Module 3: Genome Deletion

Overview

Project overview.

Module 3 is the final module of the system. The E.ncapsulator has successfully completed its job of drug production and drug packaging. Now, it needs to be prepared to be converted into a safe pill carrying the drug of interest. This is done by inducing cell death and removing all the potentially harmful genetic material. Removal of genetic material is important to prevent DNA transfer to unintended targets, creating bacteria for instance with higher survivability or more toxicity. This module is a highly reusable system for the removal of genetic material after genes are expressed and the bacteria is no longer needed.

Under control of a thermoinducible promoter, the restriction enzymes DpnII and TaqI are produced. These restriction enzymes cut frequently in the genome, digesting the entirity of the genetic material. Without its genetic material, cell death is also naturally induced. However, restriction enzymes are highly toxic to the cell, where even minute amounts can induce destruction of the genome. Therefore, the native Dam methylation system is used to prevent leaky levels of restriction enzymes from killing the cell prematurely.