Project
Our lab works under Biosafety Level 1. We chose this because our team operates in an active classroom, meaning that many safety measures must be in place in order to protect student safety. For our project, we chose the bacteria E. coli K-12 because it is one of the least harmful bacteria available and also has the least amount of side effects. Each iGEM member learns safety and lab procedures and how to work with dangerous chemicals and bacteria from Anne Byford, our supervisor. She also teaches us how to use the eyewash station and chemical shower. Our supervisor answers all questions and looks at our experiments and procedures. Before leaving the lab, every member washes their hands with warm soap and water. Food and drink are not allowed inside the lab. New iGEM members are closely supervised by either Anne Byford or one of the leaders until they have correctly learned the procedures. We disinfect all containers, tubes, and tips with either 10% bleach or 15 minutes in a pressure cooker at 15 psi. We also disinfect every type of bacteria with 10% bleach before we dispose of them. When not in use, the laboratory stays locked. Tips, gloves, and centrifuge tubes are properly disposed of. To keep the classroom safe, we bleach all tables with 10% bleach at the end of each day.
Kill Switch
Two years ago, we started work on a kill switch in case E. coli escaped the lab. It uses an arabinose induced promoter which, when turned off, will cause the cell to create the toxin colicin, thus causing the cell to kill itself shortly afterward. We chose arabinose because it does not appear in nature and can be controlled relatively easily in the lab. Arabinose acts as an environmental cue, and when it leaves the lab, it should kill itself more quickly than other kill switches like those activated by light. Although the project is not complete, we plan to finish it in the future.
Side Effect
As a part of our current project, we may have to grow our E. coli on glucose to help reduce the metabolic load. This would have two main side effects. It would increase the bacteria’s isobutanol production, and they would also only be able to grow in a glucose rich environment. Such an environment could not exist in nature, acting as a form of killswitch should the bacteria leave the lab. A glucose rich environment would allow the cells to produce large amounts of isobutanol without running into the issue of not having enough glucose to live.
Isopropanol Resistance
Our team has deliberately put off serious testing regarding isopropanol because we do not want to create isopropanol resistant bacteria. Creating such bacteria could produce disastrous results as it is the active ingredient in hand sanitizers. Though we wash our hands thoroughly after working with E. coli, we cannot run the risk of having bacteria that resists hand sanitizer leaving the lab because of its frequent use outside of the lab. The main problem is that a large number of people think that hand sanitizer works just as well as soap and water, as well as the fact that most hand sanitizers are advertized to kill almost all bacteria in seconds As a result, some people will use hand sanitizer in lieu of washing their hands. Hand sanitizer is also used largely in hospitals, meaning that isopropanol resistant bacteria would have a higher chance of being transmitted from patient to patient should such a resistance be created.
