Abstract

Cell-free expression systems are widely used to synthesize proteins for subsequent further characterization, to manufacture potentially useful commercial end products, and to construct cellular mimics in the laboratory. The first part of the thesis explores the feasibility of preparing two of the commercially available and widely used E. coli-based cell-free expression systems: the PURE System and the S30 Bacterial Extract. The second part focuses on the characterization of in vitro transcription and translation. The third part of the thesis features an example of an application of S30 Bacterial Extract cell-free expression systems i.e. the building of cell-like structures that can work together with engineered bacteria to achieve a predetermined task. Finally, the construction of a microfluidic dialysis device compatible with cell-free synthetic biology projects is presented.