Overview
E. coli is one of the most suitable chassis strains for metabolic engineering modification experiments due to its clear genetic background, as well as its extremely rich library of metabolic components and complete genome editingtechnology.
To ensure the universality of the enzymes we found, we chose the most commonly used engineering strain E. coli to design and construct the cell factories. Safety is undoubtedly the most important in this process, and after analysing the project as a whole, we listed the following.
Experimental Material Safety
Our research initiative involved sampling from diverse soil regions as well as rat gut microbiota; specimens were also obtained from horses, cows goats, camels, and Przewalski's mustangs, feces, human saliva, pseudoplectoides, and poplar tree cavities.
Subsequently, gene sequences extracted from these samples were compiled into a comprehensive metagenomic database before being further processed, into high-quality plasmid vectors for introduction into E.coli BL21,a microorganism on an approved list.
The non-hazardous strains utilized were terminated post-experimentation to prevent any potential escape from containment.
In anticipation of potential field or oceanic deployment for microplastic remediation purposes, the strict evaluation of our bacterial agents will adhere to environmental protection, biosecurity, and agricultural regulations.
Should their release pose any risk of ecological harm or environmental degradation, it would be categorically rejected.
Experimental Process Safety
During the course of the experiment, we may encounter potential threats from two sources: reagents and instruments.
1) Reagent risks: The inducer we are using, IPTG, possesses certain irritant properties. Prolonged skin contact can result in damage, while splashing into the eyes may cause redness and swelling. To mitigate these risks, each team member is equipped with goggles, masks, professional lab gloves, and lab coats. These items are strictly inspected before entry into the laboratory. Additionally, emergency shower facilities are available to ensure the safety of experimenters.
2) Instrument risks: Throughout the experiment, we will utilize equipment such as autoclaves, water baths, ultra-clean workbenches, and centrifuges. The hot steam generated by autoclaves and water baths poses a risk of burns to experimenters; ultraviolet light emitted by ultra-clean worktables can harm their eyes; improper use of centrifuges can lead to more serious consequences. In order to safeguard experimental personnel's well-being, comprehensive training on experimental and instrument operation procedures will be provided prior to commencing experiments. Furthermore, regular inspections of experimental instruments will be conducted by dedicated staff to prevent injuries resulting from equipment malfunction.
Laboratory Safety
In our laboratory, we have implemented a comprehensive safety and hygiene management system following the specifications of standard microbiology laboratories. In order to ensure the strict implementation of this system, we have designated a full-time laboratory manager and safety responsibility person to supervise. Only after completing strict safety training and passing the examination, the personnel are allowed to carry out experimental work.
In the management of chemical reagents, we strictly abide by the relevant regulations of the state and the school to ensure that the procurement, use and storage process is standardized. We also implement the "double lock" management system, strengthen the supervision of the use of reagents, ensure that they are returned in time after use, and record the use in detail to maintain the orderly operation of the laboratory.
