Safety has always been a major concern through our project, both in the experiments design and operations. We used two kinds of bacterium, one is Escherichia coli JM110 and the other is Bacillus amyloliquefaciens LL3, which belong to Enterobacteriaceae and Bacillaceae, separately. According to the official iGEM guidelines of Risk Groups, both E. coli and Bacillus amyloliquefaciens are categorized as Risk Group 1 organisms that “do not cause disease in healthy adult humans” when used properly. The safety level of our lab is P2, which is safe enough to perform all our experiments. We highly valued the individual safety, environmental safety and biological safety.
1. Individual Safety and Environmental Safety
All the iGEMers in our project have taken safety training courses in our department experiment center, which include fire-fighting strategies, disposition of biochemical materials, proper usages of microwave oven and so on.
All members are required to wear nitrile gloves, masks, closed shoes in the lab to prevent potential threats of personal health. What’s more, we separate resting areas from experiment areas and confine eating and drinking in resting areas.
Biological materials were not allowed to be taken outside the laboratory without sterilization. After sterilization, these wastes are collected and decontaminated by the specialists. Regular cleaning of our laboratory trash was arranged every day to maintain a clean and safe environment.
2. Biological Safety Concern Based on the Project
With the advent of synthetic biology, more and more genetically modified microorganisms are used for biomedical, industrial and environmental applications. Both Bacillus amyloliquefaciens and E. coli we use are safety level 1 organism. Besides, we separated our Bacillus amyloliquefaciens LL3 from Chinese fermented food. Still, we take steps to prevent these genetically engineered organisms' diffusions to the utmost.
Editing the genome of Bacillus amyloliquefaciens LL3 for aspB and lysC gene to construct a kind of Asp auxotroph(ΔaspBΔlysC).
Add mf-lon ssrA tag into essential genes by genome editing. We can use biocontainment signal to control mf-Lon protease expression and block essential genes expression to kill bacterium upon loss of the biocontainment signal.
Due to the time limitation and the difficulty of genome editing, we only succeeded in step one in the original strain.
3. Safe Shipping
We sent our BioBricks through the standard shipping process required by iGEM headquarter.