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 BSL1 organisms in our lab namely E.Coli K12 and DH5alpha, with N. Multiformis as our chassis. We also have used BSL2 organism, Psuedomonas Aeroginosa, to use as template in amplification of genes 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 BSL1 strains of E.Coli (K12, DH5alpha). 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.

If our project were to be commercially available for the improvement of water reclamation processes there is potential risk for the environment. Primarily our system should be contained withing the sewage treatment plant and specifically within the bioreactor portion of this process. There should be little to no room for escape of our microbe into the environment during this process. However, if it were to escape into the environment it could pose ecological concerns where our engineered N.Mulitformis could outcompete native soil bacterium.

To mitigate potential risks associated with our project we plan to knock out the SerA gene from N. Multiformis making it an auxotrophic bacterium. Where only the sewage treatment facility, where our bacterium would be put in use, would have the serine necessary for survival.

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Your Lab

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• E.coli DH5α used for cloning and amplification of plasmids. A preliminary chassis for our project.
• E.coli S17RThis is to use for conjugation of our plasmid constructs into N.multiformis
• N.multiformisATCC 25196This is the intended chassis for our project.
• P.aeroginosaPAO1 We are using this genome as a DNA template for the four denitrification genes in our system.
• S.thermophilusLMD9We are using this genome as template for the CRISPR genes.