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Before being allowed into the laboratory, all wet lab team members who will be operating experiments in the laboratory need to complete a compulsory laboratory induction session for PC2 BioSafety lab. It contains the following modules:

• Biological safety course
• Ergonomic and manual tasks
• Fume cupboards and safety
• Gene tech regulation at UNSW
• Green lab compliance
• Hazardous substances online
• Health and safety awareness
• Laboratory safety awareness

In addition, all laboratory members were trained in the use of special equipment and had the ability to perform experiments in a safe environment. Personal Protective Equipment (PPE) requirements were strictly followed where lab gowns, gloves, and safety goggles were worn at all times to avoid contact with potentially hazardous compounds. Chemicals were handled with caution and were labeled and stored appropriately. Wastes were disposed of according to the waste disposal protocols.

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In our project, we needed to verify the effect of synthetic peptides on Puccinia spp; a fungus, but for the safety of our experimenters, we used the bacteria Escherichia coli instead of fungus to observe the effect of synthetic peptides on Puccinia spp.’s protein.

The aim of our project is to kill Puccinia spp, the cause of the cereal rust disease. To achieve this, we have selected proteins that play an important role in the metabolism of Puccinia spp. as target proteins and designed peptides that are able to bind to them. Binding the target protein with the designed protein would lead to protein loss of function and thus fungal death. However, according to the conditions of the PBL2 experiment, we cannot use the fungi in the laboratory. This is because the fungus is potentially harmful and pathogenic. Even if it doesn't pose a significant risk to our experimenters, it could pose a risk of infection to plants it may come in contact with if contamination occurs. In contrast, E. coli are bacteria that can be found in many environments and majority of them are harmless or do not pose a significant risk to our experimenters. Thus, we used E. coli to express the target protein in the experiment and to verify the effect of the engineered peptide on the target protein. We used the protein from the Puccinia spp. in our experiments rather than the fungus itself to avoid possible contamination and harm.

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As we work with GMOs in our project, we will incorporate a toxin/antitoxin kill switch system into our design to ensure that GMOs do not spread in the environment where crop rust is not present.

More information can be found in the safety concern module of the proposed implementation.

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