Safety
Project Design
E. coli JM109 strain that we are using is derived from E. coli K-12 strain which is in the Risk Group (RG1). RG1 organisms are not associated with disease in healthy adult humans with not carrying any active virulence factor, so we believe that it will pose only a minimal health risk if it escapes the lab.
Our bacteria will be engineered to detect carbon monoxide. We intend to introduce the carbon monoxide sensor protein CooA and the photoprotein luciferase into E. coli. These genes expressed are not known to cause any harm to humans. Our laboratory activities will be carried out using standard lab protocols.
Training
All members attended “Genetic Recombination Experiment Education and Training” before starting lab works and learned the laws and risks related to biocontainment.
Fig. 1 “Genetic Recombination Experiment Education and Training”
Lab Work
Our University Rules We adhere to biosafety and biosecurity rules and standard operating procedures provided by our university, which are compliant with national biosafety and biosecurity laws and regulations. Here we provide a translation of our university’s policy for genetic recombination and the link to its webpage (Japanese).
Kyushu University’s Procedures for Genetic Recombination Experiments:
1. The principal investigator of the experiment should submit the experiment plan to the administrative staff of his/her department after preparing it.
・This will be confirmed by the Chief Safety Officer and the application will be submitted to the President.
2. If you are a member of a research team, you will be required to submit an application for approval of your experiment. The results of the review will be notified to the principal investigator through the department head.
3. Please start the experiment after receiving the approval notice.
* Genetic modification experimental facilities other than P1 level require approval by a separate committee. For more information on how to obtain approval, please refer to "Flow of Procedures for Prevention of Proliferation of Genetic Modification Experimental Facilities".
* In order to apply for approval of a genetic modification experiment, you must have attended the "Education and Training for Genetic Modification Experiment Workers".
URL: http://ura.kyushu-u.ac.jp/dna/
Our Lab Practice
・Pipette tips and Petri dishes with E. coli on them were placed in autoclave bags and sterilized before disposal. Also, when using the autoclave, we took care never to open it until the temperature inside was below 80 ℃.
・Hand sanitization was performed frequently.
・Reagents that pose a risk to health, such as Midori Green, were used with gloves.
Safety for Our Project
Our project uses CO, which is a toxic, flammable, high-pressure gas, so you need to be very careful when using it. In addition, proper storage and management are required to prevent misuse for crimes.
Proper use
We made a chemical risk assessment before using CO to consider the risks associated with using it and the measures to reduce that risk. I think we were able to significantly reduce the risk of inhaling CO by using it in the draft.
Proper management
CO was stored on a locked shelf, distinguishing it from other chemicals. In addition, Kyushu University has a chemical substance management support system "IASO", which thoroughly grasps inventory and usage fees.