BYU 2014 Safety

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Safety Overview

Organisms Used

Our team has received safety training according to requirements as set by Brigham Young University's College of Life Sciences. Our training encompassed topics including:OSHA laboratory standard, material data safety sheets, chemical safety including security disposal of hazardous materials, wearing of proper protective clothing, housekeeping, laboratory hazard signs and the NFPA hazard warning system for labels, proper labeling of primary and secondary chemical containers, proper storage of chemicals including chemical compatibility, temperature requirements and secondary containment, disposal of regulated waste, minimizing exposure to hazardous chemicals including engineering controls, administrative controls, and personal protective equipment, proper use of laboratory hoods, chemical spills and proper cleanup. Our university's training requirements may be found here.

The insitutional biosafety committee at Brigham Young University is responsible for biological safety at our university. Our country's regulations that govern biosafety may be found here.

There are specific safety rules and regulations that our team follows to ensure our own safety. These include wearing long pants and closed toe water resistant shoes, and taking proper safety precautions when using chemicals and other biological agents, and no food or drink allowed in the lab. Prior to being able to work in the lab each member is required to complete a laboratory safety course established by the university. Our bio-brick parts do not pose health risks and are only used within the lab. We are working with BSL-1 organisms in our lab, namely E. coli K12 and DH5α and our chassis N. multiformis. We also have used a BSL-2 organism, Psuedomonas aeroginosa PAO1, to provide template DNA for cloning the genes necessary for denitrification. Dr. Julianne Grose, our instructor, has received BYU approval to work with BSL-2 reagents. Our laboratory room has also been approved for BSL-2 lab work, with notification posted outside the room. We have followed current safety measures for disposing of biological samples in biohazard waste as well as cleaning glassware and desktops with bleach/ethanol.

We have each been trained on proper laboratory safety guidelines and follow these rules in our lab. We have chosen to use a BSL1 chassis and are performing experiments withing a BSL-1 strains of E. coli (K12 DH5α). We follow BYU's protocols for the disposal of biohazardous and chemical waste. The parts used in our project have little potential for malicious mis-use.

What about the Environment?

If our project is implemented in water reclamation facilities there is certainly a potential risk for the environment. Our system should be contained within the sewage treatment plant, specifically within the bioreactor, with little to no room for escape of our microbe into the environment during this process. Most water treatment facilities utilize UV exposure or filtration systems to remove microbes from the water before releasing it into the environment, at which stage our designed microbe would be eliminated. To mitigate potential risks of our modified N. multiformis escaping and proliferating in the environment, we plan to knock out the SerA gene, making it an auxotrophic bacterium that would require the high concentrations of serine found inside the bioreactor but absent from other waterways.

• E. coli DH5α This is used for cloning and amplification of plasmids and as a preliminary chassis for our project.
• E. coli S17R This is used for conjugation of our plasmid constructs into N. multiformis.
• N. multiformis ATCC 25196 This is the intended chassis for our project.
• P. aeroginosa PAO1 We are using this genome as a DNA template for the four denitrification genes in our system.
• S. thermophilus LMD9 We are using this genome as a DNA template for the CRISPR genes.