Team:JiangnanU China/Safety

JiangNan

1. Safe Project Design

1.1 Chassis microorganism


The E. coli BL21 used in our experiment is one of the most commonly used engineered bacteria in the laboratory. It is on the white list given by the iGEM committee. Even if they flee, they will not survive,with no adverse impact on the environment or the community.

1.2 Phage


The bacteriophage used in the experiment, T4 bacteriophage, is on the white list given by the iGEM committee. It is very safe and does not cause any harm to human beings. We have been very careful during the course of the experiment to try to reduce the risk of phages infecting other normal E. coli bacteria in the laboratory.

1.3 Genetic Components


The inducible promoters, fluorescent protein gene, resistant protein gene, toxic protein gene and other gene components used in gene circuits come from E. coli BL21 genome or the existing gene component library, which will not cause safety problems.

1.4 Kill Switch


The bacteriophage resistant strain we constructed may be used in industrial fermentation in the future, which will significantly improve the resistance of industrial fermentation strains against typical bacteriophages. If the strain is misused, it is likely to promote the evolution of typical phages which are common in our lives. Once the evolution is unable to prevent the stage, will bring huge losses to the fermentation industry. At the same time, the bacteriophage resistant components we used combined with the cold-resistant, heat-resistant and hyperosmotic components built by the previous team, it is possible to create superbugs. Therefore, we introduced the toxic protein used by the previous team as a suicide device to control the life activities of bacteria to ensure the safety of the project.

2. Safe Lab

2.1 Standard Experimental Operation


Before carrying out the experiment, all the experimental personnel have carried out strict experimental safety training and assessment. Only after passing the assessment can they use the laboratory instruments. During molecular experiments, we need to perform electrophoretic validation of DNA and protein expression, which involves exposure to low toxic chemicals such as GlodView, MBA, and SDS. Therefore, during agarose gel electrophoresis and sds-page electrophoresis, we all wear rubber gloves, respirators and other protective tools as required, and strictly control the use and discharge of reagents.
When conducting bacteriophage-related experiments, we immediately sterilized the used experimental tools with high temperature, high pressure and humidity to prevent the escape of bacteriophages and adverse effects on other E. coli in the laboratory. After the experiments, we will conduct high temperature, high pressure, humidity and heat sterilization treatment for all the experimental waste that needs to be discharged, and disinfect and tidy the used workbench to prevent bacteria or phages from escaping.

2.2 Professional Safety Supervisors


Rao Zhiming, professor of biotechnology at Jiangnan University, is responsible for managing the safety issues of our entire project and serves as a safety supervisor. He has been engaged in the fermentation industry for a long time and is responsible for laboratory safety. He is very familiar with the strains used in our project and the operation in the experiment process, and gives us a lot of practical guidance for the whole project.

2.3 Strict Laboratory Management


All experiments in the project were carried out in strict accordance with laboratory rules. The experimental consumables shall be purchased by specially-assigned persons, and relevant equipment such as PCR instrument and ultra-clean table shall be managed by designated persons, and registration shall be conducted for each use.

2.4 Safety Lecture


Before the experiments, we invited several teachers to give us safety lectures, with the purpose of improving the safety awareness of the experiment and standardizing the operation of the experiments.

3. No Release Policy

As a responsible iGEM team, we strictly abide by the no release policy. The wet experiment project is only carried out in the school laboratory and is limited to the experimental stage. No GMOs or GMOs products are released or deployed outside the laboratory.

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