Grand scale genome manipulation via chromosome swapping in Escherichia coli programmed by three one megabase chromosomes

Tatsuya Yoneji, Hironobu Fujita, Takahito Mukai, and Masayuki Su’etsugu

Nucleic Acids Research, gkab298, doi: 10.1093/nar/gkab298

Recent advances in synthetic biology have made it possible to create a bacterium programmed by a synthetic genome. Past work demonstrates synthesis of a whole bacterial genome in yeast, and subsequent genome transfer into bacterial cells. However, the genome transfer has only worked for a small closely related group of Mycoplasma species whose chromosome is relatively small (roughly 1-Mb). To enable the genome transfer approach in E. coli, whose chromosome size is originally 4.6-Mb, the authors developed a E. coli strain programed by three 1-Mb split-chromosomes. Each split-chromosome was purified as supercoiled DNA molecules. Two of these split-chromosomes were then individually transferred into E. coli cells by electroporation. This method could accelerate the booting up of synthetic genomes in commonly used bacteria.

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