The goal of our project is to engineer a set of hydrogel-based whole cell biosensors by utilizing the LasI-LasR quorum sensing system originally found in P. aeruginosa. This whole cell biosensor contains a lacZ gene as the reporter gene, encoding β-galactosidase. The detection and quantification of blue β-galactosidase activity after incubation in an 5-bromo-4-chloro-3-indolyl-b-galactoside (X-gal) solution act as a visible readout for the amount of quorum sensing signalling molecules, N-acylhomoserine lactones (AHLs) which ultimately used for rapid, sensitive and quantitative detection of microplastics.
Our team selects non-pathogenic classis for usage. They are safe strains of Escherichia coli called DH5α and BL21 to transform recombinant plasmids - pSB1C3 which carries the protein-coding sequences to produce the proteins. Both of the E. coli strains that we choose have no harm on humans nor the natural environment. There is no genetically engineered bacteria released into the environment.
All organisms involved in our experiments pose no serious risks even if escaped from the lab. No hazardous chemicals are used in our experiment. We utilized antiseptic techniques and used Bunsen burner to maintain sterility of the workplace and experimental apparatuses. Our project uses a non-pathogen and not use animals. Our lab is of low risk, and it has safety systems in place, for example eyewash bottle and fire extinguisher. The waste that is generated throughout our project will be disposed of in the biohazard bin.
Our team's PI is an experienced biology teacher and is fully capable of overseeing student members and maintaining lab safety. Also, all student members in our team will only do experiments in the lab under the supervision of our team's PI and a qualified lab technician. Our team members have also received formal safety and security training.