Team:XMU-China/Safety
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1. The algae part.
HutH, from Pseudomonas putida, can catalyze transformation of L-histidine into trans-urocanate. CBM, modified from Clostridium thermocellum, can target algae cells with its binding ability to cellulose, which is abundant on the cell wall of individual algae cells. Meanwhile, LecA, from Pseudomonas aeruginosa strain PAO1, can bind to galactose in the extracellular polysaccharide (EPS) of algae colonies, enabling SpyCatcher and SpyTag to form a cross-linking network and coagulate the colonies.
All the enzymes were secreted from engineered bacteria through several kinds of signal proteins.
2. The mussel part.
Vibrio natriegens is chosen as the chassis bacteria for its high adaptation to marine environment and advanced background to be operated in genetic engineering.
PPO from Bacillus megaterium is adopted for mussel cleaning. It catalyzes the transformation of dopa into dopaquinone, destroying the adhesion ability of mucin. In addition to remove the attached mussels, TnaA from Escherichia coli strain K12 is also applied to inhibit the synthesis of mucin, preventing the adhesion of mussel.
All the enzymes were displayed on the surface of engineered bacteria through several kinds of anchor proteins, with a plastic adhesion protein LC1KR2 modified from LC1 peptide of Bacillus subtilis to anchor the engineered bacteria on our hardware.
All the proteins are totally harmless to human body. All the plasmids are harmless for human beings and none of our parts would raise any safety issue according to the expertise.
The building of laboratory includes the location of emergency exits, emergency ventilation system, fire-fighting equipment, first aid kit and so on. Every year after recruiting new members, we spend a semester training them in basic laboratory operations and handling safety accidents. Laboratory coat, gloves and glasses are necessary when entering the laboratory. All culture fluid and culture medium with bacteria were processed properly to prevent the potential biological pollution. We handled all the biological materials, wastes and equipment strictly following the BSL-1 requirements, our lab is also equipped with professional operators of dangerous goods and waste biological products, they will disposal all the waste properly. We turn off the water and electricity, close window and door before we leave the lab.
At the same time, we invited laboratory safety engineer, Lina Wu, to conduct safety education for all team members when they officially entered the experimental phase consistently.
