Team:Uppsala Universitet/Safety


Lab safety

Sharing is caring when it comes to knowledge in safety. We at Uppsala University care about each-other and that is why we made sure to understand and follow the rules for working in and around the lab. Taking personal responsibility when it comes to following safety rules ensures that no harm will come to ourselves or our teammates. These rules include routines for general building safety, general laboratory safety, biological safety, GMO safety, handling machines and handling liquid nitrogen.

We all participated in the mandatory safety round. First fire extinguishers, first aid kits and showers were introduced and this was followed by an introduction to waste disposal, how to properly mark tubes and how chemicals should be handled. The more dangerous chemicals were handled in a fume hood.

The laboratory work was carried out in a BSL-1 lab (Biosafety level 1) and for personal safety lab coats and protective eyewear were used as well as gloves when needed. Gloves were discarded immediately after use. All organisms that were handled in the lab were classified on the iGEM white-list. The organisms used were Escherichia coli (Strains DH5-Alpha, BL21(DE3)) and Pichia pastoris (Strains X-33, KM71H). Since all the organisms used were on the white-list, no special protective equipment was required when handling samples.

Each member had the opportunity to undertake a guided tour of how waste is processed at the facility where we did our lab work. During the tour, the students were shown where the waste is stored, how to handle containers of waste, the importance of recycling and what to do if a spill occurs. At the end of the tour, each participating member was given a diploma allowing them to handle all waste products that were produced in the lab.

Since often long and stressful days in the lab are a potential hazard on their own, we made sure to communicate any flaw or error in our daily routines at our group meetings and to remind each other of our responsibilities.

Ethics and risks of GMOs

Our goal is to modify the yeast Pichia pastoris so that it expresses enzymes that break down lignin. We have considered the possible danger to trees if our modified organisms were to escape into the natural environment, but judged this risk to be very low. The enzymes produced by our engineered yeast would ultimately be used on dissolved lignin or lignin with a large surface area. They would not work well in the environment. Organisms that break down lignin in trees with the help of secreted proteins already exist in nature, and they are much better adapted to this niche than our engineered yeast cells would be. The lignolytic fungi that break down trees in nature were the very inspiration for this project.

The enzymes we added to Pichia pastoris are naturally occurring and not harmful to the environment. The antibiotic resistance cassette for Zeocin used for selection of our engineered E. coli and P. pastoris cells is standard for this type of research.

In the unlikely event that the engineered yeast escapes from the lab, it would not receive any fitness benefit from its genetically engineered enzymes. The continual production of unneeded resistance proteins and the increased genome size even carry a small fitness penalty. Therefore, it should not be able to compete with natural yeast strains in nature. The same thing applies to the Escherichia coli strains that were used to clone the vector.