Team:Paris Bettencourt/Safety

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@iGEM_Paris

Summary

In iGEM we take safety seriously. Our project touches sensible topics such as enzyme and human contact so we had a special consideration about safety issues. From strict lab rules to interviews with drycleaners, we tried to measure user's awareness and potential problems in the implementation and safety of our final engineered GMO (genetically modified microorganism).

Safety Training and Lab Rules

Figure 1: Our laboratory facilities at the Cochin institut in Paris.

At the beginning of the competition, all team members received a safety training for working in the Biosafety Level 1 and in the Biosafety level 2 laboratories. This training was conducted by the Biosafety officer for INSERM U1001 (Fig. 1) and it included best laboratory practices as described in the WHO laboratory biosafety manual.
It included rules about protocols in the lab, protective clothing and equipment, decontamination methods and practices, prevention of the transfer of genetic material and microorganisms, handling chemicals, use of machines and emergency procedures. The safety form was submitted to iGEM HQ as well.

Biosafety Level 2

During this project we aimed, amongst other things, to create a database of strains isolated from vineyard soils from all around the world. Since we were handeling samples without knowing the microbes that were present on them, we took special security measurements. To begin with, we worked in a lab with biosafety level 2, in the appropriated hood and wearing the appropriate safety clothing. We also chose to work with a set of media that would be selective for non-pathogenical species, not allowing the potentially dangerous species that were on the samples to grow.

Organisms used

Our sole chassis organism in this project was E. coli. All cloning steps were performed in the DH5alpha strain, and all protein expression steps on the BL21(DE3) strain.
In the making of our database we isolated several different species of Acidovorax, Actinomycetes, Arthrobacter, Bacillus, Brevibacterium, Celullosimicrobium, Corynebacterium, Escherichia, Flavobacterium, Klebsiella, Kocuria, Microbacterium, Micrococcus, Oerskavia, Paenibacillus, Pantoea, Pseudomonas, Rhodococcus, Staphylococcus (non aureus) and Streptomyces. All our samples were sent for sequencing after growing them in selective media and no harmful species for human health was isolated from them.

References

• Public Health Statement for Tetrachloroethylene (PERC) from the Agency for Toxic Substances & Disease Registry of the United States Department of Health and Human Services (2014)