Towards a synthetic cell: designing, testing and optimizing functional genomic modules in E. coli

Summary

The research in this thesis was conducted as part of the Dutch Building a Synthetic Cell (BaSyC) consortium, which aims to create a synthetic cell from scratch through a bottom-up approach. The focus of the study was on identifying functional modules that could potentially be used in a synthetic cell and testing them in Escherichia coli as a proof of concept. The functional modules investigated included the ADI pathway for energy production and an RNA polymerase module for transcription. The research also developed a cheap, fast and efficient cloning method in-vivo in E. coli DH10B cells. In addition to testing these functional modules, the research also sought to understand what can be learned from the genome organization of living bacteria when attempting to build a synthetic cell. This information can aid in the creation of a synthetic cell through a bottom-up approach.