Determination of E. Coli Cellular Factors That Contribute to Survival Upon Exposure to the Alkylating Agent Benzyl Bromide

Alkylating agents have the ability to add alkyl groups onto DNA at various positions. There are many of these agents present in our environment that can ultimately lead to damaged DNA and mutations. While some mutations are beneficial many of the ones caused by these chemicals can have detrimental effects. Benzyl bromide is a known alkylating agent and can be a byproduct of industrial processes and then released into the environment. The goal of my work is to identify genes responsible for cellular responses to DNA damage caused by benzyl bromide using a library of E. coli knockout strains. E. coli knockout strains are being created using bacteriophage‐mediated transduction and the identities confirmed by colony PCR and DNA sequencing. The genes deleted in the knockout strains are those that encode various proteins that play a role in some aspect of DNA repair. While carrying out these experiments the knockouts in comparison to the wild type E. coli are expected to have a lower survival rate. After optimizing the assay conditions, we are determining the survival of strains in our library of now more than 30 knockout strains. Insight to how these E. coli strains react when exposed to this alkylating agent will give us more information on how these genes affect DNA repair and allow further studies to be done. We will be able to determine which genes have a more significant role in comparison to others and focus on other causes and possibly ways to reduce its effects. Support or Funding Information Supported by American Cancer Society (RSG‐12‐161‐01‐DMC) and NIEHS ROUTES program.