Creating Bacterial Strains from Genomes That Have Been Cloned and Engineered in Yeast

Abstract

Character Transplant When engineering bacteria, it can be advantageous to propagate the genomes in yeast. However, to be truly useful, one must be able to transplant the bacterial chromosome from yeast back into a recipient bacterial cell. But because yeast does not contain restriction-modification systems, such transplantation poses problems not encountered in transplantation from one bacterial cell to another. Bacterial genomes isolated after growth in yeast are likely to be susceptible to the restriction-modification system(s) of the recipient cell, as well as their own. Lartigue et al. (p. 1693 , published online 20 August) describe multiple steps, including in vitro DNA methylation, developed to overcome such barriers. A Mycoplasma mycoides large-colony genome was propagated in yeast as a centromeric plasmid, engineered via yeast genetic systems, and, after specific methylation, transplanted into M. capricolum to produce a bacterial cell with the genotype and phenotype of the altered M. mycoides large-colony genome.