Team:Gaston Day School/Safety

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Safety in iGEM

     We are studying E. coli K12. It is one of the most studied strains of E. coli. It is safe and practical for high school labs, but may cause irritation to skin, eyes, respiratory system, and kidneys. It is dangerous if E. coli K12 is able to transfer its antibiotic resistance to other bacteria, but E. coli K12 itself causes minimal harm to organisms. All bacteria samples and cadmium are maintained safely in a separate lab.
     All team members are required to wash their hands on entrance and exit. E. coli K12 have a very small risk of surviving in the environment so the risk of transfer is already low. All water disposed will go through a wastewater treatment center to make sure all contaminations been removed. All bacteria disposed by the lab will be decontaminated by 10% bleach. All used containers, tubes, and tips will also be decontaminated by 10% bleach or or by treatment for 15 minutes at 15 psi in a pressure cooker. Our materials pose few risks to humans, animals, and plants.
     All students were trained with both safety and lab procedures by our supervisor, Anne Byford. She reviews all our procedures and experiments and worked in a genetics lab for over 10 years. Students were trained to handle hazardous chemicals and bacteria. We dress in proper lab attire with safety glasses and aprons. We also learned how to use the eyewash station and chemical shower. No experiment will proceed without the presence of any well trained members. We teach every member through hand-on experiences and we operate under BioSafety Level 1 because we work in high school laboratory.
     We have also begun design work for a passive killswitch. The kill switch will be arabinose-repressible, incorporating a pBAD promoter for a TetR repressor of a TetR repressible promoter of colicin. When the concentration of arabinose drop, the colicin will no longer be repressed and will kill the cell, preventing any contamination.

Construct 1:

Test the function of the arabinose promoter and determine the most effective concentration of arabinose for induction.

Construct 2:

To be used in combination with construct 3 to test the function of the Tet repressor. the complete test plasmid is construct 4.

Construct 3:

To be used in combination with construct 2 to test the function of the Tet repressor. The complete test plasmid is construct 4.

Construct 4:

Test the function of the arabinose inducible promoter linked to the Tet repressor system. Under arabinose induction, GFP production should fall.

Construct 5:

Intermediate construct containing the lethal gene behind the Tet repressible protein.

Construct 6:

The final kill switch design. Under arabinose induction, the clicin will be suppressed using the Tet repressor. If the level of arabinose falls, as would happen in an accidental release, the Tet repression will stop, causing production of colicin and cell death.