Project design safety
If our project is applied for the real conditions, we can make sure that our system is safe and will not do harm to environment and human. The following is our reason, we use E. coli for production of protein but we won’t use live E. coli but use only proteins in our project so there are no infection problems.
The lab we conducted our experiments is classified as biosafety level. Before starting the iGEM lab work, all members have participated in general lab safety training, and everyone was in full compliance with iGEM' s safety policies.
All the members must follow these rules :
(1)Requiring lab technicians to wear personal protective equipment (PPE), such as protective gloves, laboratory coats.
(2)All laboratory workers should undergo simple first aid training.
(3)Dispose of any hazardous material as toxic waste
(4)Keep the workplace tidy. A tidy laboratory avoids accidents to everyone.
Project Specific Laboratory Safety
When we are testing the capture efficiency of activated carbon in water that contains BPA and NP, we do take the risk of exposure to EDCs. To avoid the danger of EDCs, there are some precautions and notices:
(1)Wearing personal protective equipment including safety glasses, protective gloves, full body suits, respirators and so on.
(2)Throwing gloves into biological waste treatment barrels and wash hands thoroughly after the experiment.
(3)Pouring wastewater containing BPA and NP into chemical waste storage bucket.
Biosafety is the top priorities to be considered in the iGEM competition. By the definition from WHO(World Health Organization), biosafety is ”the safe working practices associated with the handling of biological materials, particularly infectious agents. It addresses principles, technologies, and practices that are implemented to prevent the unintentional exposure to pathogens and toxins or their accidental release.”
Based on the above concerns, we decided to choose E. coli, DH5α, BL21and MG1655, a nonpathogenic and relatively common bacteria, as our chassis. Moreover, the parts we used are safe to our environment and health:
(1) Part:BBa_K2354010 :monobody-GBP HS
BBa_K2354010 is a part which can make E. coli produce the protein with the function of monobody and gold binding protein. The function of monobody is to bind to ER- alpha/EDCs complex and the function of the gold-binding protein is to bind to the gold surface. As a result, this fused protein can’t make any negative effects on health or environment.
(2) Part:BBa_K2354011 :INP-ER-alpha HS
BBa_K2354011 can make E. coli express ER-alpha on the membrane and ER-alpha can bind to estrogen activity molecule. This ER-alpha expressed E. coli just have an additional function to capture EDCs in water and it is harmless to health or environment. Moreover, we froze E. coli to lower its activity so we make sure its safety further.
(3) Part:BBa_K2354012 :HRP-his tag
BBa_K2354012 can make E. coli produce horseradish peroxidase and the function of this protein is to degrade EDCs with phenolic groups in the water. Horseradish peroxidase is a kind of peroxidase which uses heme as the cofactor and it doesn’t have any toxin structure or molecule. Therefore, this part is harmless to health or environment as well.