulting genetically modified organisms should pose no greater threat to health and safety than the unmodified host organisms.
Disposal:
All strains transformed with plasmids, as well as solids and liquids that were in contact with DNA in general, and constructs carrying the genetic information for antibiotic resistance, were inactivated and disposed of to mitigate the risk of introducing resistance genes to the environment. Liquid and solid wastes were disposed of by autoclaving according to University standard protocols (121°C for 15 minutes, and 134°C for 5 minutes respectively).
Safe Lab work:
We did not carry out lab work without the supervision of a responsible scientist, and when we are in the lab we will always wear PPE. Any spillages were decontaminated with 70% v/v ethanol or 1% v/v Virkon. We were instructed in handling and safe disposal of dangerous equipment such as razor blades (used to cut DNA bands out of gels), and supervised during the procedure. Fume hoods were operated under supervision when necessary- for example, when working with trichloroacetic acid and 2-mercaptoethanol during SDS PAGE analysis.