Our university takes the safety of the students very seriously. It provided all the team members with appropriate safety training prior to the start of the project. We attended a workshop on biosafety where our university safety officer, Dr Stuart Thompson spoke to us regarding biosafety and how to be safe in the laboratories as well as making us aware of the university policies. He provided us with 15 basic rules to follow in the laboratory (end of the page). To ensure that the project is being done with all safety procedures considered, we followed all the requirements given by the university as well as the iGEM safety policies.
The team used personal protective equipment (PPE) including lab coats and gloves at all time, googles where used when required. We also made sure to wear appropriate clothing, and hair tied back while being in the laboratories to ensure reduced contact with biological material as wells as reducing the possibility of contamination.
Two members of the team participated in the University’s genetic modification safety committee meeting, where the committee went over their GMM form and made sure that all the safety requirements are being considered and there is no biohazard.
The aim of the project is to tackle the issue of polystyrene. We genetically engineered an E. coli bacterial strain to carry commercially synthesised genes of the tod operon found in Pseudomonas putida F1 strain that allows E. coli to grow on styrene as a carbon source. The tod operon allows complete degradation of toluene by the P. putida. We repurposed this pathway for the biodegradation of styrene as styrene is a similar molecule to toluene.
The synthesized genes are from the tod operon pathway, which is involved in toluene metabolism and has no known threat to mammalian cells. The plasmid vectors are all based upon pUC18 or pUC19 and are therefore non-mobilisable.
Genes to be synthesized commercially
todE - to convert 3-Vinylcatechol Acrylonitrile to 2-Hydroxy-6-Oxocta-2,4,7-trienoate
todF - allows the organism to live on styrene
todX - allows styrene to be transported within the organism/bacteria to be degraded
1. At all times you must wear correct protective clothing- a laboratory coat which must be correctly fastened at all times. The School of Biosciences provides laboratory coats for students. These MUST be put on a coat hanger and returned to the rack after use, and NOT thrown on the bench or floor. Depending on the experimental design, additional protective wear should be worn (e.g. gloves, masks, safety spectacles, visors) as instructed by the member of staff in charge.
2. You must not eat, drink, smoke or apply cosmetics in the laboratory, on any laboratory floors, on stairs, or in lifts.
3. You must not suck or bite pencils or pens. You must not wear sandals, cumbersome jewellery or hats.
4. You must not lick labels prior to sticking them to apparatus (use tap water, self-adhesive labels or marker pen).
5. Avoid touching your face, hair, eyes, mouth, etc. whilst in the laboratory. Long hair must be tied back.
6. You must keep your available bench space clear, clean, tidy and free of inessential clutter, e.g. books.
7. You must not remove any materials from the laboratory, e.g. microbial cultures, without the permission of the lecturer in charge.
8. Manipulations by loop or pipette should be performed in a manner to minimise the production of aerosols.
9. Pipetting by the mouth of any material is forbidden. You must always use the teats, syringes, and pipette-fillers provided.
10. All manipulations should be performed aseptically, using plugged pipettes, and the contaminated pipettes disposed of in the containers indicated by the lecturer in charge.
11. Contaminated glassware, plasticware, microscope slides and discarded Petri dishes etc., must be placed in the receptacles indicated by the lecturer in charge.
12. It should be recognised that certain procedures or equipment produce aerosols of contaminated material, e.g. the breaking of any fluid film, centrifugation and the agitation of fluids in shaking or orbital incubators.
13. Report all personal accidents, minor cuts and abrasions, breakages and spillages of cultures and reagents to the lecturer in charge. Cuts and abrasions must be protected by an adequate waterproof dressing.
14. If instructed, before leaving your bench, swab the area down with an appropriate disinfectant.
15. Before leaving the laboratory, return personal protective clothing, hang up your lab coat correctly, and wash your hands with soap (preferably germicidal). Before leaving the laboratory, return personal protective clothing, hang up your lab coat correctly, and wash your hands with soap (preferably germicidal).