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Cloning whole bacterial genomes in yeast

2010; Oxford University Press; Volume: 38; Issue: 8 Linguagem: Inglês

10.1093/nar/gkq119

1362-4962

Gwynedd A. Benders, Vladimir N. Noskov, Evgeniya A. Denisova, Carole Lartigue, Daniel G. Gibson, Nacyra Assad-Garcia, Ray-Yuan Chuang, William Carrera, Monzia Moodie, Mikkel A. Algire, Quang N. Phan, Nina Alperovich, Sanjay Vashee, Chuck Merryman, J. Craig Venter, Hamilton O. Smith, John I. Glass, Clyde A. Hutchison,

Plant Virus Research Studies

Most microbes have not been cultured, and many of those that are cultivatable are difficult, dangerous or expensive to propagate or are genetically intractable. Routine cloning of large genome fractions or whole genomes from these organisms would significantly enhance their discovery and genetic and functional characterization. Here we report the cloning of whole bacterial genomes in the yeast Saccharomyces cerevisiae as single-DNA molecules. We cloned the genomes of Mycoplasma genitalium (0.6 Mb), M. pneumoniae (0.8 Mb) and M. mycoides subspecies capri (1.1 Mb) as yeast circular centromeric plasmids. These genomes appear to be stably maintained in a host that has efficient, well-established methods for DNA manipulation.