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DOI: http://dx.doi.org/10.7551/978-0-262-33027-5-ch031
Pages 147–148
First published 20 July 2015

Ribosome synthesis and construction of a minimal cell using a cell-free expression platform

Filippo Caschera and Michael C. Jewett

Abstract (Excerpt)

The creation of wet artificial life in the laboratory is a non-trivial challenge for biologists, chemists, and computer scientists (1–4). Such a challenge revolves around the modular integration of complex reactions networks to obtain functional biochemical units able of self-replication, self-reproduction, spatial-temporal control and ultimately open-ended evolution, e.g. minimal and artificial cells (1,5–8).

As a step towards building minimal cells, we have developed a cell-free expression system for bacterial ribosome synthesis named iSAT: integrated Synthesis, Assembly and Translation for in vitro construction of Escherichia coli ribosomes (9). The ribosomal RNA, transcribed from its natural operon, self-assembles with ribosomal proteins added to the reaction mixture. Afterwards, in vitro built synthetic ribosomes translate a reporter gene (10,11). Such system is important to design ribosome with new functions, and for the bottom-up construction of a minimal cell.