We worked on a fundamental project using E. coli (mostly cell line of 5-alpha and 10-beta) to investigate the effect of protein degradation on the speed of gene expression. Our basic method was co-transforming two plasmids, one that expresses sfGFP with synthetic degradation tags and another expresses mf- Lon protease, to E. Coli cell. The reporter protein concentrations correspond with the degradation strengths of those tags. We used plate reader measurements and time-lapse microscopy to characterize the degradation rates.
Safety and Training in the Lab
We are a Biosafety Level l lab, which means that none of the reagents are known to be pathogenic in healthy humans. We nevertheless followed all BSL-1 precautions, including wearing gloves at all times when working with BSL-1 agents, tightly controlling lab accessibility, appropriately inactivating infectious material through autoclaving and bleaching, banning all food and drink in the lab, and
enforcing rigorous hand washing before leaving the lab to ensure the utmost safety. We received training from multiple sources. Before entering the lab, we received training from our PI in basic chemical safety and biosafety (BSL1). At the beginning of the summer, we also received a refresher course from our PI and reviewed BSL1 protocols extensively. In addition, at the beginning of the summer session, we received two hours of training on all aspects of lab safety from the Director of the Environmental Health and Safety Office, College of William and Mary. All training sessions were documented.
Safety and Ethical Risks
Our project is foundational and therefore we do not have a specific real-world application. For that reason, our project poses no risks in the real world. In terms of ethical risks, these are minimal to non-existent given that we do not plan to use animals or introduce anything into the environment. Since we are doing foundational research, all our experiments will be conducted in the lab with the goal of providing additional knowledge about the role of protein degradation and the speed of reactions in genetic circuits.