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Cloning whole bacterial genomes in yeast.

ABSTRACT: 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.

SUBMITTER: Benders GA

PROVIDER: S-EPMC2860123 | biostudies-literature | 2010 May

REPOSITORIES: biostudies-literature

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Cloning whole bacterial genomes in yeast.

Benders Gwynedd A GA Noskov Vladimir N VN Denisova Evgeniya A EA Lartigue Carole C Gibson Daniel G DG Assad-Garcia Nacyra N Chuang Ray-Yuan RY Carrera William W Moodie Monzia M Algire Mikkel A MA Phan Quang Q Alperovich Nina N Vashee Sanjay S Merryman Chuck C Venter J Craig JC Smith Hamilton O HO Glass John I JI Hutchison Clyde A CA

Nucleic acids research 20100307 8

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 M ...[more]

PMID: 20211840

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