We genetically modified Escherichia coli and Lactococcus lactis in our project. The genes were constructed in E. coli and transformed into Lactococcus lactis using a shuttle vector known as pTRKH2. Both organisms are level 1 (low risk) safety level organisms.
Our safety training was performed by Utah State University Environmental Health and Safety Office and covered chemical hygiene principles, spill prevention, hazardous waste management, and fire safety.
Laws and Regulations that Govern Biosafety in Research Laboratories in Our Country
Information concerning relevant laws and regulations in the United States of America may be found on the U.S. Department of Health and Human Services website at http://www.phe.gov/s3/legal/Pages/laws.aspx.
Risks Posed at the Laboratory Stage and Actions Taken to Reduce Risks
The two main risks posed by our project are exposure to ethidium bromide and releasing genetically modified organisms into the environment. To reduce exposure or release of the genetically modified bacteria we wear rubber gloves and sterilize waste with bleach and/or autoclaving. Biological materials were handled in a biosafety cabinet/laminar flow hood and on open benches.
Ethidium bromide is the main material that we work with that is known to be hazardous, and we wear nitrile rubber gloves when handling this chemical and use the same pipette to transfer it to agarose solutions inside of a chemical hood.
How Our Project Would Be Used in the Real World
Our project would be used in a factory, in a consumer product that ordinary people buy, and in food (cheese cultures).
Future Risks If Our Project Was Fully Developed and Steps That Could Be Taken to Reduce Risks
There is a relatively low level of risk from the bacteria themselves, even though they would be used in food production. The primary concern that may arise is the fact that the bacteria have an antibiotic resistance (erythromycin) in the vector plasmid used to transform our construct into the bacteria. That antibiotic resistance could potentially spread to other bacteria. Since erythromycin is primarily used in treating acne, it's unlikely that the Lactococcus lactis used in cheese culture will come into contact with bacteria causing acne on human skin enough to spread the resistance. Further work that could be done to reduce risk further might be work with a cre/lox system in the plasmid to remove the antibiotic resistance altogether after the plasmid is known to be in the cells.